QUADRA 4 Manual Rev-E2

Transcription

®
QUADRA 4
OPERATOR’S
MANUAL
TOMTEC®
REV: E2 – March 2015
1000 SHERMAN AVE., HAMDEN, CT 06514
PHONE 203-281-6790 FAX 203-248-5724
Toll Free (877) 866-8323
http://www.tomtec.com
TABLE OF CONTENTS
Preface ....................................................................................................................9
Unpacking Instructions...........................................................................................10
Locating the Quadra 4 ...........................................................................................10
Section I
General Information .............................................................................................11
Specifications & Technical Information ..................................................................12
Warnings, Cautions, Notes, and Symbols..............................................................13
Introduction & Login Instructions............................................................................14
Quadra 4 – Pictorials .............................................................................................15
Quadra 4 Front View ..................................................................................16
Quadra 4, Side Views..................................................................................17
Quadra 4 with Stackers, Rear View.............................................................17
Quadra 4, without Stackers, Rear View.......................................................18
Quadra 4, Access Panel..............................................................................19
Compressor & Pneumatics..........................................................................20
Section II ...............................................................................................................21
Operational Functionality
Pipetting System ....................................................................................................21
Quadra 4 General Information ...............................................................................22
Quadra 4 Product Line ...........................................................................................23
Section III ..............................................................................................................25
Quadra Command System v4 Software
1.0 Introduction ......................................................................................................26
1.1 What’s New ...........................................................................................26
1.1.1 Tip Load Counter …………………………………………………………27
1.1.2 PSID2 Mirror Rack Checking …………………………………………. 28
1.2 Quadra 4 DMV Head.............................................................................29
1.3 Getting Started ......................................................................................29
2.0 Main Screen.....................................................................................................31
2.1 Labware Components Box ....................................................................31
2.2 Program Operations Box.......................................................................32
2.3 Program Data Box.................................................................................32
2.4 Shuttle Position Data.............................................................................32
3.0 Program Operations.........................................................................................34
3.1 Arm Plate Transfer Operation ...............................................................34
3.2 Aspirate .................................................................................................35
3.3 AutoTrap Operations .............................................................................36
3.4 Cell Wash..............................................................................................37
Quadra 4 Operator Manual
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TABLE OF CONTENTS
3.5 Delay .....................................................................................................39
3.6 De-Lidder Operation..............................................................................39
3.7 Dispense ...............................................................................................40
Calibration Table
Dispense Speed
Dispense Height
Blow Out Volume
Touch Off
Dip Tips
3.8 Empty Sample.......................................................................................42
3.9 GOSUB Program ..................................................................................44
3.10 GOTO Program ...................................................................................45
3.11 Load Tips ............................................................................................46
3.12 Lockout Shuttle Position......................................................................47
3.13 Loop ....................................................................................................48
Infeed From Stacker(s)
Outfeed To Stacker(s)
Number of Cycles
Disable Lid Capability
3.14 Magnetic Nest .....................................................................................49
Select Device
Action
Toggle Action
Lockout Position
Seconds
Shuttle Position
3.15 Mix.......................................................................................................50
3.16 Move Arm ............................................................................................52
3.17 Move Auxiliary Station Operation ........................................................53
3.18 Move Shuttle .......................................................................................54
3.19 Move Stage .........................................................................................54
3.20 Nitrogen Dry ........................................................................................55
3.21 Pause ..................................................................................................56
3.22 Pressure Vacuum Control ...................................................................57
3.23 Program Note ......................................................................................58
3.24 Quit......................................................................................................58
3.25 Restack Plates ....................................................................................59
3.26 Serial Dilution ......................................................................................60
3.27 Shaker Nest ........................................................................................63
3.28 Shuck Tips ..........................................................................................63
3.29 Shuttle Layout Change ........................................................................64
3.30 SPE Vacuum Box Operation ...............................................................65
3.31 SPE Pressure Vacuum Operation .......................................................66
3.32 Stacker Operations..............................................................................67
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TABLE OF CONTENTS
3.33 Target Method .....................................................................................68
3.34 Timed Dispense ..................................................................................70
3.35 Vacuum Box Operation .......................................................................71
3.36 Vacuum Control...................................................................................72
3.37 Wash ...................................................................................................73
3.38 Program Operations Common Options ...............................................75
3.38.1 Additional Options .................................................................75
3.38.2 Dip Tips .................................................................................76
3.38.3 Select Shuttle Position...........................................................77
3.38.4 Stage Operations...................................................................77
3.38. Touch Off.................................................................................79
4.0 Database Editor ...............................................................................................80
4.1 The Menu Bar .......................................................................................82
4.2 The Plate Panel.....................................................................................83
4.3 The Fixture Panel ..................................................................................84
4.4 The Touch Off Panel .............................................................................85
4.5 Importing Plates-Reservoirs-Fixtures ....................................................85
5.0 Options ............................................................................................................86
5.1 User Preferences ..................................................................................86
5.2 Default Plates........................................................................................88
5.3 Default Reservoirs.................................................................................90
5.4 Configure Model....................................................................................91
5.5 Configure Com Port ..............................................................................92
5.6 Set Arm Position Names & Heights.......................................................92
5.7 View User Preferences..........................................................................93
5.8 Displayed Fixtures.................................................................................94
5.8.1 Edit Display and Order of Fixtures .........................................95
5.9 Current File Setting Option....................................................................96
5.9.1 Logging Options.....................................................................96
5.9.2 Runtime Database Usage Options ........................................97
6.0 Utility ................................................................................................................98
6.1 Shuttle/Stacker......................................................................................99
6.2 Stage...................................................................................................100
6.3 Pinch Tubes ........................................................................................102
6.4 Pump...................................................................................................104
6.5 System ................................................................................................105
6.6 Trap Operations ..................................................................................105
6.7 Miscellaneous .....................................................................................107
6.8 Arm – Vacuum Box – Aux. Station ......................................................108
6.9 Status ..................................................................................................109
6.10 Pump Calibration Tables ...................................................................109
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TABLE OF CONTENTS
7.0 Tools ..............................................................................................................113
7.1 Tools Serial Dilution Wizard ................................................................113
7.1.1 Setup ...................................................................................114
7.1.2 Set Source Row/Column......................................................115
7.1.3 Set Sample Aspirate Volume & Height, Row, and Column
Offsets ............................................................................................116
7.1.4 Set Initial Dilution Row/Column............................................117
7.1.5 Serial Dilution.......................................................................117
8.0 Technical Tips................................................................................................118
8.1 Introduction .........................................................................................118
8.2 Pre-Planning .......................................................................................118
8.3 Labware ..............................................................................................118
8.4 Labware Location................................................................................118
8.5 Writing The Protocol............................................................................119
8.6 System Parameters.............................................................................121
9.0 System Parameters – Password....................................................................121
9.1 System Parameter – Stage .................................................................122
9.2 System Parameters – X Shuttle ..........................................................122
9.3 System Parameters – Board A Misc....................................................123
9.4 System Parameters – Pump ...............................................................123
9.5 System Parameters – Y-Shuttle ..........................................................124
9.6 System Parameters – System Info ......................................................124
9.7 System Parameters – Shuttle Alignment Utility ...................................125
9.8 System Parameters – View System P.................................................126
9.9 Vacuum Calibration .............................................................................127
Section IV
Pipettor Heads....................................................................................................128
Quadra 4 Air Displacement Pipettor Heads .........................................................128
1.0 Quadra 450µL Disposable Tips ..........................................................128
1.1 Organic to Aqueous Pipetting.............................................................128
1.2 Non Contact Dispensing.....................................................................129
1.3 Clearing Tip Residual .........................................................................129
2.0 Quadra 450µL Tip Design .............................................................................129
2.1 Loading the 450µL Disposable Tips ...................................................129
2.2 Adjusting Tip Load Switch ..................................................................130
3.0 Quadra Positive Displacement Pipettor Heads .............................................131
4.0 Quadra 4 DMV Head (Disposable Multiple Volume127 ................................132
5.0 Setting the Zero Volume Adjustment for Air Displacement Pipettor Heads...135
6.0 Quadra Shuttle..............................................................................................137
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TABLE OF CONTENTS
7.0 Stackers ........................................................................................................137
8.0 Quadra Stage................................................................................................139
Section V
Suggested Pipetting Methods & Recommendations Aspirating ...................140
1.0 Pipeline Pipetting......................................................................................141
2.0 Dispensing................................................................................................141
3.0 Mixing.......................................................................................................142
4.0 Titration ....................................................................................................142
5.0 Bubbling ...................................................................................................142
6.0 Injection ....................................................................................................142
Section VI
Pipetting with the Positive Displacement Pipettor Heads ..............................142
1.0 Pipetting 0.5µL With Positive Displacement .......................................143
2.0 Dispensing 0.5µL Spots – Aqueous....................................................143
3.0 Dispensing 0.5µL Spots – DMSO .......................................................144
4.0 Protocol Verification............................................................................144
Section VII
Bioanalytical Applications ................................................................................145
1.0 Aqueous and Organic Pipetting...........................................................145
2.0 Protein Precipitation, Liquid/Liquid, and SPE ......................................145
3.0 Protocol or Program Security ..............................................................146
4.0 Calibration for the Quadra 4 ................................................................146
Section VIII
Quadra Accessories ..........................................................................................150
1.0 Custom Reservoirs.................................................................................150
Keeping Participate Matter in Suspension
Stirring Reservoir
Constant Level Reservoir
Shaker Nest
Live Bottom Reservoir
Thermal Retention Reservoir
Peltier Nest for Heating and Cooling
Vacuum Box
Filtration System for Quadra 4
Air Regulator Vacuum Trap
Fully Automated Filtration System
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TABLE OF CONTENTS
Section IX
Pipettor Tips .......................................................................................................154
1.0 Tip #1 ..................................................................................................154
2.0 Tip #2 ..................................................................................................154
3.0 Tip #3 ..................................................................................................154
4.0 Tip #4 ..................................................................................................154
5.0 Tip #5 ..................................................................................................155
5.1 Auxiliary Connector Panel ........................................................155
5.2 Serial In ....................................................................................156
5.3 Barcode Logging Functions ......................................................156
Section X
Calibration of the Quadra 4 450µL 96 Position Pipetting Head......................157
1.0 Equipment Required............................................................................158
2.0 Weighing Vessels................................................................................159
3.0 Evaporation .........................................................................................160
4.0 Pipetting To Weighing Cycle Time ......................................................161
5.0 Setup Procedure .................................................................................161
6.0 Adding the New “Calibration Curve” into the Quadra Command System
Software from Data Points...................................................................162
7.0 “Calibration Template – Q4” Spreadsheet ...........................................163
8.0 Creating a New “Calibration Curve” with Water...................................164
9.0 Creating a “Verification Curve” ............................................................166
10.0 “Calibration Template – Q4” For the Verification Curve ………..…..169
11.0 Calibration Instructions for the Quadra 4 Series 450µL Pipetting
Head………………………………………………………....................169
11.1 Definitions ........................................................................................169
11.2 Liquid Used for Calibration Set Volumes...........................................169
12.0
Liquid Used for Calibration Set Volumes…………………………….169
12.1 Importing an Existing “Calibration Curve”……………………170
Section XII
Some Properties of Liquids that Affect Accuracy...........................................171
1.0 The Relationship Between Liquid and Gaseous States of a Solvent
Have an Effect on Pipetting .................................................................171
2.0 Some Other Properties of Liquids that have an effect On Pipetting ....171
3.0 To Compensate for these Properties of Liquids, the Aspirate and
Dispense Steps have Options to Improve Performance......................172
Section XIII
Head Removal, Installation and Alignment......................................................173
1.0 Top Back View ....................................................................................174
2.0 Top Back View Open Panel.................................................................174
3.0 Head Installation..................................................................................176
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TABLE OF CONTENTS
4.0 450µL Back Fill Head Front View ........................................................177
5.0 450µL Back Fill Head Back View.........................................................178
6.0 Tip Block Assembly .............................................................................179
7.0 350µL Head C0SOS691......................................................................180
8.0 384sv Tip and Seal Assembly .............................................................181
Section XIX
Routine & Preventive Maintenance By Operator.............................................182
1.0 Lubrication Kit......................................................................................182
2.0 Cleaning the Pipettor Tips ...................................................................183
Fuse Replacement...............................................................................................184
Warranty ..............................................................................................................185
Return Policy........................................................................................................185
Quadra 4 Decontamination Form.........................................................................186
Tomtec Contact Information – If you need Help...................................................187
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PREFACE
This manual provides general information, installation and operation instructions for
the TOMTEC QUADRA 4®.
Every effort has been made to avoid errors in text and diagrams, however, TOMTEC
assumes no responsibility for any errors or omissions which may appear in this
manual.
It is the policy of TOMTEC to improve products as new techniques and components
become available. TOMTEC reserves the right to change specifications at any time.
We welcome your comments on this publication.
TOMTEC, INC.
1000 Sherman Avenue, Hamden, CT 06514 USA Phone: (203) 281-6790
Fax: (203) 248-5724 – 1-877-866-8323 – www.tomtec.com
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Tomtec
UNPACKING INSTRUCTIONS
Carefully remove the Quadra 4 from the shipping carton and place on a level surface.
When moving the equipment, lift only in the locations indicated by Figure A. Do not lift or
handle the equipment by the X-Y table.
LOCATING THE QUADRA 4
When selecting a suitable place to set up your Quadra 4, please consider the
following:
The Quadra 4 should be placed on a level surface. A slanted surface can cause the
liquid to pool to one side of a well or reservoir and may impair your ability to fully
pipette out small volumes.
Be sure to leave at least four inches (10 centimeters) between the back of the
Quadra 4 and any wall or shelving that may impair the movement of air through the
cooling fans.
Although the Quadra 4 can supply its own air pressure, some accessories may
require a water source (normally DI), a vacuum source, a place to drain waste liquid,
or additional space for a vacuum trap or controller module.
For overall space requirements, please refer to the instrument dimensions under
"Technical Specifications" on page 1:1.
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Section I General Information
Caution
Specifications & Technical Information
Warnings, Cautions, Notes, and Symbols
Quadra 4 - Pictorials
Quadra Command System 4 Software
Quadra 4, Front View
Quadra 4, Front Left View
Quadra 4, Front Right View
Quadra 4, Back Left View
Quadra 4, Back Right View
Operational Functionality
Basic Information
Connector Panel
Front Panel Buttons
CAUTION: IF THE EQUIPMENT IS USED IN A MANNER NOT SPECIFIED
BY THE MANUFACTURER, THE PROTECTION PROVIDED BY THE
EQUIPMENT MAY BE IMPAIRED.
The Quadra 4 is designed for use with various liquids, aqueous
or organic solutions with a pH near neutral. As the pH of the
solution moves away from neutral, various parts may be
affected, depending on the length of time they are in contact.
The user has full responsibility for the selection of compatible
liquids and for thoroughly rinsing the system after use. Damage
resulting from improper use of liquids and lack of cleaning are
the user's responsibility and may void the warranty.
Important Note:
Powered accessories such as Vacuum Traps, Autotraps and
Shaker Nests, should not be connected with the Quadra 4
powered on. Please turn off AC power BEFORE connecting
these accessories.
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SPECIFICATIONS
QUADRA 4
Environmental Specifications
Operating environment
INDOOR USE ONLY
Temperature
Humidity:
5ºC to 40ºC (41 ºF to 104 ºF
20%-90% (no condensation)
Storage environment
INDOOR USE ONLY
Temperature:
0 °C to 60 °C (32 °F to 140 °F)
Humidity:
5%-90% (no condensation)
Physical Specifications
Dimensions:
Width:
Depth:
Height:
Weight:
Shuttle Extension:
Shuttle Extension:
30” (762 mm)
21” (534mm)
33” (838mm)
200lbs (90.71g)
9.5” (241mm) L/R
6” (153mm) (Front to Back)
Electrical Specifications
Power requirements:
100VAC NOM, 60Hz @ 4.0 AMP
105-125VAC 60Hz @ 4.0 AMP
230-240, 50Hz @ 2.0 AMP
Pneumatic Specifications
Internal Compressed Air
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WARNINGS, CAUTIONS, NOTES & SYMBOLS
Throughout this manual, there may be blocks of text printed in bold type within boxes
or italic type. These blocks are warnings, cautions, and notes, and they are used as
follows:
WARNING: A WARNING INDICATES THE POTENTIAL FOR BODILY HARM
AND TELLS YOU HOW TO AVOID THE PROBLEM.
CAUTION: A CAUTION INDICATES EITHER POTENTIAL DAMAGE TO
HARDWARE OR LOSS OF DATA AND TELLS YOU HOW TO AVOID THE
PROBLEM.
NOTE: A NOTE indicates important information that helps you make better
use of your Quadra.
In this manual, there may be symbols within triangles, and they are used
as follows: The lighting flash with arrowhead symbol, within an equilateral
triangle, is intended to alert the user to the presence of un-insulated
"dangerous voltage" within the product's enclosure that may be of
sufficient magnitude to constitute a risk of electric shock.
The exclamation point within an equilateral triangle is intended to alert the
user to the presence of important operating and maintenance (servicing)
instructions in the literature accompanying the instrument.
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Tomtec
Introduction
The TOMTEC Quadra 4 is a third generation liquid handling workstation with
numerous optional pipetting head configurations available with or without stackers,
formatting stage selections and protocol specific accessories. Your local TOMTEC
Sales Engineer or our Worldwide Distribution Network can offer customer assistance
in selecting the correct model, pipetting head and accessories to fit individual
requirements.
The Quadra 4 incorporates an embedded PC, 15" full Color Display, our unique
Quadra 4 Command Software, and wireless keyboard. In addition, the embedded
PC provides 4 USB ports and internet network capabilities.
This manual will describe the standard Quadra 4 Pipettor - both with or without
stackers. Additional pipetting head configurations and stage options are outlines in
APPENDIX A at the end of this manual.
An electronic copy of this manual is provided in MS Word and is resident on the
internal hard drive in the instrument.
QUADRA 4
Login Instructions
The Quadra 4 uses the login functions available with the MS Windows XP Operating
System installed in the embedded PC. This feature gives the user the option of
setting operational privileges and parameters for each individual using the Quadra 4.
The Quadra 4 shipped to you has two user selectable login defaults:
administrator (PW = password)
tomtec (PW = Tomtec)
Privileges of defaults:
administrator – full application and MS Windows privileges
tomtec
-operation of application programs
-create application programs
-upload and download USB ports
The Administrator can add, delete, change users, functionality, passwords, etc. by
using the MS Windows XP functions (control panel – user accounts).
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QUADRA 4 (with stackers)
Aux. Pressure
1,2,3
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QUADRA 4 (without stackers)
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QUADRA 4
(Front Access Panel)
Recommended to be used by TOMTEC Service Technicians only!
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COMPRESSOR & PNEUMATICS
Pneumatic Panel Description
The front panel provides manual access to the pneumatic system. The pressure gauge
may be connected to several ports for readout.
Compressor Pressure:
30psi for 60 Hz operation.
25psi for 50Hz operation.
NOTE: Due to normal compressor pulsing, do not leave the pressure gauge connected
to the compressor for long time periods. This affects the life of the pressure gauge.
Blowout Pressure:
This is set by the regulator on the right hand side of the
Quadra 4. It will range from 0psi to 5psi. A more accurate
pressure reading is available on the front display.
Elevator
Pressure:
At system pressure (not a user adjustment).
Regulatory
Accessory
(API)
Pressure:
This pressure is set by the R2 pressure regulator.
It must be less than 10psi or it will rupture the bladder in the
various nests.
Switches:
Elevator Override - Actuates the stacker elevators.
Compress
orBypass:
Shuts off the internal air compressor and connects
system to external air supply. External air supply
must not exceed 35psi.
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Section II
OPERATIONAL FUNCTIONALITY
PIPETTING SYSTEM
The pipettor pistons are driven by a software controlled stepper motor. To provide
precise control it requires 400 half steps of the motor to make one revolution. This in
turn drives the syringe pistons aspirating or dispensing fluid. On the 96SV and 384 head
the pistons extend to the bottom of the pipette tips. This provides positive displacement
pipetting, essential for sub-microliter dispensing. The 450µL disposable tip head is an air
displacement system, providing up to 450µL dispensing.
Upon initialization, the stepper motor drives the piston up to actuate a photo sensor.
This is the home position for the volume counter. It then counts down to the zero volume
position for the pistons. Aspirate and dispense volumes are made by counting up or
down. You must always dispense an equal volume that has been aspirated. If you try to
dispense more than you have aspirated, "Pump Range" error will occur. If you do not
dispense the full amount aspirated, the accumulation will again cause "Pump Range"
error.
There are four speeds for the head motor. Always use speed 3, unless you are aspirating
or dispensing viscous liquids. These may not flow through the tip orifice in the time allowed
at speed 3, especially, with the 0.015 inch diameter of the small volume tips on the 96SV
and 384 head.
The 450µL head has the ability to count below zero volume to remove the tips. There is a
software number that determines this function. It drives the tip stripper plate down to or
below the "O" ring to strip the tips from the tip pins. Each time the stripper plate moves
down, it applies a very light coating of lubricant to the tip pin "O" rings.
When loading tips, a sensor detects whether the tips have been fully sealed, thereby
preventing error in the pipetting volumes. If this sensor does not actuate, the stage will try
three times to load the tips properly. If it fails after the third time you are asked to use the
manual lubricator on the tip pin "O" rings. If this occurs frequently it is time to replace the
internal lubrication pad.
The design of the Quadra tips is unique. The long narrow portion (90µL) allows pipeline
pipetting, permitting you to aspirate several reagents sequentially using small air gaps for
separation. This, combined with the multiple stations on the Quadra 4 shuttle, means all of
the reagents can be added in one pass. The larger volume buffer can be used to wash out
the smaller volume of the compound. The 450µL volume of the Quadra tips also permits the
larger tasks, such as Solid Phase Extraction, Liquid/Liquid Extraction, or Protein Precipitation.
The Quadra tips (PN 196-205) are available with a standard orifice size 0.036”, as both
sterile, and non sterile. The Quadra tips are retained in their rack. This permits handling
them as a single unit, not individual tips. They may be used, shucked as a unit, and
reused.
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QUADRA 4
GENERAL INFORMATION
Tomtec uses a different design philosophy on the Quadra 4 compared to most
competitive multi-well pipettors. Rather than move a complex, heavy, pipetting head
over the microplates, the Quadra design fixes the head firmly in one place. The shuttle
is used to move lightweight plates and reservoirs to the stationary head. This results
in a very reliable design. All of the electronic control wiring is fixed in place. Moving
wiring is subject to breakage. An intermittent breakage in a control line is extremely
different to diagnose. This has the additional advantage of being able to easily
change the pipetting head for very different applications. Four bolts and a plug in
connector complete the change over.
The 6-station shuttle, under software program control, can quickly position any station
under the pipettor head. More importantly, the shuttle can be automatically loaded and
unloaded via the stackers for those applications requiring the processing of more than
6 devices. Thus, in comparison to large flat bed pipettors, the Quadra design can
handle 50 devices, plates and reservoirs, to and from the pipettor with walk away
automation.
A stage is provided under the pipettor head to lift the various devices to the tips for
precise vertical control. Depending on the application, this can be a simple stage with
touch off control or it may be a full indexing stage for reformatting or serial dilution
testing protocols.
The straight forward design of the Quadra 4, lends to very easy and intuitive software
control. Complex protocols requiring various microplates and reservoirs can be
created to run in a walk away mode of operation, while only requiring 30 inches of
bench top space.
Tomtec has elected to supply the Quadra 4 in a variety of configurations. This is to
allow the end user the freedom of selecting the most cost effective model for their
specific requirements. This operator's manual describes the various elements that are
available. Not all of these features are on all models.
This is our top of the line model. It includes stackers, an indexing stage, and has a full
pneumatic system for use with additional accessories.
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QUADRA 4 PRODUCT LINE
QUADRA 4 NS
This is the Quadra 4 without the stacker option but with the
indexing stage for reformatting i.e., 96 to 384 to 1536. It does not
have the full pneumatic system of the Quadra 4.
QUADRA 4 SPE
Similar to the Quadra NS, this model is also without stackers.
The indexing stage has been replaced with a touch off stage.
A manually operated vacuum box and vacuum trap is
included. The Quadra 4 SPE is designed for solid phase
extraction applications. It has the ability to blow nitrogen
through the tips for sample dry down. It has a limited
pneumatic system.
QUADRA 4 LTD
This is the same as the Quadra 4 SPE Model except it
does not have the vacuum box and vacuum trap
included.
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Quadra Command
System v4
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Section III
Quadra 4 Command System Operators Manual
OM400-730 REV G
Current Version Level 4.6.x
Copyright (c) 2008. All rights reserved.
TOMTEC, Inc., 1000 Sherman Ave., Hamden, CT 06514
Phone: (203) 281-6790 Fax: (203) 248-5724
http://www.tomtec.com
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Tomtec
Quadra
Quadra Command
System v4
1.0 Introduction
1.1 What's New
Some of the new features in the Quadra 4 Command Software v4.x are:
Version 4.7.7.30
• The number of Tip Loads is now displayed on the main screen. (See description
below)
Version 4.7.7.29
• The ability to validate the processing of mirror racks which have been processed
with the Tomtec PSID2 software. (See description below)
Version 4.7.7.xx
• Programs written for the Quadra 3 will run on the Quadra 4 without modification.
• The Quadra 4 SPE is now a selectable model.
• Pinchtubes can be cycled in both the tip load and tip shuck functions.
• Support for the "DMV" head added(Version 2.x.x firmware).
• "Target Method" function added.
• Shuttle alignment "wizard" replaced with an utility(Version 2.x.x firmware).
• Detailed logging of "Large" macro functions.
• Tip Load and Shuck heights can now be set in the database(version 2.x.x
firmware).
• Pump calibration data entry function now a part of the GUI.
• Pump calibration tables can now be viewed, printed, and saved to a file.
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1.1.1 Tip Load Counter
A counter has been added to the main screen which displays the total number of Tip
Loads. This can be used to determine if it is time to lubricate the “O” rings.
The counter is displayed in the status bar at the bottom of the main screen as shown:
The counter can be reset by using the System → Reset Tip Load Count function. This
should be reset when the “O” rungs are replaced or lubricated.
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1.1.2 PSID2 Mirror Rack Checking
If Black and White Mirror Racks are being used in a protocol which have already been
processed with the Tomtec PSID2 device, the Quadra 4 can be setup to validate the
handling of these racks to ensure that they are being processed correctly.
The Quadra 4 software can validate that:
1.
The Mirror Racks and the Destination Plate have been placed onto the Quadra in
the correct orientation.
2.
The Black Mirror Rack, the White Mirror Rack, and the Destination Plate
barcodes all match the values specified within the PSID2 software.
In order to use this validation, a barcode reader must be installed on the Quadra 4 on
the RIGHT side of the pump.
Transferring the PSID2 Files
To begin, the Completed Main Operating File(s) need to be copied to the Quadra 4.
The files are saved on the computer running the PSID 2 Software in the directory:
C:\tomtec.psid2\psid2\archive\
Only the COMPLETED CSV files should be copied. These files have the naming
convention of:
<DestinationPlateBarcode>_MOF_<BlackRackBarcode>_<WhiteRackBarcode>_<Date/Time>.csv
These files should be copied to the directory on the Quadra 4,
C:\Program Files\Tomtec\Quadra 4 Command System v4\PSID2\
Quadra 4 Setup
Before running the protocols on the Quadra, the Mirror Rack Checking needs to be
enabled.
In the Options → User Preference dialog, set the “Mirror Rack Checking” value to
“enabled”.
In the File → Log Settings dialog, ensure that the following settings are being used:
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and use the “Set as Default” button to save these settings.
The shuttle positions of the Black and White mirror racks should be specified (see
below), so the Quadra code knows where to look for each of these racks.
Running a PSID2 protocol
Running a protocol with Mirror Rack Checking enabled is done the same way as
running any protocol on the Quadra 4. However, during the run, the Quadra software
will scan the barcode of the incoming rack and look to find the corresponding PSID2 file
in the above directory. The software will then check the barcode of the other mirror rack,
and the destination plate to ensure that they match the expected barcodes and that they
are oriented correctly on the Quadra shuttle. If no barcodes are read, or if an
unexpected barcode is encountered an error message will be displayed, and the user
will be asked to fix the problem and retry.
The Mirror Rack Checking code is looking for a single Aspirate from the White Mirror
Rack, a single Aspirate from the Black Mirror Rack (not necessarily in that order), and
one or more dispenses into the Destination plate. Once all of these requirements are
met, the csv file is moved into a sub directory named “Completed” and the Quadra will
no longer check barcodes.
In some instances, it may be necessary to have additional steps within the protocol
when processing the Mirror Racks. Any additional steps in the protocol which are not
related to the standard mirror rack checking should have the "Disable Barcode Reading"
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option checked. This will cause that step to be ignored by the Mirror Rack checking
code.
Also, it is important to note that the barcode will only be read correctly if the stage is all
the way down before the step starts. In some cases it might be necessary to set the
option to "Drop Stage after Dispense" in the step BEFORE the step which checks the
barcode.
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1.2 Quadra 4 DMV Head
Quadra 96 DMV Pipettor Head
The new Quadra 96 DMV pipettor head adds increased flexibility to
Tomtec’s Quadra 4 product line. The DMV (Disposable Multiple Volume)
pipettor head will accommodate 4 different sets of pipettor tips. A 250µL, a 50µL
tip, a 20µL tip, and a 250µL tip with a wide bore. All are available from Tomtec.
Each rack of tips are interchangeable, even within a pipetting protocol. It allows the
user to select the appropriate tip for each specific application. The use of
disposable tips requires the use of an air displacement pipettor. The liquid
aspirated in the tip must never contact the pipettor piston. As such, the minimum
dead air gap must be equivalent to the tip volume, i.e. a 250µL tip always has a
minimum of 250µL of air between the liquid being aspirated and the face of the
pipettor piston. Air displacement pipettors are defined by the General Gas
Law PV=?RT. When aspirating the same gas (air) at the same temperature,
the equation becomes P1V1=P2V2. When aspirating the maximum tip volume, the
aspirating vacuum within the tip is limited to half of atmospheric pressure.
For volumes less than the maximum tip volume, the aspirating vacuum is
proportionately less. Thus, at volumes less than 10% of the tip volume, air
displacement pipettors loose accuracy. There is insufficient negative pressure to
overcome, the resistance to flow within the tip itself. These are the forces of
capillary action. Thus, the ability to choose the right tip for the volume to be dispensed,
increases operational accuracy.
The Quadra 4 stacker design allows the appropriate tip rack to infeed to the shuttle and
then outfeed back to the stackers. The further ability of the Quadra 4 stacker design
allows restacking, which provides the means to sort incoming tip racks, microplates, and
reservoirs, as they come and go to the shuttle for processing. The flexibility of operation
is left solely to the creativity of the operator in structuring the desired protocol.
As with the incoming microplates and reservoirs, the Quadra 4 requires the database
value of the tip or the other devices that are on the shuttle at any given time. This is
accomplished with the shuttle layout icon. Each time a different plate, reservoir, or rack
of tips comes on the shuttle, the shuttle layout must agree with the devices that
are in place. A change in layout is only required if a different device is used. It is not
required if the same devices is simply exchanged, i.e. one rack of 250µL tips for another
rack of 250µL tips. The database values, for each device, provides the essential
properties of the device, i.e. top of well,
bottom of well, tip volume, etc.
The tip racks for the DMV tips are all of a common size with the SBS footprint. The tip
volume is a database value that describes the pipetting limits. The 250µL and 50µL tips
are of the same length, however, the 20µL tip is a shorter length. The database value
provides that correction. The 50µL tip has a longer narrow cannula. Depending on
the liquid being handled, it may be necessary to use a slower aspirate speed.
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This is to assure that the liquid level in the tip is following the piston motion.
The longer cannula of the 50µL tip, may
restrict the flow of more viscous liquids, such as plasma.
To increase the accuracy of air displacement pipetting at smaller volumes, the
Quadra 4 provides 2 distinct pipetting features. The first is the ability to blow
through the tip with low air pressure. This is the Quadra’s timed dispense
function. At the conclusion of the dispense motion, low positive pressure air (1 to
5 psi) may be blown through the tip orifice. This effectively clears the tip orifice of
the variable volume that may be retained by capillary action. The 1µL, that may, or may
not be retained by capillary action, may only be 1% of a 100µL dispense, but it is 10% of
a 10µL dispense. The use of the timed dispense function is far more effective at
consistently clearing the tip of residual than the customary use of piston blow
out air gaps.
The second function feature is target dispensing to improve accuracy at a low volume.
The target method may be selected on any aspirate/dispense function. There are
two phases to the target method. Experience and collected data has shown that
pre-wetting the tip, prior to aspirating the sample volume, improves pipetting accuracy at
the small volumes. When target method is selected, the tips are automatically prewetted by aspirating and dispensing 30µL back to the source. For dispense
volumes less than 50µL, the second step is to aspirate 50µL then automatically
dispense back to the source all but the target volume.
Assume 10µL is the target volume to be dispensed. 30µL is first aspirated and
dispensed back to the source to pre-wet the tip interior. Then 50µL is aspirated and
40µL is dispensed back to the source leaving that target volume of 10µL in the tip.
What is the purpose of this motion? As stated earlier, by aspirating 50µL, a higher
internal vacuum is created in the tip, than if only 10µL was aspirated. This higher
vacuum is more effective in filling the tip with the desired liquid aliquot by overcoming
the resistance to flow at the small orifice. Next, the pipette piston reverses to the
dispense function, thereby forcing the excess volume out of the tip, leaving the
requested target volume to be dispensed to the destination.
The last part of the target method pipetting occurs on the dispense at the destination.
The pipettor piston moves to dispense all but the last 3µL in the tip. The last 3µL are
then ejected with a positive pressure of 2-3 psi through the tip orifice. The imparted
kinetic energy, to the remaining 3µL slug of liquid, causes it to eject cleanly from the tip
orifice.
The 3µL value was arrived at empirically. Initial testing left 8µL to be ejected by
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air pressure. There was not enough velocity applied to the liquid slug which allowed
some of it to cling to the tip exterior on the dispense. The 3µL volume gave better
results for the reason stated.
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1.3 Getting Started
The above stated pre-wet and 50µL aspirate for the target method are based
on the 250µL tip. The same value would apply when using the 50µL tip i.e., 30µL
pre-wet, 50µL then dispensed to the target volume. When the 20µL is used, the prewet and put back volumes would be the full tip volume of 20µL.
Creating a stacker program on a Quadra 4 is akin to a software game. There
are certain rules that apply. The objective is to create a smooth running
program that can be repeated on demand. The real advantage of automation
is to create a specific protocol, that will run precisely the same, every time. This
allows small optimizing modifications that will be faithfully reproduced to enhance the
end results. Once the program is created, anyone can execute it. The stackers are
loaded in the defined order of the program and then executed precisely the same
every time the protocol is run.
The basic rules are as follows: First, an understanding of stacker access by
the shuttle stations. Shuttle station 2 is the most flexible. It can access all four
stackers. Stations 1 can access the Left Rear Stacker (LRS) and the Left
Front Stacker (LFS). Station 3 can access the Right Rear Stackers (RRS) and the
right front stacker (RFS). Station 5 can access the LFS and RFS. Stations 4 and 6 can
only access the LFS and the RFS. Thus, they are the least useful. Therefore, they are
commonly used for fixed functions, such as the Ultrasonic Tip Wash Station or a
vacuum box. They are also useful for a common reservoir that is used through out the
protocol.
While each pipetting protocol may be different, in general terms, it is effective to use
the left rear stacker for the incoming source plates, or reservoirs and the right rear
stacker for the destination plates. In this manner at the end of the sequence, the
source plates, or reservoirs may be restacked for the next set of destination plates.
The tip boxes are then interspersed, depending on whether they are to be assigned
and reused with the specific sources or not. A distinct advantage of the Quadra 4 is
the ability to make a non-contact dispense. The target method may be used to make a
clean dispense at the top of the well, without touching the contents of the receiving
plate. This allows assigning a rack of tips for use with specific reagents,
without fear of cross contamination.
As the pipetting protocol become more complex, moving microplates, reservoirs and
tips in and out of the stackers to the shuttle becomes more challenging. While
everybody finds their own best way, a starting point is to step the protocol through the
entire program, step-by-step using the simple Quadra programming features. It
provides a visual presentation of the program as you proceed.
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The program allows you to assign a descriptive name to each plate, reservoir, or rack of
tips. This aids in tracking the program. As each new device is placed on the shuttle, use
the shuttle layout icon to bring the appropriate database values. At the end, it is essential
that you record the starting location of each microplate, reservoir, or tip rack in
their respective stackers. This information may be recorded in an initial pause
statement.
Tomtec personnel are available to assist you in creating a stacker program.
When using this program for the first time:
1. The Calibration Height is set on all fixtures in use. These values can be
set in the Database Editor. Open the Quadra DBEditor by selecting the
‘Database’ Menu > ‘Edit Plate, ... Database.’
2. The COM Port setting is correct. This can be set by going to the Options Menu >
Quadra > Configure COM Ports.
3. The proper machine model is selected. This can be set by going to the Options
Menu > Quadra > Configure Model.
Upon Connecting to the Quadra for the first time, the program will automatically recognize
the head configuration, and change its settings accordingly. Make sure, before you
start writing your protocol, that you have Configure Model set to the right
configuration, or simply connect to the machine before programming.
Preparing the Quadra to run a program:
1. Click the ‘Connect’ button on the Main Toolbar to establish the serial connection
with the Quadra.
2. After the connect operation is successful, the ‘Initialize’ button will enable. Click this
button to initialize the Quadra.
3. Be sure to include the proper fixtures on the shuttle layout before adding steps to
your program.
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2.0 Main Screen
Labware - Components Box
Program Operations Box
Program Data Box
Shuttle Position Data
2.1 Labware - Components Box
Icons represent the various plates and fixtures available. Use the mouse to
configure the shuttle. Click on the appropriate icon and drag it to the desired
position. First populate the shuttle with fixtures. Plates, reservoirs, and tips are
then added to fixtures already present on the desired shuttle position.
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2.2 Program Operations Box
Icons represent the various available functions for the current model configuration and
are used to generate the program steps in the adjacent list box. Typically to generate a
program step:
1. Select the shuttle position by single clicking on the shuttle graphic with the left
mouse button.
2. Select the icon for the desired function by single clicking with the left mouse button.
3. Enter information into the dialog boxes and select OK.
2.3 Program Data Box
The vertical progress bar indicates volume in the tips. When designing a program, it will
show the current volume in the tips as if the program has run to the currently
highlighted step (volume is assumed zero at the start of the program). When
running a program it will show the actual volume in the tips. White sections represent
volumes designed to be air-gaps. Blue and light blue sections represent liquid volumes,
aspirated separately. If "ERROR" is displayed, then either an over and under volume
condition has occurred.
Other information indicated here includes the status of simulation mode as well as loop
counts.
2.4 Shuttle Position Data
This dialog box is accessible through right or middle mouse button clicks on
the shuttle position. Additionally user preferences can be modified to make this
screen appear when a fixture or plate/reservoir is placed on the shuttle on the main
screen.
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Labware indicates the current fixture on the shuttle position.
Component indicates the current plate or reservoir on the fixture.
Name indicates a user specified description of a shuttle position. Typically this is used
for describing the liquid contents of a plate or reservoir. If Name is not blank then it will
be used in place of the labware/components to describe the shuttle position on the main
screen and in the users program.
Notes allows for any addition comments regarding the shuttle position. It is only
displayed in this dialog box.
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3.0 Program Operations
3.1 Arm Plate Transfer Operation
The arm plate transfer operation is used to get plates from and put plates to a stacker
nest, vacuum box, or auxiliary station.
Conditions
Requires a ‘plate capable’ fixtures such as a stacker nest. Only shuttle positions 3 and 4
are valid for the Quadra 4.
Remarks
Set Transfer Type indicates whether the hand is getting or putting the plate.
Get Plate From / Put Plate To indicates whether the plate is being transfer from/to the
shuttle or auxiliary station.
Select Shuttle Position Button is used to change the shuttle position that should move
under the arm. The number on the button indicates the currently selected position (see
Select Shuttle Position ).
Set Arm Height indicates the position to grab / release the plate. Preset
positions are available by clicking on the appropriate button. Preset buttons can be
set under Options - Accessories - Define Arm Positions (see Set Arm Position Names
and Heights).
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3.2 Aspirate
To Aspirate is to pipette or draw from a designated plate or reservoir into the tips.
Conditions
To Aspirate, you will need to select a shuttle position that contains a plate or
reservoir capable fixture.
Remarks
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Air Gap Volume
Air Gap Volume is the amount of air (in µL) that is aspirated in the tips before any
liquid
is aspirated. This allows you to blowout any residue at the end of your dispense in
order to have the least amount of error when calculating the amount of liquid actually
dispensed into your plate.
The Air Gap is taken at the height set by the Clear Top Steps in your plate
database. The height of the blowout is set in the Dispense Dialog when you
insert a Dispense Step, and is NOT a result of the Clear Top Steps value.
Note: Air Gap Volume becomes the Blow Out Volume when Dispense occurs.
Aspirate Volume
Aspirate Volume is simply the amount of liquid (in µL) that is drawn into the tips. This
is the amount of liquid that you will want to Dispense in your plate(s).
Calibration Table
The Calibration Table drop down menu consists of user defined pump data for the
liquid type used. See documentation on distribution CD.
Aspirate Speed
The Aspirate Speed is the rate at which the tips will pipette the liquid. This can be a
value between 1 and 4, 4 being the fastest. By default, 3 is automatically selected.
3.3 AutoTrap Operations
This operation provides control for the AutoTrap.
Conditions
None
Remarks
Turn On Filtering and Turn Off Filtering starts and stops plate filtering through a
vacuum box attached to the AutoTrap.
Check Tap and Prompt will perform a check of the liquid level in the trap. If it is full,
the program will pause and the user will be prompted to empty.
Important Note:
Powered accessories such as Vacuum Traps, Autotraps, and
Shaker Nests, should not be connected with the Quadra 4
powered on. Please turn off AC power BEFORE connecting
these accessories.
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3.4 Cell Wash
The Cell Wash function allows for a multi point aspirate of a 96 well plate.
Conditions
This function is available for 96 well plates and pumps only.
Remarks
Air Gap Volume is the amount of air (in µL) that is aspirated in the tips before any
liquid is aspirated. This allows you to blowout any residue at the end of your dispense
in order to have the least amount of error when calculating the amount of liquid
actually dispensed into your plate.
The Air Gap is taken at the height set by the Clear Top Steps in your plate
database. The height of the blowout is set in the Dispense Dialog when you
insert a Dispense Step, and is NOT a result of the Clear Top Steps value.
Note: Air Gap Volume becomes the Blow Out Volume when Dispense occurs.
Aspirate Volume
Aspirate Volume is simply the amount of liquid (in µL) that is drawn into the tips. This
is the amount of liquid that you will want to Dispense in your plate(s). Note that the
aspirate will occur at the center of the well. If you do not want to aspirate a volume, set
it to zero(0).
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Aspirate Speed
The Aspirate Speed is the rate at which the tips will pipette the liquid. This can be a
X-Y Offset Aspirate Volumes
The four aspirate volumes are offset from the center of the well in the X/Y/-X/-Y
directions. The default offset is derived from the plate database side touch number
minus 20 steps. If you want to skip any offset, enter a zero(0) volume. A zero
(0) offset will keep the tip in the center of the well.
The "Total Volume" is a summation of the air gap, center aspirate, and offset
aspirates.
Aspirate Height
Aspirate Height is the height at which the tips will aspirate relative to the stage home
position. This allows you to aspirate at the appropriate height without aspirating air.
You have the option of entering a value manually, or using the depth control to adjust
how deep you would like the tips to go into the well. For this function to work
properly, you must have the ‘Top’ and ‘Bottom’ Step values entered correctly
into the plate/reservoir database.
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3.5 Delay
To Delay is to pause the program for a specified period of time, designated in
minutes and seconds, without the need for user intervention.
Conditions
There are no requirements for this step to occur.
Remarks
Minutes
Designate the number of minutes the program is suppose to wait, in conjunction
with the number of seconds specified, before program continues.
Designate the number of seconds the program is suppose to wait, in conjunction
with the number of minutes specified, before the program continues.
3.6 De-Lidder Operation
The de-lidder operation is used to take lids off and put lids on plates.
Conditions
Requires a ‘plate capable’ fixtures such as a stacker nest. Only shuttle positions 1 to
3 are valid for the Quadra 3.
Remarks
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Take Lid OFF Plate / Put lid ON Plate indicates how the lid should be handle.
Select Shuttle Position Button is used to change the shuttle position that should move
under the de-lidder. The number on the button indicates the currently selected position
(Select Shuttle Position (see page 46).
3.7 Dispense
To Dispense is to pipette a liquid from the tips to a designated plate or reservoir.
Conditions
To Dispense, you will need to select a shuttle position. In order for the status
of the tip head to operate correctly, an Aspirate Step will have to be present before
a Dispense Step.
Remarks
Dispense Volume is the amount of liquid (in µL) that is to be pipetted into the target
plate or reservoir.
Calibration Table
The Calibration Table drop down menu consists of user defined pump data for the
liquid type used. See documentation on distribution CD.
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Dispense Speed
The Dispense Speed is the rate at which the tips will pipette liquid. This can be a
Dispense Height
Dispense Height is the height at which the tips will dispense from within the well.
This allows you to dispense at the appropriate height to wells without creating air
pockets within the wells.
A new feature that is now included in version 3.0 is the ability to view a image
representing depth control. This is viewable at the right of the ‘Aspirate Height’
value. You have the option of entering a value manually, or using the
depth control to adjust how deep you would like the tips to go into the well.
For this function to work properly, you must have the ‘Top’ and ‘Bottom’
Step values entered correctly into the plate/reservoir database.
Blow Out Volume
Blow Out Volume is simply the amount of air (in µL) at the end of your Dispense
Step in order to flush the tips of all remaining liquid.
Blow Out Height
Blow Out Height is the height at which the tips will blow out the remaining
substance from within the well.
Touch Off
While Dispensing liquid into the plates, due to the surface tension at the tip orifice,
drops may form and remain on the tips when dispensing has concluded. To
accurately pipette the correct amount of liquid into the wells, these drops must be
captured. Using the ‘Touch Off’ is one method of capturing hanging drops
remaining on the tips. By setting the correct values within the Plate
Database, you can have the tips contact the side of the wells in order to release
the drops into the wells.
The Touch Off will occur at the height at which the Dispense or Blow Out has
ended. For example, if Liquid Following has been enabled, the Touch Off will
occur at the height at which the stage has come to a rest, otherwise it will
occur at the height specified by the ‘Dispense Height’ value.
Dip Tips
Dip Tips is another method of capturing hanging drops. Dip Tips is a
vertical Touch Off. What this function accomplishes is capturing drops held on
the tips by placing the tips in very close proximity to the liquid that has already
been dispensed into the wells. The ‘Dispense Tip Dip’ Value, by default,
cannot exceed (be less than) the Dispense Height. This is to prevent the
Dip Tip function from never reaching the liquid.
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If Liquid Following is enabled, however, the restraint on the Dispense Tip Dip value is
lifted, due to the fact that the ending height after the Dispense Function has been
completed may be higher than the Dispense Height Value, or the height at
which the Dispense Function actually began. It is up to the user from this point to
estimate approximately how far within the well the liquid will begin after the Dispense
function has completed.
3.8 Empty Sample
To dispense all liquid from tips completely.
Conditions
To Empty Sample, you will need to select a shuttle position that contains a plate or
reservoir capable fixture.
Remarks
Height
Empty Sample height is the height at which the tips will dispense liquid into the well.
Touch Off
While Dispensing liquid into the plates, due to the surface tension at the tip orifice,
drops may form and remain on the tips when dispensing has concluded. To accurately
pipette the correct amount of liquid into the wells, these drops must be captured.
Using the ‘Touch Off’ is one method of capturing hanging drops remaining on
the tips. By setting the correct values within the Plate Database,
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you can have the tips contact the side of the wells in order to release the drops into
the wells.
The Touch Off will occur at the height at which the Dispense or Blow Out has ended.
For example, if Liquid Following has been enabled, the Touch Off will occur at the
height at which the stage has come to a rest, otherwise it will occur at the
height specified by the ‘Dispense Height’ value.
Dip Tips
Dip Tips is another method of capturing hanging drops. Dip Tips is a vertical
Touch Off. What this function accomplishes is capturing drops held on the tips by
placing the tips in very close proximity to the liquid that has already been dispensed
into the wells.
The ‘Dispense Tip Dip’ Value, by default, cannot exceed (be less than) the
Dispense Height. This is to prevent the Dip Tip function from never reaching the
liquid. If Liquid Following is enabled, however, the restraint on the Dispense Tip Dip
value is lifted, due to the fact that the ending height after the Dispense Function has
been completed may be higher than the Dispense Height Value, or the height
at which the Dispense Function actually began. It is up to the user from this point to
estimate approximately how far within the well the liquid will begin after the Dispense
function has completed.
Empty Speed
This set the pump speed for the empty sample function. Note that if the firmware
version is below the expected version, the pump shuck speed will be used.
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3.9 GOSUB Program
Calls a different program at that point, which will run from start to finish, and then
return to the original program, and finish the remaining steps.
Conditions
There are no physical requirements to run this step.
Remarks
File Button
The File Button calls an Open File Dialog box, which allows the user to select
another program, and will automatically insert the full path name into the appropriate
edit box.
Pause Before Executing GOTO Program
By checking Pause, the program will stop and wait for user intervention when the
program reaches this step, but before it runs. A Text box will appear after the Pause
function is selected, giving the user an opportunity to generate a message to be
displayed when the program pauses.
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3.10 GOTO Program
Calls a different program, which it will run from start to finish, and then stop.
Conditions
There are no physical requirements for this step.
Remarks
File Button
The File Button calls the Open File Dialog box, which allows the user to select another
program, and will automatically insert the full path name into the appropriate edit box.
Pause Before Executing GOTO Program
By checking Pause, the program will stop and wait for user intervention when the program
reaches this step, but before it runs. A Text box will appear after the Pause function is
selected, giving the user an opportunity to generate a message to be displayed when the
program pauses.
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3.11 Load Tips
To Load Tips is to attach disposable tips to the tip pins, or attach a fixed tip
assembly to the tip head assembly.
Conditions
To Load Tips, select a shuttle position that has a tip capable fixture.
Remarks
Load Tips
Sets the function to load tips from the selected shuttle position.
Pre-Wet Tips
Sets indicator to pre-wet tips on next Aspirate step. Pre-wet will not be
performed until start of Aspirate function. Indicator will persist between user
program runs. This options is only available for certain pumps.
Note: On models equipped with the DMV head, the tip volume is set by the selected tip
fixture. That is if a 50ul tip fixture is selected, all liquid handling functions after the tip
load will set their maximum volume to 50ul.
The firmware will set its maximum volume to 50ul. This is done to prevent any pump
range errors.
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3.12 Lockout Shuttle Position
Lockout Shuttle Position will prevent any pipetting function from occurring at that
position for a specified period of time.
Conditions
There are no physical requirements for this function.
Remarks
Shuttle Position
Select the shuttle position number to lock.
Seconds
Designates the number of seconds to prevent operation on that position.
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3.13 Loop
Enables the user to repeat a series of steps and allows the user the ability to perform
repetitive stacker functions.
Conditions
There are no requirements for this function.
Remarks
Selects stacker or stackers to infeed plates to the shuttle.
Selects stacker or stackers to outfeed plates from the shuttle.
Number Of Cycles
The number of cycles to run the enclosed steps.
Disable Lid Capability indicates that if the stacker supports lids than the plate
being processed does not have one. This is necessary for thin plates.
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3.14 Magnetic Nest
A stacker accessible nest that can be fitted with a variety of magnet
configurations for 96 or 384 well plate processing of magnetic and paramagnetic
beads/particles. The magnetic surface or probes are moved by programming the
Quadra to deliver pressurized air to the nest bladder which raises the magnetic platform
Conditions
A Magnetic Nest has to be present on the selected shuttle position.
Remarks
Select Device
The Magnetic Nest can be connected to the Quadra in one of two places: Magnetic
Nest 1 is connected to the air terminal on the left side of the machine, while Magnetic
Nest 2 is connected to the right side of the machine.
Action
Turns the units On and Off.
Toggle Action
When selected, the value will appear in red to give quick visual identification of what this
function is to perform.
Lockout Position
Locks the position from use for the given amount of time, specified in seconds.
For more information, see "Lockout Shuttle Position."
Seconds
Number of seconds to lockout the shuttle position.
Shuttle Position
Click the appropriate radio button to select the shuttle position holding the Magnetic
Nest.
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3.15 Mix
Alternating aspirate and dispense steps to achieve a uniform suspension or
thorough mixing of chemicals, reagents, and/or buffers.
Conditions
To Mix, you will need to select a shuttle position that contains a plate or reservoir
capable fixture.
Remarks
Mix Cycles
Mix Cycles is the number of repetitions the mixing process will go through before
commencing this step.
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Mix Volume
The Mix Volume is the amount of liquid (in µL) that is to be aspirated and
dispensed to the designated well for x number of cycles.
Air Gap/Blow Out
See Aspirate / Dispense Function.
Blow Out Height
See Dispense Function.
Touch Off
Dip Tips
Aspirate Speed
See Aspirate Function.
Aspirate Height
Dispense Speed
Dispense Height
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3.16 Move Arm
The move arm operation is used to change the position of the device as well
as open / close the hand.
Conditions
None
Remarks
Set Hand (before move)
Set Hand (before move) indicates the state of the hand before and during the move.
Set Arm Height
Set Arm Height indicates the absolute position for the arm to move to.
Import Auto-Arm Height
Fetches the user pre-defined arm heights.
Set Hand (after move)
Set Hand (after move) indicates the state of the hand after it moves.
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3.17 Move Auxiliary Station Operation
The move auxiliary station operation is used to change the position of the device.
Conditions
None
Remarks
Down Arrow (1) / Up Arrow (2) is used to change the auxiliary station
position under the arm. Position 1 is the rear position and position 2 is the
front position.
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3.18 Move Shuttle
Allows a simple move of the shuttle to any position.
Conditions
Remarks
Shuttle Position
Select the shuttle position that will be moved. The position selected is positioned
above the stage.
3.19 Move Stage
Moves the stage to the specified height and position.
Conditions
There are no requirements for this operation.
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3.20 Nitrogen Dry
The Nitrogen Dry operation provides a means to blow nitrogen gas out the tips
to expedite sample drying.
Conditions
A 450µL backfill pump with the optional nitrogen gas valve installed is required.
Remarks
Seconds indicates the amount of time for nitrogen gas to be blown out the tips.
Height indicates the desired stage position to perform the operation.
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3.21 Pause
Stops the program and will not continue until the user intervenes.
Conditions
Remarks
Shuttle Position
By selecting a shuttle position, the program pauses at the specified shuttle
position and waits for user intervention.
Message
In the Text Box below, the user has an opportunity to generate a message
that will appear on the screen when the system pauses.
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3.22 Pressure Vacuum Control
This function is used to preset both pressure and vacuum.
Conditions
The vacuum controlled mini-trap and software controlled pressure regulator must be
installed.
Remarks
Set Pressure
Sets the pressure regulator from 0(off) to 15 PSI.
Set Vacuum
Sets the min-trap vacuum from 0(off) to 25 inches hg. If checked, the program
will pause
for the vacuum to build.
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3.23 Program Note
This is a non-function. The "note" is a comment line added to the program for any
reason.
Conditions
None
Remarks
3.24 Quit
Stops the program and parks the shuttle at a specified position.
Conditions
Remarks
Shuttle Position
Select the shuttle position you wish to stop the shuttle at the completion of the
program.
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3.25 Restack Plates
Restack Plates is the repeated Infeed of a plate from one stacker and Outfeed
that plate to another stacker.
Conditions
A plate capable fixture must be located on a shuttle position that has access to
both the source and destination stackers.
Remarks
Selects stacker or stackers to infeed plates to the shuttle.
Selects stacker or stackers to outfeed plates from the shuttle.
Number Of Plates
The number of plates to restack.
Disable Lid Capability indicates that if the stacker supports lids than the
plate being processed does not have one. This is necessary for thin plates.
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3.26 Serial Dilution
A method of creating a numerical progression of concentrations along a series of
rows or columns by sequentially aspirating the same volume and transferring that
volume to the adjacent row or column a number of times.
Conditions
The Serial Dilution function requires a selected shuttle position.
Note: This function is only available when a removable tip pump is installed.
Remarks
Number of Dilutions
Depending on plate type and tip arrangement, the number of dilutions can range
from 1 to 24.
Mix/Xfer Aspirate Height
Aspirate Height is the height at which the tips will aspirate from the well. This
allows you to aspirate from wells filled to a variable amount, without sucking in any
air.
Mix/Xfer Dispense Height
Dispense Height is the height at which the tips will dispense into the well. This
allows you to dispense to wells filled to a variable amount, without creating air
pockets within the wells.
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Initial Dispense Volume
The initial concentration pipetted from a sample plate.
Plate Direction
The direction in which the dilution is performed. This depends on the plate and
tip arrangement.
Mix Cycles/Dilution
The number of mix cycles(0 to 99) per dilution. A zero(0) value will eliminate the mix
function.
Aspirate Speed
See Aspirate Speed in the Aspirate.
Dispense Speed
See Dispense Speed in the Dispense Function.
Row/Column Offset
The location for Dilution 1. All subsequent dilutions will follow left to right or top to
bottom of the plate, one row at a time. See Quad & Row/Column Offset in Stage
Operations for more details.
Ripple Volumes
This is a convenience when setting up the serial dilution to allow the same volume to
automatically be set for multiple successive dilutions. For example, if Ripple
Volumes is checked and I change Dil. 2 Mix volume, Dil. 3 Mix volume and
down will automatically be set to the same volume. Dil. 1 Mix Volume would be
unaffected. Ripple Volumes is a design time tool only. It has no bearing when the
program is actually running.
Dilution # Mix
The mix volume for each dilution performed.
Dilution # Xfer Out
The transfer volume from current dilution to the next. If a transfer out volume is
specified for the last dilution, that volume will be in the tips at the completion of the
function.
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MICRO-LOGGING:
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3.27 Shaker Nest
A stacker accessible nest that has an electronic oscillating mechanism that
can be turned on and off programmatically in the Quadra. The oscillation
amplitude is controlled by a dial on the Shaker Nest.
Conditions
You must have a shaker nest on the specified shuttle position, properly connected
to the Quadra in order for this function to work.
Remarks
Important Note: Powered accessories such as Vacuum Traps, Autotraps and
Shaker Nests, should not be connected with the Quadra 4 powered on. Please
turn off AC power BEFORE connecting these accessories.
Specify which shaker nest to control, and whether or not to switch the device on or off.
3.28 Shuck Tips
Shuck Tips removes disposable tips from the tip pins or removes fixed tip assembly from
the tip head.
Conditions
To Shuck Tips, select a shuttle position that has a tip capable fixture.
Remarks
Shuck Tips
Sets the function to shuck tips from the selected shuttle position.
Cycle Pinch Tubes
When checked, the pinch tube will open before the tips are shucked, and close
after the stage is lowered. This feature is available on pumps with pinch tubes.
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3.29 Shuttle Layout Change
Shuttle Layout Change allows the user to pause the program that is running,
and change the fixtures and components loaded onto the shuttle.
Conditions
There are no requirements for this function.
Remarks
Clear Shuttle
Layout
Clear Shuttle Layout gives the user an empty shuttle to work with when changing the
fixtures and components.
Import Previous Shuttle Layout
Import Previous Shuttle Layout brings all of the hardware previously installed on the
shuttle, and allows the user to add/change the existing fixtures and components.
Pause On Layout Change
Pause On Layout Change will pause the running program, and post the message typed
into the Edit Box below. The user has an infinite amount of time to change the layout
of the shuttle. The program will not resume until the user clicks the OK Button.
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3.30 SPE Vacuum Box Operation
Program control for the SPE Vacuum box.
Conditions
The SPE Vacuum Box operation requires a vacuum box and a vacuum controlled
Mini-Trap.
Remarks
Important Note: Powered
accessories such as Vacuum
Traps, Autotraps and Shaker
Nests, should not be connected
with the Quadra 4 powered on.
Please turn off AC power
BEFORE connecting these
accessories.
Load Plate
Message
When checked, a custom message will be displayed to instruct the operator. If not
checked, a generic message will be displayed.
Cycles
The function can have up to four vacuum cycles. Each cycle will set the inches (0>25) of vacuum to use and the duration. When the "Wait for Vacuum" box is
checked, the function will delay until the vacuum reaches the desired setting.
Remove Plate Message
When checked, a custom message will be displayed to instruct the operator. If not
checked, a generic message will be displayed.
Note: This operation requires that special hardware be installed.
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3.31 SPE Pressure Vacuum Operation
The SPE Pressure/Vacuum function allows for both positive pressure and vacuum to be
applied to a filter plate.
Conditions
This function requires that the vacuum controlled mini-trap and the software controlled
pressure regulator options be installed.
Remarks
Plate Load Message
Message to prompt the operator to prepare for this function.
Stage Height
Height to move the vacuum box/plate to the pump pressure fixture. Note that the stage
will force the vacuum box/plate into the pressure fixture, because of this the STAGE
SAFETY is bypassed.
Cycles
The function can do up to four pressure/vacuum cycles. The cycle can operate both
vacuum and pressure or either. To turn off either set the value to zero.
Clear Plate on Exit
When checked, a puff of air will be cycled to free the plate from the pump pressure
fixture.
Vacuum Pressure Relief Time
Time required to bleed off the vacuum/pressure to free the plate from the vacuum box.
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3.32 Stacker Operations
The Stacker Operations Function allows the user to Infeed and Outfeed plates from the
stackers at various times throughout the length of the program.
Conditions
The Stacker Operations Function requires you to select a shuttle position that has a
plate capable fixture. The fixture also has to allow stacker related functions.
Remarks
Select Stacker
Select the stacker you would like to Infeed from or Outfeed to.
Infeed or Outfeed Shuttle Position
Select you appropriate operation, whether you are bringing plates onto the shuttle
(Infeed), or returning them to their original, or different stacker (Outfeed).
Disable Lid Capability indicates that if the stacker supports lids than the
plate being processed does not have one. This is necessary for thin plates.
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3.33 Target Method
The "Target Method" function is available for the 450ul Timed Dispense head and
the 20/50/250ul DMV head.
The function is a tightly coupled Aspirate and Dispense of small volumes.
Remarks
TARGET(SOURCE) - The "Target" volume. height, speed, etc. Set the "Source"
plate volume. The operation will pre-wet the tips, aspirate the default tip volume,
and then dispense the delta to achieve the target volume.
TARGET(DESTINATION) - The "Dispense" height, speed, timed dispense height, etc.
Set the "Destination" plate. Generally all but 3ul is dispensed at the destination
plate. The remainder is dispatched by a timed dispense function. If the volume is
less than 3ul, the dispense is skipped, and the timed dispense will be done.
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NOTES: TARGET METHOD PIPETTING
The target method takes advantage of the fact that higher positive pressures
may be developed within the tip on a dispense motion, compared to the low
negative pressures that are available for aspiration. The target method is only available
for pipetting volumes less than 50µL. If target method is selected on the source
characteristics of any template, it will automatically aspirate and dispense 30µL, first
to pre-wet the tip. Then it will aspirate a 50µL volume and automatically
dispense back to the target volume to be dispensed. For example, assume 10µL is
the desired target volume. 50µL is aspirated and 40µL is automatically dispensed back
to the source with the tip submerged in the source liquid.
If target injection is selected for the destination characteristics, then the following action
will occur if the delivery volume is less than 50µL. The dispense will automatically be
made at the top of the well (as determined by the database for the plate selected in the
deck layout. You must use the correct plate layout). All but 3µL of the tip volume will be
dispensed by piston displacement.
The final 3µL will be blown out using the time dispense function. The air pressure for the
time dispense function should be set at 3 psi.
The 3µL blow out volume was derived empirically. At 8µL, the fluid flow was not
fast enough to overcome the surface tension forces at the orifice and the dispensed
liquid may form a drop on the tip exterior. The small 3µL volume and 3 psi driving force,
accelerates the liquid fast enough to minimize or eliminate the clinging drops on the tip
exterior.
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3.34 Timed Dispense
To deliver air or reagent from the Internal Air Reservoir
Conditions
To Time Dispense you will need to select a shuttle position with a plate capable
fixture.
Remarks
Cycles
The number of times the pinch tube clamp opens to deliver air.
Height
The height which the tips will deliver air to the well.
Note: Check air pressure to avoid high pressure dispenses that spill liquid
(this can be done from the Utility).
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3.35 Vacuum Box Operation
The vacuum box operation is used to control the automated vacuum box.
Conditions
None
Remarks
Open Vacuum Box performs vacuum off, tray down, open box, and then tray
up. If Run Raw is checked then the box will just open.
Close Vacuum Box performs vacuum off, tray down, and then close box. If Run Raw
is checked then the box will just close.
Turn On Vacuum performs vacuum off and then raises or lowers the tray (as indicated
by the checkbox) before turning on the vacuum. If Run Raw is checked then the
vacuum will just turn on.
Turn Off Vacuum performs vacuum off and then raises or lowers the tray (as
indicated by the checkbox). If Run Raw is checked then the vacuum will just turn off.
Raise Tray performs vacuum off and then raises the tray. If Run Raw is checked then
the tray will just raise (Warning: It is not recommended to raise the tray while vacuum
is on).
Lower Tray performs vacuum off and then lowers the tray. If Run Raw is checked
then the tray will just lower (Warning: It is not recommended to lower the tray while
vacuum is on).
Run Raw when check will circumvent implied action for the selected operation (not
recommended). See each operation for the effect of checking this option.
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3.36 Vacuum Control
This function allows the user to set the trap vacuum.
Conditions
The software controlled Mini-Trap feature must be installed.
Remarks
Set Vacuum - Sets the desired vacuum to 0 to 25 inches of mercury. Note that a 0
setting turns the vacuum off.
Set Vacuum and Wait/No Wait - When the "wait" feature is selected, the program will
pause until the trap reaches the desired vacuum. This usually takes from 10 to 20
seconds, depending on the vacuum source. when the "No Wait" feature is selected,
the trap vacuum is set, but the program will not pause.
Important Note:
Powered accessories such as Vacuum Traps, Autotraps and Shaker
Nests, should not be connected with the Quadra 4 powered on.
Please turn off AC power BEFORE connecting these accessories.
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3.37 Wash
A series of alternating aspirate and dispense steps to cleanse the tips.
Conditions
This function requires Ultrasonic Wash to be on the specified shuttle position in
order for this function to operate properly.
Remarks
Blow Out Volume
Blow Out Height
Mix/Wash Volume
See Mix Function.
Mix/Wash Cycles
See Mix Function.
Aspirate Speed
Aspirate Height
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Dispense Speed
Dispense Height
Wash Delay Timers
Pre-wash time and post-wash time are used with flow wash. Pre-wash time is the
number of seconds to delay before pipetting after water flow has started. Post-wash
time is the number of seconds to delay before turning off the water flow after
pipetting.
Dry/Rinse Tips
The Dry/Rinse Tips option allows the user to blow out an air gap at the end of the
wash.
Timed Dispense
See Timed Dispense Function.
Drop Stage after Tip Wash
See Additional Options.
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3.38 Program Operations Common Options
3.38.1 Additional Options
Slow Tip Withdraw
This options will extract the tip from the liquid at a slower speed to help reduce residual
liquid on the exterior of the tip. The tip withdraw speed can be set from 1 to 4, a speed
of 0 turns off the slow withdraw function.
Drop Stage After ...
This option brings the stage to a zero height after the function has concluded.
Disable Bar Code Scan
If the bar code scanner is enabled in the logging options section, this option allows the
user to disable reading the bar code for the current step.
Enable Liquid Following
Allows the user to minimize tip submersion during an aspirate or dispense function.
Once the initial aspirate or dispense height is programmed, the software automatically
adjusts the tip position in a well deeper or higher, respectively, as the liquid level in a
reservoir or micro-plates changes. This rate of adjustment is based on the Well Rate
parameters assigned in the micro-plate and reservoir database.
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3.38.2 Dip
Tips
The Dip Tips operation is available for the Dispense, Mix, and Empty Sample
Functions.
Conditions
The operation is used to remove excess liquid from the tips after executing the
function.
Remarks
The Dip Tip setting is a Delta from the Dispense/Blow Out/Empty height. For
example if the dispense height was set to 1000, the stage will move 19 steps to
1019 and then back to 1000.
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3.38.3 Select Shuttle Position
This function allows the user to change the shuttle position of a step without having to
recreate the step. Grayed positions are unavailable for the current function.
3.38.4 Stage Operations
You have two options to configure you stage for special plates. One is Quadrant
& Row/Column Offsets, and the other is X-Y Raw Stage Offsets. These functions
allow you to take a 96 tip head, and pipette to a 384-Well plate.
Quad & Row/Column Offsets
Note, since the Quadra 4 SPE does not have an indexing stage, the Stage Operations
selection is locked out.
Using the Quadrant Formatter feature, it is easy to take a standard 96 Tip head, and
format it to pipette from a 384-Well Plate.
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Simply by selecting what particular quadrant you are aspirating from, the application can
locate the proper well.
Along with quadrant reformatting, you can also specify Row and Column Offsets.
What this function does is allow the user to move the plate along the X and Y axis
in order to line up the tips with another particular well. Offset values represent tip
position relative to standard labeling for plates with the same density of wells as tips.
Row Offset will position the tips one row up or down if the value is negative or positive
respectively. Column Offset will position the tips one column to the right or left if the
value is positive or negative respectively.
The following are a couple of examples that may help you understand this concept
more:
Assuming you have a 96 Well Plate, and one tip inserted into well E6...
1. If you want to pipette from Well E7, you would set your column offset to +1.
2. If you want to pipette from Well E5, you would set your column offset to -1.
3. If you want to pipette from Well D6, you would set your row offset to -1.
4. If you want to pipette from Well F6, you would set your row offset to +1.
5. If you want to pipette from Well F7, you would set your row offset to +1, and your
column offset to +1.
6. If you want to pipette from Well D5, you would set your row offset to -1, and your
column offset to -1.
When using higher density plates with a lower density tip head, such as a 384 well plate
and a 96 tip head , Quadrant Formatter and Row\Column Offsets must be used in
conjunction with each other to reach the desired well.
Assuming you have a 384 Well plate & a 96 Tip head, and one tip centered between
wells A1, A2, B1, & B2...
1. If you want to pipette from well B3, you would set the Column Offset to +1, and
quadrant to 4.
2. 3.
X-Y Stage Raw Offsets
Touch Off
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Using the Raw setting allows the user to manually enter the number (of steps) that the
stage has to move in either direction in order to get at the well you need to. Values
represent tip movement in a Cartesian plane (although the stage actually moves.) In a
standard 96-Well plate, there are 577 steps between each well, and in a standard 384Well plate, there are 288 steps between
each well. Here are some more examples:
Assuming you have a 96 Well Plate, and one tip inserted into well E6...
1. If you want to pipette from Well E7, you would set your X Offset to +577.
2. If you want to pipette from Well F6, you would set your Y Offset to -577.
Assuming you have a 384 Well Plate, and one tip inserted into well C3...
1. If you want to pipette from Well C4, you would set your X Offset to +288.
2. If you want to pipette from Well D3, you would set your Y Offset to -288.
3.38.5 Touch Off
The side touch off is used to remove excess liquid from the tips.
Conditions
This option is selectable in the Dispense, Mix, and Empty Sample functions.
Remarks
The multi-point side touch off is available for full x-y indexing stages. The X touch off is
the standard single point movement. The XY, and XY -X -Y touch offs are 2 and 4 point
movements.
All touch off movements start from the center of the well to the offset and back to the
center for each point.
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4.0 Database Editor
Getting Started with DBEditor
The Database Editor allows you to manipulate the values for specific plates and
fixtures easily and reliably. You can start the Database Editor by select the
Database Menu > Edit Plate, Reservoir, and Fixture Database.
The left side of this form displays a tree for Plates, Reservoirs, and Fixtures. Under
each primary branch, there is the first level of categories for each item. Plates are
separated by the number of wells, and Fixtures are separated by there basic
functionality.
The Menu Bar
The Plate Panel
The Fixture Panel
Importing Plates-Reservoirs-Fixtures
When pipetting liquids, an important parameter is the location of the pipettor tip within
the well. This is accomplished with the use of the plate and reservoir database on all of
Tomtec’s Quadra’s.
When aspirating liquid, the tips should be near the bottom of the well, but not pinned.
When dispensing liquid, there are two basic choices. If it is desired to mix the
dispensed liquid with the contents of the well, then the dispense should be made at the
bottom of the well. Again, near the physical bottom, but not pinned. If a non contact
dispense is to be made, then the tips must be at the top of the well, but within the
confines of the top rim.
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Tomtec’s Quadra design allows another important pipetting feature. It provides the
ability to blow low air pressure through the tip orifices. This is referred to as the “Timed
Dispense Function”. The low air pressure setting is controlled by a precision pressure
regulator. Normally, it is set from 0.5psi to 1.0psi. This surge of positive air pressure is
used to clear the residual that is retained within the tip orifice, due to capillary action. It
is of significant importance for pipetting accuracy at small volumes.
The timed dispense function may also be used for true non-contact dispensing. For that
application, all but the last 3µL of a dispense volume is dispensed using piston
displacement. Then this last 3µL is cleared with the “Timed Dispense Function”. The
3µL slug of liquid normally has sufficient kinetic energy to clear the orifice without
leaving clinging drops.
Where should the “Timed Dispense Function” be made? Normally the tips should be
at the top of the well, but within the confines of top rim. However, if the well volume is
near the top, there is concern for splashing the well contents.
Tomtec’s software provides a third tip height in relation to the well. This is “Clear
Height”. This is defined when the tips are .060 inches above the top of the wells. This
height is used if the stage must index the plate under the tips, for serial dilution as an
example.
In summary, the Quadra software allows the user to set very specific tip heights.
However, this is too tedious and not required for normal use. It may be of value on a
specific protocol. For normal use, the Quadra database software provides 3 specific tip
locations for the specific plate or reservoir that has been selected. They are as follows.
Clear Height:
The tips will clear the top of the plate by .060 inches.
Top of Well:
The tips are located .060 inches below the top rim of the
well.
Bottom of the Well:
The tips are .030 inches from touching the bottom of the
well.
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4.1 The Menu Bar
The File Menu
The File Menu consists of two main functions: Importing records from another file, and
exiting the program.
From the File Menu you can Import Plates, Reservoirs, and Fixtures. When
selecting the appropriate category, a dialog box appears.
This Dialog box will allow you to import from database files stored in the Quadra 3 2.6.x
format (*.txt) and the new 3.0.x format (*.dat).
Upon selecting a file, the Import Window will appear. Please refer to the section on
Importing Plates and Fixtures for a detailed explanation of the importing process.
The Edit Menu
The Edit Menu allows you to add new records, Copy items, delete items, and also open
the Quadra Utility screen when required.
The View Menu
The View Menu gives you the option for customizing the view of the screen by
either including the small images next to their relative item, or negating them.
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4.2 The Plate Panel
When setting the parameters for a plate or reservoir, the Plate Panel will appear on the
right hand side of the form. This form will allow the user to add/change the plate name,
Clear Steps, Top Steps, Bottom Steps, the Volume, the touch off steps, and also
calculate the well-rate, a feature that controls the liquid follow function.
Clear Top Steps
Clear Top Steps is the number of steps the stage has to travel beyond the fixture in
order to clear the plate/reservoir entirely for serial dilution or quadrant moves.
Top Steps
Top Steps is the number of steps the stage has to travel beyond the fixture in order to
set f the tips at the top of the plate.
Side Touch Off Steps
Side Touch Off Steps is the number of steps the stage indexes in the horizontal
direction, in order for the side of the tips to touch the side of the well.
Bottom Steps
Bottom Steps is the number of steps the stage has to travel beyond the fixture
in order for the tips to touch the bottom of the wells. (Not necessarily the bottom
of the plate or reservoir.)
Volume
Volume is the amount of liquid (in µL for plates and mL for reservoirs) that a single well
or reservoir can hold.
Well Rate
Once Top Steps, Bottom Steps, and Volume are entered, it is possible to estimate the
well rate by clicking on the button next to the field. This field is user editable, and it is
advised that you test, and calibrate the well rate manually. The application will only
provide an estimate.
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4.3 The Fixture Panel
Fixtures are easier than plates to calibrate because there is only one setting,
the Calibration Height. This is, by definition, the height at which the tips meet the
fixture, without pinning the fixture to the stage.
There are several fixtures, preinstalled into the software, that are not usereditable. The only attribute the user may edit is the calibration height. Any new
fixtures, or custom fixtures are user editable, and must specify whether or not they can
operate with a Quadra Stacker, whether or not they can operate with a Tip Feeder, and
whether they can be pipette from/to.
Tip jigs are fixtures that require no user intervention. All tip jigs have a load height of
zero. If any value is inserted into this field, it will override the load height set in the
system parameters function. The same is true for the shuck height.
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After setting the Load Height, if you are unsatisfied with the result, you may
click the Reset Calibration button to return to the original value, only before you
save. The application will prompt you to save any changes when you switch records, or
close the application.
A new category has been added, called, "Liquid Containers." These are fixtures
that contain calibration heights as well as top steps so when programming a
function, the dialog can properly represent tip position inside the container.
4.4 The Touch Off Panel
The Touch Off Panel is a special feature that facilitates the accurate depicting of the
value for the Side Touch Off Step control.
The process is as follows:
1. Open up the Quadra Utility, and position the tips in the center of the designated well,
with the tips just barely inside the well.
2. Close the Quadra Utility.
3. Enter the number of steps for Side Touch Off.
4. Click Test Touch Off to move the tips.
5. The tips will hold position until the user clicks yes or no to accept the value.
6. Click No and Repeat steps 4-6 until value is accurate.
7. Click yes to accept the value and populate the Side Touch Off Steps field.
4.5 Importing Plates-Reservoirs-Fixtures
Importing Plates/Fixtures is Quick and Easy. Simply click on the File Menu and
select Import Plates/Reservoirs/Fixtures.
Navigate to the proper folder and click Open to open that particular file.
A dialog box will automatically appear. The items on the left are those records
that are available in the selected file, and the items on the right are those you wish
to import. Click on the Add and Remove buttons to carry records from one side to
another.
When you are finished selecting those items you wish to import, click on the Import
button to finalize the procedure.
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5.0 Options
User Preferences
Default Plates
Default Reservoirs
Displayed Fixtures
Configure Model
Configure Com Port
Current File Settings
Set Arm Position Names and Heights
5.1 User Preferences
Auto - Connect/Initialize
The operator can set the application to:
1. Manually connect and Initialize the Quadra
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2. Connect to and then initialize the Quadra when the application starts.
3. Auto-Initialize the Quadra after connecting.
Auto - Disconnect On Application Exit
When disabled, the operator has to use the "Disconnect" tool bar or menu
item to shut down the Quadra. When enabled, the application will automatically call
the "Disconnect" function when the application is exited.
Simulation Mode
The application can be set to either run as simulation on the screen only or run on the
Quadra.
Prompt For Shuttle Position Data
When disabled, the operator has to "right" click on the shuttle position to add any
additional data about the position. If enabled, the operator will be prompted for
additional data every time there is a change.
Log File Directory
Designates where the log files are created and saved.
Recent File History Size
Sets the number(0 - 4) of program files to save to the recent file history list.
Tip Depth Trackbar
When enabled, a track bar is displayed between the edit box and the tip in well graphic
for editing stage heights. This provides an additional tool for scrolling quickly through
the stage heights. Without the track bar, scrolling can be accomplished by directly
scrolling the tip in the well or using the up/down buttons in the edit box.
System Defaults
The default slow tip withdraw speed, 1 to 4, for all functions that have this option. Note
that a speed of 0(zero) will turn off this option.
When checked, the default mode for shucking tips on pinch tube equipped pumps will
be to open the pinch tubes before shucking, and closing after. If the "DMV" head is
selected, the default tip volume can be set to 20, 50, or 250 µL. If the "Target Method"
function is checked, the default target volume can be set.
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Power Off PC On
Application Exit
When enabled and the UPS software is installed and configured, the PC will shut down
the PC/Laptop when you exit the application.
Auto-Pinch Tubes Open On Disconnect
This option is enabled by default every time a pinch tube equipped head is selected.
When enabled it will open the pinch tubes after a ten second count down:
The operator has the option to abort the operation anytime during the countdown.
When this option is disabled, the operator will be prompted to open the pinch tubes.
5.2 Default Plates
96 Well Plates
When this button is pressed, all 96 well plates that are in the database will be
displayed.
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384 Well Plates
When this button is pressed, at 384 well plates that are in the database will be
displayed.
1536 Well Plates
When this button is pressed, all 1536 well plates that are in the database will be
displayed.
Other Plates
When this button is pressed, all non-standard plates that are in the database will
be displayed.
All Plates
When this button is pressed, all plates that are in the database will be displayed.
Add
When this button is pressed, the selected plate will be moved to the default list.
Delete
When this button is pressed, the selected plate in the default list (lower dialog
box) will be removed.
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5.3 Default Reservoirs
Add
When this button is pressed, the selected reservoir will be moved to the default list.
Delete
When this button is pressed, the selected reservoir in the default list (lower dialog
box) will be removed.
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5.4 Configure Model
Select Quadra Configuration
The operator can set the model configuration by selecting the pump type.
Functions Palette
The operator can select or deselect which optional functions to display. Note that
only the functions buttons are affected. Editing of functions not displayed on the
buttons, but in the program is allowed.
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5.5 Configure Com Port
Com Port
The operator can set the serial com port(1 to 32), the baud rate, and flow control.
Check Quadra documentation for baud rates and flow control.
5.6 Set Arm Position Names and Heights
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This dialog allows the user to define commonly used heights for the arm. They will be
made available when setting up an Arm Plate Transfer Operation.
Calibrate becomes visible when enter a position number’s edit box. Selecting Calibrate
will bring up the Utility on the arm page.
Open the hand and position it as desired. Upon exiting the Utility dialog a prompt will
occur to automatically set the value to the current arm position.
Upon exiting the user defined arm heights dialog a prompt will occur to retract the arm if it
is extended.
5.7 View User Preferences
This feature will display the current user preference settings. The settings can be
printed or saved to a file.
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5.8 Displayed Fixtures
Configure Displayed Fixtures
This function allows the user to customize the Labware - Components group on the
application main window (see Edit Display).
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5.8.1 Edit Display and Order of Fixtures
Add
This function will add the selected fixture to the display list.
Remove
This function will remove the selected fixture from the display list.
Up
This function will move the selected fixture up in the display list order.
Down
This function will move the selected fixture down in the display list order.
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5.9 Current File Settings Options
Logging Options
Runtime Database Usage Options
5.9.1 Logging Options
Log Type Check Boxes
The log type is set by checking the box. The Standard Log defaults with the
Time/Date Stamp feature enabled. Bar Codes can be recorded in the Standard
Log, or in a Bar Code Only log. When the "Log Single/Block Steps" is checked,
partial program runs are also logged.
Log Name
The log generated can have the same name as the program, the operator
can be prompted for a log name at the start of the program, and the log file
name can be generated automatically based on the program name, time and date.
Select Bar Code Reader Mode
When a bar code type log is selected, the "Select Bar Code Reader Mode"
group is enabled. In this group the operator can select when bar codes should
be scanned. In an Aspirate function, for example, " UP" would cause the bar code
scan to occur while the stage is raising the plate, before aspirating.
Bar Code Error Handling
"NO READ Errors" occur when the reader is unable to properly scan the bar code
on the applicable plate. Some conditions that would generate this error include a
missing bar code label or bar code label is on the wrong side.
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"BAD DATA Errors" occur when the Quadra is unable to properly communicate
with the reader. Typically this is caused when the BCR jumper cable is either not
connected to bar code reader ports or a connected cable has the incorrect wiring.
The Bar Code Error Handling group is enabled whenever a bar code logging is
required. The operator has the option of ignoring an error, recording the error
without intervention, or be prompted to enter the proper bar codes on an error.
Set as Default
The current logging options and runtime database usage settings
are saved as default settings for new programs.
Reset to Default
The current logging options and runtime database usage settings
are reset to default settings for new programs.
5.9.2 Runtime Database Usage Options
When this feature is selected, the plate and fixture heights are ignored, and raw
stage heights are used. The default is to use the plate and fixture database
heights, which requires plates and fixtures present at runtime. You might want to
check this if you are moving a program to another system where plates and
fixtures may not be in the database.
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6.0 Utility
Shuttle/Stacker
Stage
Pump
System
Backfill
Trap Operations
Miscellaneous
Arm - Vacuum Box - Aux. Station
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6.1 Shuttle/Stacker
Set Shuttle Position
Select the button to move to the desired shuttle position. Initialize Shuttle will
reset the shuttle and move to position 5. This would be useful in the event that a
shuttle position error has occurred.
Stacker
Outfeed will send a plate from the nest into the selected stacker.
Infeed will send a plate to the nest from the selected stacker. Stackers that are ‘lid
capable’ will have an additional push button that when down will disable the lid
handling capability. This is necessary for thin plates that do not have lids.
Initialize Stackers will reset all stacker elevators to the down position. This
would be useful in the event of a stacker lost or interlock error has occurred.
Tip Feeder
Outfeed will send a set of tips plate from the tip jig into the selected tip feeder.
Infeed will send a set of tips to the tip jig from the selected tip feeder.
Initialize Tip Feeders will reset all tip feeders to the up position.
Note: Tip Feeder operation will only be available if the device is present.
De-Lidder
Up Arrow will take the lid off the plate currently under the de-lidder.
Down Arrow will put the lid on the plate currently under the de-lidder.
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6.2 Stage
Set Stage Height
Move Stage will move the stage to the specified height in the adjacent edit box.
Holding down the shift key while selecting Move Stage will move stage to the 0
position.
System Safe Height will move the stage to height where the stage can
safely clear the shuttle in both the x and y direction. Warning: This height will
be sufficient to index most nest/plate combinations, but certain deep well plates
must be indexed below the safe height to prevent tip-well collision.
Jog Up will move the stage up 20 steps.
Jog Down will move the stage down 20 steps.
Single Red Up Arrow will move the stage up 1 step.
Single Red Down Arrow will move the stage down 1 step.
Initialize Stage will reset the stage top at the center position and then move the
stage down to the zero position. This would be useful in the event a stage position
or safety error has occurred.
Note: If a stage safety occurred, the stage override UP/DOWN can be enabled by
holding down the "Alt" key when clicking on the stage up/down arrows. The Stage
MUST be reinitialized after clearing the safety error.
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Set Stage - XY Offset
Index - X will index the stage as specified in the adjacent edit box. Value represents tip
movement in a Cartesian plane (although the stage actually moves). Holding down
the shift key while selecting Index – X will move the X portion of the stage to
the 0 offset position.
Index - Y will index the stage as specified in the adjacent edit box. Value represents tip
movement in a Cartesian plane (although the stage actually moves). Holding down
the shift key while selecting Index – Y will move the Y portion of the stage to
the 0 offset position.
Single Red Up Arrow will move the stage 50 steps toward the back of the Quadra.
Single Red Down Arrow will move the stage 50 steps toward the front of the Quadra.
Single Red Left Arrow will move the stage 50 steps left relative to one facing the front of
the Quadra.
Single Red Right Arrow will move the stage 50 steps right relative to one facing the front
of the Quadra.
Home Stage - XY will reset the XY position to the center of the stage. This is useful in the
event that the XY portion of the stage has lost its true position, perhaps due to a physical
collision.
Stage Formatting
Select the button to move to the desired Quadrant.
WARNING: Stage indexing will occur at its current height. Make sure that the plate
has sufficient clearance of the tips and the stage is not completely below the
shuttle.
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6.3 Pinch Tubes
Timed Dispense
Timed Dispense will dispense air or reagent for the specified
number of cycles in the adjacent edit box. Note: the amount of
volume dispensed per cycle is controlled by the pressure in the system air
tank.
Set Pinch Tubes
Open Pinch Tubes will open the inch tubes.
Close Pinch Tubes will close the pinch tubes.
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Pressure Valve
The pressure valve can be closed to keep the compressor from constantly running.
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6.4 Pump
Set Pump
Aspirate will aspirate the volume (in µL Liters) specified in the adjacent edit box.
Dispense will dispense the volume (µL Liters) specified in the adjacent edit box.
Empty will dispense all volume in the tips.
Initialize Pump will reset all tips to the zero volume position.
Volume
Displays or sets the volume to aspirate or dispense. The value(s) in the brackets are
the current pump volume and the expected pump volume. If both values are the
same, only one value will be shown, but if the values are different both will
be displayed.
The values above are showing the actual pump volume (8.5µL) verses the expected
pump volume (10.0µL).
Set Tips
Load Tips will load tips onto the pump from the current shuttle position. When a
"DMV" head is detected, the tip volume can be set. The tip volume limits the aspirate
and dispense function's maximum volume.
Shuck Tips will shuck tips from the pump to the current shuttle position. Make sure
the appropriate tip jig is present.
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6.5 System
Set System
Connect will start communication with the Quadra and set the Quadra under
remote control. Connect is required before any other action.
Disconnect will end communication with the Quadra and return the Quadra back to
local control. Initialize System will reset all devices on the Quadra to their default
positions.
6.6 Trap Operations
Standard Trap:
Trap Control
Refresh Status will update the adjacent progress bar to indicate the current level of fluid in
the flow wash trap.
Empty Trap will empty the trap
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Filter Control
Start Filtering will turn on the vacuum to the auto-vacuum box.
Stop Filtering will turn off the vacuum to the auto-vacuum box.
Mini-Trap:
Vacuum Control
Set Vacuum will set the vacuum in inches.
Get Vacuum displays the current vacuum setting in inches.
Refresh Vacuum does a quick vacuum set to the value displayed in the edit box.
Vac Off turns off the vacuum to the vacuum box.
Vac On turn on the vacuum to the vacuum box.
Important Note:
Powered accessories such as Vacuum Traps, Autotraps and Shaker
Nests, should not be connected with the Quadra 4 powered on.
Please turn off AC power BEFORE connecting these accessories.
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6.7 Miscellaneous
Magnet Device
On/Off is used to turn the magnet on or off for the selected Magnet Nest.
Shaker Device
On/Off is used to turn shaking on or off for the selected Shaker Nest.
Set Air Pressure
Start Monitoring will continuously monitor and display the air pressure. The
button text will change to Stop Monitoring once selected. Selecting the button
then will stop the continuous monitoring.
Note: The air pressure is changed by turning the black gauge knob on the
side of the Quadra.
Important Note:
Powered accessories such as Vacuum Trap and Shaker Nests, should
not be connected with the Quadra 4 powered on. Please turn off AC
power BEFORE connecting these accessories.
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6.8 Arm - Vacuum Box - Aux. Station
Set Arm
Get Plate will open hand, move arm to position specified in the edit box,
close hand, and then retract the arm.
Put Plate will move arm to position specified in the edit box, open hand,
and then retract the arm.
Move Arm will move the arm to the position specified only. The hand will not
be opened or closed.
Safe Height will move the arm to the defined safe height.
Jog Up/Jog Down will move the arm up or down in either single or 20 step increments.
Open Hand will open the hand at its current position.
Close Hand will close the hand at its current position.
Initialize Arm will reset the arm.
Set Vacuum Box
Vacuum On/Off turns the vacuum on or off.
Open/Close Box opens or closes the box top.
Tray Up/Down raises or lowers the tray. Make sure the vacuum is off first.
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Set Aux Station
Down Arrow (1) moves the auxiliary station to position 1 under the arm.
Up Arrow (2) moves the auxiliary station to position 2 under the arm.
6.9 Status
The light switch status utility gives the user a real time assessment of the Quadra 4's
inputs.
When depressed, the status of the pump, stage, shuttle, stackers, trap, arm, interlocks,
and auxiliary inputs are displayed.
6.10 Pump Calibration Tables
The pump calibration tables are supplied by Tomtec to insure the accuracy of the
pump over its range. The curve(s) supplied are in a special ".csv" format. The data
entry application reads in the supplied data, displays the curve, and saves it in the
"QCURVEDB.DAT" file.
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Remarks
Select Liquid Class
Any current liquid class can be selected and displayed along with its curve.
The most recent curve for the pump type is always displayed.
The data displayed in the table can be edited or deleted by right clicking on the
desired line.
Import Curve
The "csv" formatted curve supplied by Tomtec can be selected, read in and
displayed:
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The file is checked against any duplicate named curves. If a duplicate is found,
the operation will be prompted:
A "Yes" reply will add " - Ver: xx" after the curve name.
New
A new curve can be named and entered into the database.
If the model uses a DMV head, the tip volume has to be selected. This allows for a
proper curve display.
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Upon clicking on the OK button, the data entry function will be displayed.
When the OK key is clicked, the data table will fill, and the new curve will by plotted.
Delete
The currently displayed curve will be deleted from the database.
Note:
Aspirate, Dispense, and Target Method functions can use the curve data. When
curve data is used, the pump is commanded to move in steps, not by volume. The
number of steps to move the pump is calculated as follows:
Exact Match:
diffvol = requestedvol - measuredvol
xx= (diffvol * stepsperul)/1000.0
yy= (requestedvol * stepsperul)/1000.0
steps= yy + xx
The above data is "Micro Logged" as:
CalX: Vol: 10.0, Data: Req = 10.0 Meas = 7.7590, VDiff: 2.2410, SDiff: 35, Steps: Req
= 160 Act = 195
Interpolated Volume:
calvol = (requestedvol - measuredlow)/(measuredhi - measuredlow) * (requestedhi requestedlow) + requestedlow steps = (calvol * stepsperul) /1000.0
The above data is "Micro Logged" as:
CalI: Vol: 135.0, Data: ReqL: 125.0 MeasL: 125.8600 ReqH: 200.0 MeasH:
207.5100, CalVol: 133.39559094, Steps: 2134 = 133395.59094 * 16) / 1000.0
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7.0 Tools
7.1 Tools Serial Dilution Wizard
The Serial Dilution Wizard is a tool that will guide the user in setting up a serial dilution
protocol.
Conditions
The Serial Dilution Wizard requires a selected shuttle position.
Remarks
Setup
Set Source Row/Column
Set Sample Aspirate Volume and Height, Row, and Column Offsets
Set Initial Dilution Row/Column
Serial Dilution
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7.1.1 Setup
Sample Source Position
Select the source plate shuttle position.
Dilution Destination Position
Select the destination plate shuttle position.
Sample and Dilution Volumes
The Sample Volume is the initial volume aspirated from the Source plate and
transferred to the Destination plate to achieve a serial dilution.
The Dilution Volume is the pre-existing volume in each well of the Destination plate.
The Dilution Factor is calculated from the Sample and Dilution volumes.
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Serial Dilution Tip Arrangement
The tip arrangement group box allows for the selection of either a custom or preset tip
arrangement. The preset arrangements set the number of dilutions and initial row or
column offset. The custom tip arrangement allows the operator to set the number of
dilutions and initial row and column offset.
Next
The "Next >" button moves the wizard to the next page.
7.1.2 Set Source Row/Column
If the source plate is a 96 well plate, and a preset tip arrangement is selected, the row
or column for the initial Sample Volume aspirate can be selected by clicking on the plate
graphic.
Back
The "< Back" button moves the wizard back to the previous page.
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7.1.3 Set Sample Aspirate Volume and Height, Row, and
Column Offsets
See Aspirate Function for more details.
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7.1.4 Set Initial Dilution Row/Column
If the destination plate is a 96 well plate, and a preset tip arrangement is
selected, the row or column for the initial Dispense Volume can be selected
by clicking on the plate graphic.
7.1.5 Serial Dilution
See Serial Dilution Function for more details.
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8.0 Technical Tips
8.1 Introduction
Not only is it important to understand the function of the different transfer operations, it
is critical to arrange them correctly so that the protocol being designed functions
properly and can process the maximum number of plates in the shortest period of time.
To assure this, here are some suggestions for writing an effective protocol.
8.2 Pre-Planning
The first thing to do is determine what it is you want to accomplish, then write it down on
paper. Hypothetically, let us assume that the protocol to be created will go as follows:
Take 20uL of magnetic bead reagent (re-suspended) and add it to a compound plate
containing protein X. Mix the reagents, and then magnetize the compound plate for 2
minutes. Transfer the supernatant from the compound plate to an assay plate. Quickly
rinse the magnetic beads with 10-20uL ETOH. Remove the ETOH and dispose of it.
Repeat 20 times.
8.3 Labware
Now that you know what needs to be done, determine what you need to accomplish
with this procedure. In this case you would need: 20 assay plates, 20 compound plates,
a reservoir of magnetic bead reagent, a reservoir of ETOH, a waste station, a wash
station, a shaking stacker nest (optional), and a magnetic stacker nest.
8.4 Labware Location
Once you know what labware you need, you need to decide if you can fit it and where to
put it. First, unless it is removed, the ultra-sonic wash station should be located on the
edge of the shuttle (in order to accommodate the tubing), leaving you with 5 shuttle
positions to utilize. Use shuttle positions 4 or 6 because 1
and 3 can access the front & rear stackers so they are too valuable. Of the 40
micro-titer plates, only two are being used at the same time and therefore will only
occupy 2 shuttle positions.
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Because of their ability to infeed/outfeed to both stackers on a given side, positions
1,3 are best suited for plates being stacked. All 20 of each plate should be placed in
the rear stacker of the respective side. In this case the assay plates will be in the right
rear stacker and be infeed to position 3. The magnetic nest is going to be
used on the compound plate, so if the compound plates are on the left side, the
magnetic nest should be on position 1 (below the left rear stacker). Positions 2 and 5
are still open for the remaining reservoirs. The magnetic bead reservoir may be
placed anywhere in relation to the pipetting head and the compound plate (for
time conservation).
A logical choice would be position 2. The ETOH reservoir is also going to be used on
the compound plate, and therefore should be as close to the compound plate as
possible. In this case, position 6 would be the wisest choice. For a waste disposal
reservoir, the ultra-sonic unit is a practical choice because it can be emptied into a
vacuum trap and is already on the shuttle. However, since there is an empty shuttle
positions it is just as easy to use a separate waste reservoir located at positions 5.
In general, positions 2 and 5 are very important because they can access both sides
(left and right) of the robot. Position 2 is the MOST important because it can access all
4 stackers. While 1,2, and 3 can access both front and back stackers, 4, 5, and 6 can
only access the front.
8.5 Writing The Protocol
Now that the labware has been selected and placed in its proper location, the actual
writing can begin. Since all 20 plates are going to be processed in the exact same
way, the program should begin with a loop command x20. An assay plate and
a compound plate are both going to be brought in at the same time, so loop both
stackers x20. By selecting this option plates are infeed automatically to positions
1,3, and will be outfeed to the front stackers at the loop end command. The
transfer will now have both the "begin loop" command and the "end loop" command.
Leave these how they are for the time being. The next step is to aspirate the resuspended magnetic bead reagent.
To get the reagent re-suspended you can do one of two things, you can manually mix
it with the pipetting head a number of times, or you can chose to place it on
a shaking nest that will keep the reagent re-suspended. Assuming that the
more time effective shaker nest is used, the next step would be Aspirate 20uL
from position 2. The next step would be to Dispense 20uL to plate in position
3. The bead reagent now needs to be mixed with reagent X in the plate. Select
the mix option on the left of the screen. For more efficient mixing, aspirate off the
bottom of the well and dispense higher up in the well. The magnetic nest needs to be
turned on in position.
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1. Under the options icon you will find the magnetic separation device option. Select
this to turn on the magnets at position 1. At this time you would like the beads to
settle toward the bottom of the nest for a full two minutes. The
recommendation at this time would be to wash your tips in the ultrasonic
bath for two minutes or to wash the tips and add a timed pause for the
remainder of the time (if it takes :50 sec to wash the tips, set a timed pause
70 seconds.
2. To yield the full 120 second wait time). Another method would be to lock-out that
position for 120 seconds (located under options). After the 2-minute pause the
supernatant needs to be transferred to the assay plate.
To accomplish this Aspirate 20uL from position 1, (with the magnets still on)
Turn the magnets off, and then Dispense 20uL into assay plate in 4. At this time you
should wash the tips to avoid contaminating the other reagents. The final step is to rinse
the magnetic particles with ETOH. To do this, Aspirate 15uL from the ETOH reservoir in
6, Dispense 15uL into position 1, and Mix 12-15uL in 1 a few times. To avoid removing
the magnetic particles, turn the magnets back on. You may wish to wait a few moments
to allow the beads to travel to the bottom of the wells so a pause of a few seconds can
be added, if not Aspirate 15uL from position 1 and dispense it into the waste station at
position 5. The protocol has now finished 1 plate. You can’t forget that the next 19
plates must be done in the exact same way, so wash the tips one last time to
prepare them for the next go around. The final touch is to place the end loop in its’
proper location. To do this, go to the beginning of the transfer and select the second
step, the "end loop" command. Once this step is highlighted, go over to the
right side of the menu and select the move down icon. The step has now gone
from being step two to step three. Keep pressing the move down icon until the
end loop is the last step. The protocol is now completed and ready to run.
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8.6 System Parameters
The Quadra 4 firmware parameters are set by qualified Tomtec personnel during
assembly.
Conditions
A password is required to change all but a few non-critical system parameters.
If a critical parameter has to be changed, you should contact Tomtec's Technical
Support department at 203.281.6790.
The pictures shown do not represent actual instrument settings.
9.0 System Parameters - Password
The Quadra 4 system parameters are password protected, but you can view them
by clicking the "Cancel" button on the password prompt without a password entry.
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9.1 System Parameters - Stage
Quadra 4 Stage XYZ Parameters.
9.2 System Parameters - X-Shuttle
Quadra 4 X-Shuttle Parameters
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9.3 System Parameters - Board A Misc.
Quadra 4 board A miscellaneous parameters
9.4 System Parameters - Pump
Quadra 4 pump parameters
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9.5 System Parameters - Y-Shuttle
Quadra 4 Y-Shuttle parameters
9.6 System Parameters - System Info
System Firmware and Software Version levels, Board Check Sums, and Serial
Number data. Serial Number - The serial number is entered by Tomtec personnel
when setting up the instrument.
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9.7 System Parameters - Shuttle Alignment Utility
Shuttle X-Y alignment Utility.
Remarks
This utility requires that the mechanical alignment be completed. What is adjusted
with this utility are the nest pickup & drops.
The Shuttle Alignment Utility allows the user to set the stage/shuttle/pump
alignment. The "Adjust Shuttle" buttons will jog the shuttle by the number of
steps shown in the center button. Clicking on the center button will increase
the jog step rate. The Set/Reset buttons will save or recall the position data. The
test program will go through shuttle positions 1 to 6 continuously until aborted by the
stop button.
Note: that once the end positions, Front, Rear, Left and Right, are set. They
should be the same for all shuttle positions.
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Remarks
This utility requires that the mechanical alignment be completed. What is adjusted with
this utility are the nest pickup & drops.
The Shuttle Alignment Utility allows the user to set the stage/shuttle/pump
alignment. The "Adjust Shuttle" buttons will jog the shuttle by the number of
steps shown in the center button. Clicking on the center button will increase the
jog step rate. The Set/Reset buttons will save or recall the position data. The test
program will go through shuttle positions 1 to 6 continuously until aborted by the stop
button.
Note: that once the end positions, Front, Rear, Left and Right, are set. They should be
the same for all shuttle positions.
9.8 System Parameters - View System Parameters
This utility allows the user to view, print or save to a text file the system settings.
Note: the default parameter file (Q4SYSPARM.DAT) is displayed. User named
files can be displayed by using the "Open File" button.
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9.9 Vacuum Calibration
Automated Mini-Trap vacuum calibration utility.
Conditions
The vacuum controlled Mini-Trap feature must be installed.
Remarks
Calibrate starts the procedure which consists of up to five vacuum set passes.
Each pass starts with the vacuum first set to zero(0) and then to 0, 5, 10, 15,
20 & 25 inches. After all passes have completed, the return setting for 0, 5,
10, 15, 20 & 25 inches in each pass are averaged and then compared against the
ideal value. From this comparison a delta is derived for each setting. The delta is then
used whenever a vacuum control function or trap utility is used. Note that if the
returned value is within +/- 2 inches, the delta is set to 0.
Abort stops the calibration.
Passes - The number of 0 to 25 inch passes to perform. The default is 3.
Apply accepts the calibration values for inclusion into the configuration file.
OK records & saves the values for program use.
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Section IV
PIPETTOR HEADS
The Quadra 4 design will accommodate different pipettor heads for specific applications.
They are all interchangeable by the user. Each pipettor head is secured with 4 bolts. The
head plugs into the back plane for its electrical and software connections. Each head is
then manually aligned to the stage. This can be easily accomplished in less than 30
minutes, start to finish. Each pipettor head and its use is described individually, since
any pipettor head may be placed in your Quadra 4.
QUADRA 4 AIR DISPLACEMENT PIPETTOR
HEADS
1 .0
Quadra 450µL Disposable Tips
This is the primary pipettor head for the Quadra 4. It is the dominant pipettor for
general applications with a long history of reliability and dependable operation. It has
several primary advantages. First is the 450µL volume of the disposable tip. Secondly,
the tips can reach the bottom of the SBS deep-well microplates. The third advantage
is the ability to blow pressurized air through the tips, under software control. This
function permits the combination of aqueous and organic pipetting, without having to
change tips between each liquid. It also permits non- contact dispensing. Another
function is to clear the residual retained in the tip by capillary action, when pipetting
small volumes.
The pipettor pistons are driven by a software controlled stepper motor. The motor
requires 400 steps to make one revolution. Each revolution of the motor moves the
individual 96 pistons 0.0625 inches. This distance multiplied by the piston diameter
equates to software steps per microliter.
Each individual piston has a small hole through it. This small hole is connected with
silicon rubber tubing, through a motorized pinch clamp, and then to an internal tank
that is pressurized with regulated air pressure from 0 to 5 psi. When the pinch point is
closed, all pistons act as air displacement pipettors, when the pinch point opens, air
pressure can blow through the tip orifices. This offers a number of distinct operating
advantages to the Quadra 4 user.
1 .1
Organic to Aqueous Pipetting
To pipette organic liquids, it is first necessary to fill the tip head space with the organic
vapors to prevent dripping. To switch back to pipetting aqueous liquids, it is necessary
for competitive instruments to change tips, to provide accuracy. The Quadra 4
program simply opens the pinch valve to clear the organic vapors. Now aqueous
liquids may be pipetted with accuracy. This is a common application in many sample
preparation techniques, solid phase extraction, liquid/liquid, and protein precipitation.
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1.2
Non Contact Dispensing
The major volume, contained in the tips, is dispensed above the receiving well, by
piston displacement. The final 3-5µL is dispensed with a timed dispense using air
pressure. The tips do not touch the contents of the well. Mixing may also be
obtained with this method of dispensing. The remaining volume to be blown out
and the air pressure used are controlled to be below the splash point.
1.3
Clearing Tip Residual
It is standard pipetting practice to initially aspirate blow out air, to be used to chase
the final residual, in the tip that is retained by capillary action. Using air pressure is
far more effective. A timed dispense is made that provides higher air velocity, to
clear the tip of capillary residual.
2.0
Quadra 450µL Tip Design
The tips for the 450µL head (PN 196-205) are specifically designed with a standard
orifice of 0.036 inches. The tips have a long skinny portion to reach the bottom of a
deep well plate. The upper barrel portion provides the volume. The lower portion
contains 90µL. It may be used for pipeline pipetting. This permits pipetting several
reagents sequentially, using small (10-15µL) air gaps for separation. This combined
with the multiple stations on the Quadra 4 shuttle, means several reagents may be
added in one pass. The larger volume buffer, which is aspirated first, is used to
wash out the smaller volume of drug compound, improving overall precision.
The Quadra tips are retained in their tip rack, allowing them to be loaded and
removed as an integrated individual tips may also be removed for creating pipettor
patterns, since the pipettor head has 96 individual pipettor pistons. Tips may be
removed for 48 well pipetting or 24 well pipetting. With the indexing stage of the
Quadra 4, a single row of tips may be used in the 8 or 12 direction for serial dilution
applications.
2.1
Loading the 450µL Disposable Tips
Tip racks are loaded manually when using the 450µL air displacement pipettor
head. The following describes the tip load protocol on all Quadras using this tip
including the Quadra Plus, Quadra 4, Quadra 3, Quadra 96 Model 320 and the
Quadra Tower.
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The tips are pressed on to the 96 tip pins by the upward motion of the stage.
Each tip pin has an elastomer “O” ring that seals each disposable tip to the
stainless tip pins in the Quadra head. To keep the tip load force within its design
limit, the “O” rings require periodic lubrication. A pre-lubricated chamois pad,
within the tip shuck plate, lubricates the “O” rings each time the tips are shucked.
Periodically, this chamois pad must be re-lubricated with a technical grade
mineral oil.
These Quadra tips are loaded by position on the tip pins. When fully loaded,
there should be less than 1/16 inch (.062 inches) clearance between the top of
the disposable tip and the Quadra shuck plate. This can be determined after
loading a rack of 96 tips by using your finger to push the tip tight against the
shuck plate. It should not move more than .062 inches. If it does, the Tip Load
Switch must be adjusted. If the tip is loaded tightly against the shuck plate, this
also indicates the need to adjust the Tip Load Switch.
The software will load the tips on command. If the instrument is unable to load
the tips on the first attempt, it will re-try 2 more times before displaying a “Tip
Load Error”. If the tip load switch does not break the contract, the tip load
function fails.
If there is a problem loading tips, it is typically due to one of two causes:
1. The primary cause of the tip load failure can be loss of lubrication on the tip pin
“O” rings. Use the tip pin lubricator that was supplied with the Quadra
(TOMTEC P/N 196-107).
2. The tip load switch is not set properly. Check the clearance between the top of
the tip and the tip shuck plate. See Adjusting Tip Load Switch.
2.2
Adjusting Tip Load Switch
To verify that the tips have been loaded properly is the function of the tip load
switch (brass plunger) located in the front right corner of the pipetting head. As the
disposable tips are loaded, the tip jig makes contact with this plunger pushing it up,
breaking the connection in the switch. This indicates that the tips are loaded
properly to the right height.
This tip load switch needs to be perfectly adjusted for the correct operation. There
is an adjustment screw in the middle of the brass plunger. To adjust this screw, a
9/64” Allen wrench is needed. You must hold the brass plunger stationary while
you turn the adjustment screw. The screw has a spring on it to keep tension on
the screw threads preventing vibration from altering the setting you have made.
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If the distance between the top of the tip and the shuck plate is more than .062,
you need to turn the Allen wrench ½ turn, counter clockwise. If the tips are too
tight to the shuck plate, turn the Allen wrench ½ turn clockwise. In either situation,
do not change this setting more than ½ turn, without retesting the tip load function.
3.0 Quadra Positive Displacement Pipettor Heads
Tomtec offers 4 positive displacement pipettor heads in addition to the air
displacement design. The 96 SV and 384 SV heads offer 60µL capacity. The 96 LV
and 384 LV heads offer 200µL capacity. These positive displacement pipettors use a
fixed stainless steel dispensing needle that is titanium nitrite coated and extends
the full length of the tip bore. This eliminates the dead air volume, common to an
air displacement pipettor design.
Why is this important? On an air displacement pipettor, the dead air volume, at a
minimum, is equal to the tip volume. A 200µL tip volume has 200µL of air between
the orifice and the piston. The 200µL SV positive displacement design, has a 2µL air
gap between the end of the piston and the tip orifice.
Both are governed by the general gas law, which is PV=nRT. With air at the
same temperature, this becomes P1V1=P2V2, where 1 is at the start of an aspirate
and 2 is after the piston has moved. For illustration, assume your finger has blocked
the orifice. Aspirate
200µL with both pistons. For the air displacement pipettor V1 = 200µL the dead
volume of the tip and P1 = 14.7psi atmospheric. V2 becomes 400µL, thus P2
becomes 7.35psi, and which is the negative pressure created to aspirate liquid
through the orifice.
Using the same scenario for the positive displacement design, VI = 2µL and Pi =
14.7. However, V2 now is 202µL. P2 = P1V1/V2 or 0.14psi, which is a much higher
vacuum to aspirate the liquid into the tip. In summary, the air displacement pipettor,
moved the full
tip volume of 200µL to reduce the pressure by half. The positive displacement design
only moves 2µL to reduce the internal pressure by the same amount. This same
logic, applies to the dispense motion. Thus, the positive displacement design, more
equally matches the piston motion to the fluid motion in the tip. Piston motion is what
the software logic controls.
Tomtec has developed an ultrasonic tip washing system for positive displacement
pipettor heads. It is also very effective with the polypropylene disposable tips. A
peristaltic pump provides fresh water, entering the bottom of the wash station and
overflowing the weir at the top. The water in the station is excited with an ultrasonic
transducer, which cleans by cavitation at the tip surface. The tips aspirate fresh water
from the bottom portion of the station, and then dispense it above the liquid level,
allowing the effluent to wash over the top and be carried to the drain.
In all of the testing by Tomtec and others, we have not been able to measure any
carry-over with Fluorescence, Colorimetric, and Scintillation detection. In all cases,
the carry over wells were equivalent to the blank wells. This means if there was
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carry over, it was below the detection limits of the measuring instrument. The last
test was with a Micro Mass Spectrometer. There was no detectable carry-over. The
mass spec detection limits are very low.
On all of Tomtec positive displacement designs, the tip and seal unit is a removable
assembly. It may be removed from the Quadra if more rigorous cleaning is
required. The tip or seals may be replaced as required. This may be accomplished
by the user or the tip and seal assembly may be returned to Tomtec for
reconditioning.
4.0 Quadra 4 DMV Head (Disposable Multiple
Volume)
QUADRA 4 – DMV* PIPETTING HEAD
(*disposable multiple volume)
The Quadra 4 has an optional DMV Pipetting Head that offers the user the ability
to operate the instrument with three different volume pipetting tips with one
pipetting head. The DMV Head uses 250ul, 50ul and 20ul disposable pipetting
tips. The user can select which volume pipetting tips they wish to use by selecting
the color coded tip load fixture in the Quadra 4 Command Software components
menu in upper left corner. The tip load fixture colors are coordinated to the colors
of the pipetting tip bases associated with each volume.
-
Gray – 20ul Pipette Tips – TOMTEC P/N 196-305
Green – 50ul Pipette Tips – TOMTEC P/N 196-307
Blue – 250ul Pipette Tips – TOMTEC P/N 196-306
When the Quadra 4 is programmed to pipette, the user selects proper tip based on
the maximum tip volume capacity for the application being used. This pipette head
is mechanically aligned to this particular Quadra 4 instrument at the factory. This
calibration should not need to be redone during normal operation. Should the
pipetting head or stage be removed or replaced, a new mechanical offset will need
to be determined and installed in the software.
To determine the mechanical offset, the following procedure should be followed.
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TIP LOAD HEIGHT
Tip Load Heights for the 20ul, 50ul and 250ul tips must all be set before the
Electronic Z offset is determined and set in the Quadra 4 Command Software.
1) The DMV pipette head should be installed and aligned in the Quadra 4
instrument.
2) Turn on Quadra 4 POWER
3) Connect and Initialize the Quadra 4
4) Place actual Tip Load Fixture under pipetting head
5) Place proper volume pipette tip rack in Tip Load Fixture
6) Select System
7) Select System Parameters
8) Enter Password 41560
9) Select Board A: XYZ
10) Select Tip Load
11) Select Tip Shuck Height Utility
12) With Proper Pipette Tips on Stacker Nest, Raise stage to Safe Height then
the up arrow until the tips are firmly seated on the Tip Pins without back
driving the stage motor.
13) With tips firmly seated – Select Record Load Height. This will record the
proper stage height
14) Select Safe Height again – this will be the tip shuck (remove) height
15) Select Record Shuck Height
16) Select Set Load Height
17) Select Set Shuck Height
18) In Test Load – Shuck Tips function, select the tip volume for the pipette tips
that have just been loaded
19) Initialize Stage
20) Select OK
21) Return to Tip Utility
22) Select System Utilities
23) Select Shuck Tips
24) Place another set of pipette tips (different volume) on the nest
25) Repeat procedure for other tip volumes
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ELECTRONIC (Z) OFFSET PROCEDURE
1) The DMV pipette head should be installed and aligned in the Quadra 4
instrument.
2) Turn on Quadra 4 POWER
3) Connect and Initialize the Quadra 4
4) Place actual Tip Load Fixture under pipetting head
5) Place proper volume pipette tip rack in Tip Load Fixture
6) Select Utilities
7) Select Pump
8) Select proper volume Tips from drop down menu
9) Select Load Tips
10) Remove pipette tip rack box
11) Close out Utility Window
12) Select System
13) Select set system parameters
14) Enter Password (41560)
15) Select Board A – X,Y,Z - Stage
16) Select Stage z delta alignment utility
17) Select zero Z default
18) Enter Stage Height of 1455
19) Move stage to 1455
20) Evaluate gap between pipette tips and top plate of tip load fixture
21) Jog stage up or down until there is an approximately .005” gap
22) Select set new z delta
23) Select OK
24) Select Initialize
25) Use system utility to test new z delta offset
26) Select OK
27) Select initialize stage
28) Select system utilities
29) Set Stage Height to 1455
30) Move stage to 1455 steps
31) Check for proper stage height (If offset needs additional adjustment - repeat
steps 13 – 23)
32) Select Initialize
33) Replace pipette tip rack box on tip load fixture
34) Shuck Tips
35) Close Utility Window
36) Select OK
37) Select apply changes
38) Select OK
39) Select Reconnect and Initialize Quadra 4
40) Connect and Initialize Quadra 4
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5.0
Setting the Zero Volume Adjustment for Air
Displacement Pipettor Heads
SETTING THE ZERO VOLUME ADJUSTMENT FOR THE
450µL AIR TIP SHUCK PIPETTING HEADS (120932-01 & 120940-01) AND
THE
20µL, 50µL, & 250µL DMV AIR TIP
SHUCK PIPETTING HEADS (120935-01 & 120941-01)
Step
1
Under “System Parameters” select “Pump Zero Volume Alignment Utility”
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026928 – DMV
Zero Volume Setting
Adjustment Gauge
Step 2
Place the supplied adjustment fixture between the Tip Plates as shown in Figure 3
PIPETTING HEAD
FIXTURE #
THICKNESS
450µL AIR TIP SHUCK PIPETTING HEADS
(120932-01 & 120940-01)
026929
0.156”
(3.962
mm)
20µL, 50µL, & 250µL DMV AIR TIP SHUCK PIPETTING
HEADS (120935-01 & 120941-01)
026928
0.110”
(2.794
mm)
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Step 3
In the “calibrate Pump Zero Volume Position” screen Jog down until the head plates just
touch the Fixture.
Step 4
When the head plates are just touching, approx 0.015” (0.381mm clearance), select the
“Record Max Travel” button. The number of steps traveled will be displayed.
Step 5
Select the “Record Max Travel” button. The same number of steps as in Step 4 will be
displayed.
Step 6
Select the “Set Parameters” button.
Step 7
Select the “Initialize Pump” button to re-initialize the pump.
Step 8
Select the “OK” button. The Pump Zero Setting is now set into the factory settings.
6.0 Quadra Shuttle
The concept of the shuttle is what sets the Quadra 4 apart from its competition. The
shuttle can take plates from any of the stackers and move them to and from the
pipettor. On a flat bed pipettor, a separate device, such as a conveyor or robotic
arm, is required. The six stations of the shuttle add to its utility. One station can bring
in the assay plate, while another brings in the compound or test variable plate. One
station may be used for tip washing. This leaves three stations to handle the
reagents for the assay. Using pipeline pipetting complete assays may be completed
in one pass.
The shuttle is driven by two stepper motors, one in the X-direction (left /right) and the
other in the Y-direction (front/ back). Both motors may operate simultaneously
permitting a quick diagonal move when required. The software counts steps
between stations. Stepper motors may loose steps if overloaded. This could
happen if the shuttle is obstructed. This is a nice safety feature but not good as a
control element. The Quadra 4 shuttle’s stopping position is determined by fixed
position sensors at each station. They confirm that the shuttle is where it should be.
A certain level of precision is required for the shuttle to stop on its mark. When the
stage rises the plate nest must fall precisely onto the stage locator pins. Rather than
tune the shuttle position sensors mechanically, they are tuned with software. The
sensor has a wider area of actuation than the final mark. The shuttle calibration
procedure in the manual describes tuning the final stopping mark to its exact point.
The shuttle should stop so that
the stage lifts the plate nest and drops it back cleanly with little to no sideways or
front/back motion. This will center the plate nest on the stage locator pins.
7.0 Stackers
The Quadra 4 stackers are pneumatically actuated by the on board air compressor.
To lift the plate nest from the shuttle into the cassette, air pressure of approximately
20psi is applied to the cylinder. This gives a smooth upward motion. An air pressure
gauge on the inside front panel will indicate these pressure settings. The high and
low air pressure regulators are located there also.
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The escapement that release the plate from the cassette are also pneumatically
driven. The force available with pneumatics provides very reliable escapement
operation. The force of the air cylinder both opens and closes the escapement. As the
stacker carries the plate nest into the cassette, the escapement opens releasing a
plate to the plate nest. As
the plate nest is lowered, the escapement closes to catch the next plate. If all
microplates had the same total height and flange height, then only one setting would
be required. This
not being the case, the timing of the escapement must have some adjustment to
enable handling a wide variety of microplates.
This is accomplished with a sensor mounted on the stacker air cylinder. It tells the
escapement when to open and when to close. It is adjusted with a 3/32 inch Allen
wrench through a clearance hole. The hole is located on the side of each stackers top
plate. Turning the Allen wrench one full turn clockwise moves the sensor down 1/32
inch, two turns equal 1/16 inch. Moving the sensor down will cause the escapement to
stay open longer. Turning the Allen wrench counter-clockwise provides the opposite
effect.
It may be necessary to use trial and error to determine the best position. You should
count turns clockwise or counter-clockwise from your starting point to know where
you are. In general terms, if the plate to be released is still hanging in the cassette, it
probably means the escapement closed too quickly and pinched the plate before it
cleared. In this case, turn the sensor clockwise to cause it to go down, holding the
escapement open longer. If two plates were released, instead of one, then the
opposite is true. Turn the adjusting screw counter-clockwise to cause the sensor to
move up.
The top sensor, mounted on the stacker cylinder PC Board, tells the logic system
that the air cylinder has completed its upward stroke. This initiates the escapement
opening sequence. After a software time delay, the stacker air cylinder up solenoid
valve closes and vents the up motion, allowing the air pressure to drive the stacker
cylinder down. The flag actuates the bottom sensor to indicate the stacker is down
and the system is ready for the next motion. If the bottom sensor is not actuated, the
logic system assumes the stacker is up and will not move the shuttle. An error signal is
generated at that time.
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8.0 Quadra Stage
The stage lifts the plate nests from the Quadra 4 shuttle to the tips. Four pins on
the top of the stage locate into the corresponding holes on the bottom of the nests.
This permits the precision necessary for 384 and 1536 operation. The stage height
is set in the program. It
also has the ability to index in both the X and y direction. This indexing allows
formatting from 96 to 384 or 1536. Since it provides full motion, it also permits serial
dilution testing when used with the 450µL head.
To reformat, the stage must rise above the shuttle to clear it. It also must be below
the tips to clear them. This distance is referred to as the safe height. The stage
must rise to this safe height before it can offset.
The X-motion of the stage is your left and right when you are facing the Quadra 4.
The Y-motion is front to back. The software will automatically set the coordinates for
the required offset for the quadrants of the 384 or the 1536 plate. However, you
may wish to modify
these for a particular plate or application. Use quadrant #1 of the 384 well plate as
an example. The mathematical offset to quadrant # 1 calculates to be 144 X and 144
Y. To
move the plate to the right, causing the tips to be closer to the left side of the well,
enter a negative number for X. To have the plate move forward, causing the tips to
move toward
the back wall of the well, enter a negative number for Y. The numbers you are
entering are actual stepper motor steps. Each step will move the stage 0.0006125
inches.
For Serial Dilution testing, the stage moves approximately half way in both directions
(except Q4 LTD and Q4 SPE). Assume a full plate dilution is to be made in the
twelve tip direction. A single row, of twelve tips, is loaded in Row D. The stage shifts
half way forward so the single row of tips starts at Row A. Then indexes to the rear
so that they end up in Row H. The Quadra 4 stage has a built in safety feature. It is
spring loaded. Should it meet an obstacle with more resistance than the spring force
(approximately 10-15 pounds) the spring collapses and operates a safety switch.
The stage reverses and moves down away from the obstacle. This prevents injury
to you, if your hand is the obstacle, or damage to the tips if a plate is inserted
incorrectly.
When loading a rack of tips on the 450µL head, considerably more force than 1015 pounds is required. This is accomplished with a small magnet embedded in the
tip jig. After the stage has closed the tips on the tip pins, a reed switch is actuated
by the presence of the magnet. This bypasses the safety switch and the stage
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goes to full power loading the tips. The tip switch confirms that the tips are
properly loaded.
The stage reaches its vertical position by counting steps to the number set in the
program. If it becomes jammed or loses steps for any reason, it must initialize. The
stage travels down to its bottom or home position. A sensor is operated at this
point that resets the software counter back to zero.
There is another potential error point. If for some reason, an attempt to lower the
stage is made when it is offset (indexed away from its home position), then it will
not clear the opening in the shuttle. This is recognized by the software and the
stage error message is given. When "escape" is pressed, a "jog stage "window will
appear on the screen. The stage can then be "jogged" (moved) accordingly using
"Page Up" or "Page Down" to remove any obstacle. DO NOT MOVE THE
STAGE BY HAND-DAMAGE MAY OCCUR.
Section V
Suggested Pipetting Methods &
Recommendations Aspirating
The stepper motor driving the pistons has several speeds. Speed 3 is the
fastest and is recommended for aqueous pipetting, with near water like
viscosity. For more viscous liquids, use speed 2 or 1, which is the lowest. At
speed 1, the stepper motor noise increases due to the lower step rate.
If the pistons move too fast, the liquid may not follow that motion, due to the
viscosity. This can create errors in accuracy and precision. If in doubt, watch the
liquid level in the tip, follow the piston motion.
Normally, aspirations are made near the bottom of the container. These values
are set in the Quadra 4 Database. When setting these values, do not pin the tip
to the bottom of the well.
The software provides for "slow withdrawal". The speed with which the tip
leaves the liquid has an effect on drops that can cling to the tip exterior. These
drops are uncontrolled and can affect accuracy. If "slow withdrawal" is checked,
the stage will move down slowly, allowing the surface tension of the reservoir
liquid, to wipe the tip exterior. The stage moves down slowly to the clear height set
in the database, for the plate or reservoir in use. If it moves slowly all of the way,
the database is being used. For aspirating small volumes, accuracy is
improved by pre-wetting the tip. By checking the pre-wet function, the
tip will aspirate and dispense an appropriate volume 3 times, before
aspirating the desired volume.
Polypropylene is basically hydrophobic (repels water), pre-wetting the interior
surface, reduces this hydrophobicity, allowing a more accurate aspirate.
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Experience has shown, repeated use of the tips with ultrasonic washing, reduces
this surface characteristic even more. The caveat is that more liquid will adhere to
the tip e x t e r i o r , t h u s t h e s l o w w i t h d ra w a l f u n c t i o n i s o f v a l u e . If t h e
t i p s e t i s al l o w e d t o dry out, the sequence starts all over. These are small
variations that have been observed. They may or may not have a noticeable
effect on all applications.
1.0 PIPELINE PIPETTING
As described elsewhere, the long narrow portion of the Quadra 450µL tip, may
be used for pipeline pipetting. This allows aspirating several reagents
sequentially, using 10-15µL air gaps, for separation. The separating air gap
must remain within the narrow portion of the tip, which has a capacity
volume of 90µL.
As an example, assume the protocol requires 50µL of buffer, 30 microliters of
reagents A, 20 microliter of reagent B, and 10µL of drug compound. The
Quadra 4 shuttle will present these devices in order to the pipettor head. First
is to aspirate 50µL of buffer, then 10µL of air gap, followed by 30µL of reagent
A, then 10µL of air gap, then 20µL of reagent B, then 10µL air gap and finally
10µL of compound. In addition to speed of throughput, the method can
improve the accuracy of pipetting. The higher volume buffer is used to wash
out the smaller critical volume of drug compound.
2.0 Dispensing
Normally the speed of dispensing is not critical. However, the 3 speeds are
provided. By making the dispense at the top of the well, any separating air
gaps do not create bubbles. These may have an effect on some plate
readers.
After a piston actuated dispense, there may be drops hanging on the tips.
The Quadra 4 software provides several means of returning these drops to
the contents of the well. The simplest and most effective method is a timed
dispense function. This is made at the clear height of the well, as set in the
database. Low air pressure from the internal air tank is discharged through
the tips, clearing the orifice.
Another method is tip touch off. If this is checked, the stage will move the
plate over to cause the tip to touch the sidewall of the well. This distance is
defined in the plate database.
The third option is to dip the tip back into the liquid contents of the well to
allow surface tension to remove the clinging drop. The choice of the
methods is dependent on the application.
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3.0 Mixing
In many applications, it is desired to mix the contents of the well, following a
dispense. The Quadra 4 provides several mixing functions.
4.0 Titration
This is simply making multiple aspirate and dispense moves. It can be
accomplished with any of the Quadra 4 pipettor heads. It is recommended
that the aspirate be made from the bottom of the well, and dispensed at the
top to turn the liquid over. Aspirate approximately 70% of the well contents
volume each time.
5.0 Bubbling
This method is to aspirate the total tip volume of air (at the clear height) then
dispense this air to the bottom of the well using speed 3. This can be a very
effective method of mixing. It may not be suitable for those reagents
containing a detergent or other surfactant due to foaming.
6.0 Injection
This method is restricted to the air displacement pipettor heads that have the
ability to blow pressurized air through the tip orifices.
The desired volume is aspirated in the tips. The tips are moved above the
liquid surface of the well. A timed dispense function is made, applying air
pressure to the tips, ejecting the tip contents. The three parameters
controlling this function are the volume in the tip, the dispense height above
the liquid surface, and the applied air pressure.
These must be controlled to keep below the splashing level. Part of the liquid
volume may be dispensed with piston motion, using air pressure to dispense
the last. The air pressure is set by the black knob on the side that regulates
the air pressure in the internal air source. 1 to 2psi is recommended as a
starting point.
Section VI
Pipetting with the Positive Displacement
Pipettor Head
The 96 SV, 96 LV, 384 SV, and 384 LV pipettor heads are all positive
displacement designs. The 60µL SV heads were designed to aspirate
and dispense small volumes down to 0.5µL. They use a swaged tip
orifice. The 200µL LV pipettors are designed for higher volumes (200µL)
with some sacrifice at the 1µL level.
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The positive displacement designs, do not have the tip blow through
feature of the air displacement design. To clear the tip orifice of
residual, they use piston derived blow out air and surface contact
touch off. The dip tip function after a dispense is also very useful.
The SV heads are capable of aspirating and dispensing 0.5uL volumes
with a CV of 2-3% providing careful pipetting techniques are used.
These are detailed as follows. They are primarily based on transferring
0.5µL of compounds in 100% DMSO into a 25-50µL assay.
1.0 Pipetting 0.5µl with Positive
Displacement
When transferring 0.5µL aliquots the best CV is obtained by using
pipeline pipetting. The other reagents in the assay are aspirated into
the tips, first using air gap separation. Aspirate the 0.5µL aliquot last,
do not separate it with an air gap. Keep it in contact with the last
reagent. This utilizes surface tension between the two liquids to
support the 0.5µL aliquot.
When transferring 0.5µL it is recommended that the tip exterior be
rinsed in the tip washing station. If only 50 nanoliters is on the tips
exterior, it represents a 10% error over the desired 500 nanoliters.
Before dipping the tips, aspirate a 2-3µL air gap at the orifice. This will
prevent the surface tension of the rinse water from attracting and
diluting the 0.5µL aliquot. Tests have shown this method of pipetting
small volumes will result in a CV of 2-3% when using the 96SV or 384
SV tips.
The above applies to only the Teflon coated needles of the 96SV and
384SV head. These results cannot be obtained with the polypropylene
tips of the 450uL head. At 10µL they will consistently provide a CV of
2%. At the 2-5µL level, the CV may be expected to be in the 3-5%
range with careful pipetting techniques.
2.0 Dispensing 0.5µl spots - aqueous
There are some applications that 0.5µL aliquots need to be dispensed
into a dry well without the use of washing out. Testing has shown the
following when pipetting aqueous solutions.
a. Set the tip height carefully to obtain 100% touch off of all
needles. If set too loose or too tight to the plate well bottom, some
spots will be missed. There is about a 0.010 inch (thickness of three
sheets of copy paper) variation that is allowed for 100% touch off.
b. Using 2µL of blow out air will improve the CV of the spot.
c. Dispense the 0.5µL aliquot with the tips in free air above the
wells. Then move the tips down to the touch off height and
dispense the 2µL of blow out air.
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d.
Do not try to rinse the tip exterior as described before. If the 0.5µL
is moved away from the orifice, it will be lost without the supporting liquid
above. If rinsed with the 0.5µL at the orifice the rinse water will capture
it.
3.0
Dispensing 0.5µl spots – DMSO
DMSO (Dimethyl Sulfoxide) is not as hydrophobic to Teflon as water is.
When aspirating DMSO, small droplets may cling to the tip exterior. On
the dispense, if the aliquot touches the tip exterior that is wetted by a
droplet, the small aliquot will follow the wetted surface up the tip
exterior. Corrective action is as follows.
1.
Use the slow withdrawal function in the program when aspirating
DMSO from the source. The slow withdrawal of the tips from the DMSO
allows surface tension to clear the tip exterior of clinging droplets.
2.
Make the dispense when in firm contact with the bottom of the
plate. The tips should press firmly to the well bottom (just short of
stage height error). Do not dispense the drop in mid air as with
aqueous.
3.
Keep the tips clean.
4.0
Protocol Verification
1)
Having completed a pipetting protocol, it is prudent to evaluate it,
with food coloring to represent the various reagents. This will
provide a visual image of the protocol. Look for the following
criteria:
2)
Is the liquid level uniform within the tips at the end of an aspirate
on dispense function?
3)
4)
On a mix function, is liquid level following the piston motion? If
not, slow the aspirate/dispense speeds.
Are the tips being cleared on a dispense?
5)
Is there liquid clinging to the tip exterior?
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Section VII
Bioanalytical Applications
1.0 Aqueous and Organic Pipetting
The features of the Quadra 4 450µL pipettor head has made it the
dominant instrument for BioAnalytical applications. It lives on its history
of use in the Quadra 96 Model 320 and Quadra 3. Quadra 4 platform is
the third generation design and may be supplied with stackers and
automated filtration for full walk away automation. The other option is
the lower cost, non-stacker equipped, Quadra 4 SPE Model.
When pipetting volatile organic fluids, it is necessary to first fill the tip
head space (dead air volume) with the organic vapors. Otherwise, as
the organic evaporates to equilibrate the head space, this decreases
the negative pressure (tip vacuum) that is holding the column of liquid
in the tip. The result is the tips drip.
To prevent the dripping action, first use the mix function to aspirate and
dispense 450µL one or more times, depending on the organic involved.
A more volatile organic such as MBTE may require 2 or 3 mix cycles.
Actonitrile (ACN) may only require 1 or 2. An expedient for less volatile
liquids, such as the alcohols, is to aspirate the desired volume followed
with a 10µL air gap. The organic may not have displaced the air gap
before the final dispense is made.
An even more important consideration is to clear the tips of the organic
vapors prior to pipetting aqueous liquids. This is easily accomplished with the
Quadra 4 by making 2 or more time dispense functions. This opens the pinch
clamp, allowing air pressure, from the internal air tank, to clear the tips of
organic vapor. If the organic vapors are not cleared, the well condense in the
head space which has the opposite effect. The condensing vapors decrease
the negative pressure in the tip head space, aspirating a higher volume than
required. This affects the accuracy of the pipetting.
2.0 Protein Precipitation, Liquid/Liquid, And SPE
For some applications the required mixing of protein and organic may be
accomplished on the Quadra 4 eliminating the secondary operation of
vortexing. Using a deep well plate, (1mL or 2mL), mixing may be
accomplished using bubbling or injection as described before. Before
aspirating free air for bubbling, make a time dispense to clear the orifices of
precipitated protein. The use of protein filter plates (i.e. Waters Sirocco,
Varian, Captiva, etc.) can eliminate the centrifuge requirement for separation.
Tomtec's automated vacuum box accessory can provide full walk-away
automation for filtration protocols.
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3.0 Protocol Or Program Security
The flexibility in programming the Quadra 4 has many advantages. The ease of
alterations can affect the performance of specific programs. The solution for
this problem is provided by the program checksum. The program checksum is
a 32 bit number that adds all of the digital steps in a program. It is recalculated
each time that program is called for operation. If the checksum matches the
original validated program, the Quadra 4 will execute exactly the same
program. The accuracy is one bit in 232 or 4 trillion sum.
A checksum for every program is calculated at the time the program is called.
The checksum is composed of two separate additions - the program itself and
the Quadra 4 parameters. If the checksum matches the check sum originally
recorded for a validated program, it assures the operator the Quadra 4 will run
the exact same protocol.
If there is a mismatch, the checksum can be shown in its two parts - program
and parameters. If the program checksum has a mismatch, the original
program may be reloaded. This assumes an original validated copy is retained.
If the mismatch is in the instrument parameters, then it is necessary to check
each one for a mismatch since they are individually set.
While the Quadra 4 calculates the checksum each time any program is called,
it does not verify that it matches a previous record. This must be accomplished
external to the Quadra 4 normal software configuration.
4.0 Calibration For The Quadra 4
On the Quadra pipettor heads, the stepper motor moves the pistons a precise
distance with each step. Calibration is simply telling the stepper motor how
many steps are required for each volume to be displace All of the Quadra
pipettor heads use a similar method for calibration. The 450µL head is used as
an example.
The first step is to determine how many steps are required to deliver a
measured volume. In the "factory settings”, the steps per microliter factor is set
to 16 steps per microliter. The "backlash" factor must be set to zero. With the
display set to transfer 10µL, the software will move the pistons 160 steps. Both
to aspirate and dispense. The volume dispensed is accurately weighed.
Assume water is the liquid and the weight was 0.0089 grams or 8.9µL. Thus,
160 steps gave 8.9µL or 17.977 steps, per microliter. This becomes the calibration factor for 10µL.
An air displacement pipettor covering a wide volume range, such as the
Quadra 4 (10µL to 450µL), does not have a linear calibration curve. The dead
air volume in the pipettor, at a minimum, is equivalent to the tip volume
(450µL). The physical laws of nature govern the compressibility of this air
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column and its displacement of liquid. The higher volumes from 100µL to 450µL
are nearly linear. The physical factors have a greater impact on the smaller
volume in the range. Thus, the accuracy of calibration is a function of how many
points on the curve are measured and how many are interpolated. This is
determined empirically.
Another point of consideration is, how many weighing's of each value is
required for the desired accuracy. This is a reflection of the repeatability of the
pipetting system. Tomtec's testing has been to measure 5 values at each point.
From 5µL to 450µL, the CV of 5 values has consistently been better than 0.5%.
This implies fewer reading could be used provided they are consistent without
"flyers". The "flyers" are outlayers that are due to other factors i.e., faulty
weighing, non-wetting, etc.
On the 450µL backfill head, Tomtec has included a generic calibration curve.
Generic meaning it is an average calibration curve, applied to all 450µL backfill
heads. This generic curve will provide accuracy within 2%, for volumes from
450µL, down to 100µL. From below 100µL to 25µL, the accuracy will be within 3%.
From 25µL to10µL, the accuracy will be within 5%.
The user may create a specific calibration curve for the pipettor head in the
Quadra 4. This will improve the accuracy. With specific calibration, accuracy of
less than 1% can be obtained from 100µL to 450µL. Improvements can also be
made in the lower volume ranges.
To achieve pipetting accuracy within 2%, for volumes from 100µL to 450µL,
Tomtec recommends taking 3 readings at the following volumes: 100µL,150µL,
200µL, 250µL, 300µL, and 450µL. For accuracy within 3%, for volumes from
25µL to 100µL, readings at the following volumes are recommended: 10µL,
15µL, 20µL, 25µL, 30µL, 40µL, 50µL, 70µL, 80µL, 90µL,100µL.
The mean value of the data at each calibration point is entered into the pipettor
calibration table. Tomtec has included an Excel program for calculating mean
and CV values, for the collected data. After entering the calibration data, the
various points should be rechecked for verification. 2 to 3 values at selected
points are recommended.
Since the Quadra pipettor head is calibrated empirically, it may be calibrated for
any pipettable liquid. A specific name is assigned to that calibration curve,
allowing it to be called on demand.
The distinct advantage of Tomtec's 450µL pipettor head is its uniquely wide
volume range from 450µL down to 10µL with disposable tips. This, combined
with our tip blow through feature, has made it the standard for BioAanalytical
applications. This wide range for an air displacement pipettor presents specific
physical requirements that
must be overcome. The calibration curve over this volume range is not a linear
relationship, due to the compressibility of the air column within the disposable
tip.
Stepper motors are used for pipettor drives due to their precision and reliability.
Within its range a stepper motor will move the pipettor piston a precise distance
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for each software pulse it receives. Calibration simply means equating how
many software steps are required to dispense a specific volume.
Due to the compressibility of air, the number of steps to pipette a microliter of
volume is higher at the low volumes that it is at higher volumes. To aspirate or
dispense 10µL may require 18 to 19 steps per microliter. At 450µL, it requires 15
to 16 steps per microliter. If the Quadra 4 is asked to aspirate a volume and
then dispense the same volume, i.e. empty the tip, the calibration curve
handles this well.
Each Quadra 4 instrument will have certain mechanical dimensions. To enable
the use of the common software dimensions, they must be related to the
specific mechanical dimensions, the stage must move on each instrument. This
is accomplished with the "Z" offset factor.
The caveat is if the Quadra 4 is used for aliquoting different volumes from one
aspirate. The calibration curve does not handle this situation as well. Assume
the Quadra 4 aspirated 100µL and then dispensed it in 10µL aliquots. For the
100µL, the calibration curve dictated 16.532 steps, per microliter. The stepper
motor moved the piston 1653 steps, which aspirated 100.3µL. To dispense
10uL, the calibration curve shows 18.678 steps, per microliter or 187 steps for
each 100µL aliquot. After 8 dispenses, the stepper motor had moved 1496,
leaving only 157 steps left, which is insufficient for the last two aliquots.
This simply confirms the standard practice when aliquoting. The last 1 or 2
volume should be dispensed back to the source. Otherwise, cumulative error
will appear in the last volume. The Quadra 4 calibration software, recognizes
this condition, by comparing the asked for aspirated volume to the dispensed
volume to prevent an out of range error from occurring.
In summary, the calibration curve is most efficient at improving accuracy of
pipetting, when aspirating a single volume from the source and dispensing that
volume to the destination. The Quadra's ability to make a timed dispense,
which
blows pressurized air through the tip after the dispense, negates the necessity
of an initial air gap to serve that purpose. The timed dispense is far more
effective at clearing the tip of capillary residual than the initial blow out air gap.
It simply has a higher velocity. The timed dispense function also enables noncontact dispensing.
The example of 10 dispenses from 1 aspirate, is the extreme condition chosen
to demonstrate a point. The Quadra 4 can be used for aliquoting. The user
simply needs to be aware of potential loss of accuracy, when various pipetting
methods are used.
The software database defines the distance from the top of the stacker nest,
resting on the shuttle, to the tips as 1455 steps. Using the utility function, the
stage is indexed up to the point the tips just touch the top of the bare stacker
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nest. Assume this reading is 1465 steps. The "Z" offset (under factory settings)
is then reset from its existing value to -20 steps or less. When retried, the stage
height should now read 1455. This change in the "Z" offset is used to align the
mechanical "Zero" of the Quadra 4 with the "Zero" of the command control
software.
Making a new entry in the database is intuitive. Open to the Quadra 4 and
software initialize the instrument. Click on the database on the tool bar, follow
the description. Prior to entering the plate dimensional data, you must specify
the nest that will be used to carry the plate or reservoir on the shuttle.
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Section VIII
QUADRA 4 ACCESSORIES
Tomtec provides a wide variety of accessories for the Quadra 4 to expand its
value for specific applications.
1.0
Custom Reservoirs
Tomtec will specially fabricate custom reservoirs to match your pipetting
pattern. For example, many clients leave the first row of wells open without
compounds to facilitate adding standards and controls to the pattern. Tomtec
can provide a special reservoir with specific wells joined in the first row.
Using pipeline pipetting, the pipettor would aspirate the standards and
controls from the first row and air in the balance of the tips. When moving to
the compound plate the pipettor will aspirate air in the empty first row and
compounds from the rest.
Keeping Participate Matter in Suspension
There are a number of applications that require dispensing liquids containing
particular matter that must be kept in uniform suspension. If the suspension
is not uniform, then the end results may be affected. Examples are magnetic
beads, SPA beads, cells, etc. Tomtec offers the following, depending on the
applications requirements.
Stirring Reservoir
A self-contained battery operated stirring reservoir nest will sit on the shuttle. It has
three magnetic fleas to stir the contents of the reservoir.
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Constant Level Reservoir
A peristaltic pump is used to circulate fluid to a weir type
reservoir. It maintains a constant liquid level. This
provides a means of controlling the depth of the pipette
tip insertion when aspirating. It eliminates refilling the
reservoir for long runs of plates. It can also be used as
a means of keeping particulate matter re-suspended.
Shaker Nest
This nest has a small DC motor that swings an off center
weight. This imparts a vibrating motion to the microplate
or reservoir residing on the nest. The speed of the motor
and thus the vibrating action is adjustable. This shaker
nest can access the stackers for infeeding or outfeeding.
Live Bottom Reservoir
This is a more sophisticated constant level reservoir for
more exacting applications. The reservoir is fabricated
from Teflon. 24 small holes spaced across the bottom are
the inlet. This keeps the entire bottom area alive and
moving. The overview is a weir that returns to the recirculating source.
Thermal
196-5O6
Retention
Reservoir
Part#
This is simply a block of aluminum that provides thermal
mass for a reservoir. The aluminum may be heated or
cooled before hand and will exchange heat over a
limited period of time. This is not stacker accessible.
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Peltier Nest for heating and
cooling Part# 196-503
This is designed to either heat or cool a flat bottom Teflon
coated aluminum reservoir (340-56). It requires a PID
temperature controller (340-20).
Vacuum Box Part# 196-503
This is a 3 part vacuum box for SPE applications. It provides
means of capturing the diluent in a microplate or deepwell
plate. It is manually operated.
Filtration System for the Quadra 4
Part# 330-34
This consists of the manually operated vacuum box, and a
vacuum regulated mini trap with the Quadra 4 SPE.
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Air Regulator Vacuum Trap – Part#
330-65
This consists of a Quadra 4 Software delivers Vacuum control
regulating the mini-trap.
Fully Automated Filtration
System Part# 330-55
This must be factory installed. This system consists
of a robotic hand for plate transfer. A fully
pneumatically operated vacuum box and a vacuum
regulated mini trap. It is designed to provide a
walk-away automation for all filter plate protocols.
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Section IX
PIPETTOR TIPS
1.0
Tip# 1
When dispensing into the assay plate, set the tip height slightly above what will be
the final volume. This will prevent creating air bubbles in the well due to the air
gaps. Air bubbles may affect the readout with some plate readers. They are difficult
to remove in a 384 well plate.
2.0
Tip #2
The Teflon® coated small volume tips are hydrophobic. Dispense the blow out air
above the final well volume. A small air bubble or droplet may be formed on the tip.
Dip the tips back into the well to wick off this droplet. Polypropylene tips are not as
hydrophobic as Teflon®. When using polypropylene tips, set the tip height for the
blow out air at the top of the well. The Quadra stage also permits touching the
polypropylene tips to the well sidewall. This will not be effective with the stiff Teflon
coated tips. Not only will they not bend but the taper at the orifice prevents it from
touching the side wall.
3.0
Tip #3
When transferring 0.5µL aliquots the best Cv is obtained by using pipeline pipetting.
The other reagents in the assay are aspirated into the tips, first using air gap
separation. Aspirate the 0.5µL aliquot last, do not separate it with an air gap. Keep it
in contact with the last reagent. This utilizes surface tension between the two liquids
to support the 0.5uL aliquot.
When transferring 0.5µL it is recommended that the tip exterior be rinsed in the tip
washing station. If only 50 nanoliters is on the tip exterior, it represents a 10% error
over the desired 500 nanoliters. Prior to dipping the tips, aspirate a 2-3µL air gap at
the orifice. This will prevent the surface tension of the rinse water from attracting and
diluting the 0.5µL aliquot. Tests have shown this method of pipetting small volumes
will result in a Cv of 2-3% when using the 96SV or 384 tips.
The above applies only to the Teflon® coated needles of the 96SV and 384 head.
These results cannot be obtained with the polypropylene tips of the 450µL head. At
10µL they will consistently provide a Cv of 2%. At the 2-5µL level, the Cv may be
expected to be in the 3-5% range with careful pipetting techniques.
4.0
Tip #4
There are some applications that 0.5µL aliquots need to be dispensed into a dry well
without the use of washing out. Testing has shown the following:
a. Set the tip height carefully to obtain 100% touch off of all needles. If set too loose
or too tight to the late well bottom, some spots will be missed. There is about a
0.010 inch (thickness of three sheets of copy paper) variation that is allowed for
100% touch off.
b. Using 2µL of blow out air will improve the Cv of the spot.
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c. Dispense the 0.5µL aliquot with the tips in free air above the wells. Then move
the tips down to the touch off height and dispense the 2µL of blow out air.
d. Do not try to rinse the tip exterior as described in Tip # 3. If the 0.5µL is moved
away from the orifice, it will be lost without the supporting liquid above. If rinsed
with the 0.5µL at the orifice the rinse water will capture it.
5.0
Tip #5
When mixing reagents in the well, aspirate from the bottom of the well and dispense
above the final liquid level. This will turn the liquid over. It is recommended that you
mix with 70-75% of the well volume.
5.1 Auxiliary Connector Panel
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5.2 Serial In- For use with the optional Barcode Reader Installation kit
(part # 330-10). Connect the jumper cable to "Barcode Out".
5.3 Barcode Logging Functions- Are found under file/logging
options in the toolbar.
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Section X
CALIBRATION OF THE QUADRA 4® 450µL 96 POSITION
PIPETTING HEAD
Tomtec supplies all Quadra 4® models with a calibration curve based on pipetting
water. Other fluids can be used to create additional calibration curves by the user.
On the 450uL disposable tip head, this curve provides a pipetting accuracy error of
less than 2% for volumes from 450uL down to 100uL. From 99uL to 50uL, the
accuracy error is less than 3%. From 49uL down to 10uL, the accuracy is be 5%
or less. This accuracy is based on the mean volume delivered by all 96 tips. This
is determined by accurately weighing the total volume dispensed and dividing by
96 to achieve the mean volume. Each individual tip may have a wider accuracy
tolerance than the mean value.
The calibration curve is most efficient at improving accuracy of pipetting, when
aspirating a single volume from the source and dispensing that volume to the
destination. The Quadra's ability to make a timed dispense, which blows
pressurized air through the tip after the dispense, negates the necessity of an
initial air gap to serve that purpose. The timed dispense is far more effective at
clearing the tip of capillary residual than the initial blow out air gap. It simply has a
higher velocity. The timed dispense function also enables non-contact dispensing.
The 450uL pipettor head has wide volume range from 450uL down to 10uL with
disposable tips. This wide range for an air displacement pipettor presents specific
physical requirements that must be overcome. The calibration curve over this
volume range is not a linear relationship, due to the compressibility of the air
column within the disposable tip.
Due to the compressibility of air, the number of steps to pipette a microliter of
volume, is higher at the low volumes that it is at higher volumes. To aspirate or
dispense 10uL, may require 18 to 19 steps per microliter. At 450uL, it requires 15
to 16 steps per microliter. If the Quadra 4® is asked to aspirate a volume and then
dispense the same volume, i.e. empty the tip, the calibration curve handles this
well.
The caveat is if the Quadra 4® is used for aliquoting different volumes from one
aspirate. The calibration curve does not handle this situation as well. Assume the
Quadra 4® aspirated 100uL and then dispensed it in 10uL aliquots. For the 100uL,
the calibration curve dictated 16.532 steps per microliter. The stepper motor
moved the piston 1653 steps, which aspirated 100.3uL. To dispense 10uL, the
calibration curve shows 18.678 steps per microliter or 187 steps for each 10uL
aliquot. After 8 dispenses, the stepper motor had moved 1496, leaving only 157
steps left, which is insufficient for the last two aliquots.
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This simply confirms the standard practice when aliquoting. The last 1 or 2
volumes should be dispensed back to the source. Otherwise, cumulative error will
appear in the last volume. The QUADRA 4 calibration software, recognizes this
condition, by comparing the asked for aspirated volume to the dispensed volume
to prevent an out of range error from occurring.
A calibration curve for the Quadra 4® may be created for any pipetttable liquid.
The procedure is the same. Aspirate and dispense the desired volume with a
setting of 16 steps per microliter. Gravimetrically measure the actual volume
dispensed. Divide the actual volume by the number of steps used to define the
calibration point. The calibration curve is not linear on an air displacement
pipettor. Therefore it is necessary to define a sufficient number of points on the
curve to achieve the desired accuracy, over the desired pipetting range of
volumes.
The data is entered into the software with a fluid class name i.e. water, methanol,
acetonitrile, DMSO, etc. The fluid class name is not case sensitive. It may be
entered in upper or lower case letters. The date of the calibration is also entered.
The Quadra 4® programs are created using the fluid class name i.e. water. Any
time in the future this program is called, it will use that fluid class calibration. The
actual data will be the default value of the last data entry for that fluid class i.e.
water (6-16-06). Other fluid classes, or other dated curves, may be selected at the
time the program is executed. The default value will be the last value entered for
that fluid class, unless another selection is made at the time of running the
program.
1.0
EQUIPMENT REQUIRED
1. Four place Calibrated Precision Balance.
2. ISO Grade 3 distilled or de-ionized water.
3. Weighing Vessels (Qty 78 – 48 for creating a calibration curve and 30 for
verifying the calibration curve).
4. Cat. No. 320-50 Tip Loading Jig (supplied with the QUADRA 4 series).
5. Cat. No. 196-130-00 Polypropylene Reagent Reservoir with 3 baffles (supplied
with the QUADRA 4 series).
6. Cat. No. 196-205 QUADRA Pipetting Tips (supplied with the QUADRA 4
series).
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2.0 WEIGHING VESSELS
Weighing Vessels may be purchased commercially or user made from light weight
aluminum foil. The lid of a Microtiter Plate or a 96 well Flat Bottom Microtiter Plate
may also be used.
For the “CALIBRATION CURVE” procedure, label each Weighing Vessel with the sample
number consisting of the volume to be Calibrationed and the pipetting number (1, 2, 3) i.e.,
5-1, 5-2, 5-3, … for each of the 16 SET VOLUMES listed below required for creating the
calibration curve.
THIRD PIPETTING
5-1
SECOND
PIPETTING
5-2
10µL
10-1
10-2
10-3
15µL
15-1
15-2
15-3
20µL
20-1
20-2
20-3
30µL
30-1
30-2
30-3
40µL
40-1
40-2
40-3
50µL
50-1
50-2
50-3
60µL
60-1
60-2
60-3
70µL
70-1
70-2
70-3
90µL
90-1
90-2
90-3
100µL
100-1
100-2
100-3
125µL
125-1
125-2
125-3
200µL
200-1
200-2
200-3
300µL
300-1
300-2
300-3
SET VOULME
FIRST PIPETTING
5µL
5-3
450µL
450-1
450-2
450-3
Using the 4 decimal place balance, record the tare weight of each weighing Vessel.
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For the “VERIFICATION OF THE CALIBRATION CURVE” procedure, label each
Weighing Vessel with the sample number consisting of the volume to be Calibrationed and
the pipetting number (1, 2, 3) i.e., 10-1, 10-2, 10-3, … for each of the 10 SET VOLUMES
listed below required for the verification of the calibration curve.
SET VOULME
FIRST PIPETTING
SECOND
PIPETTING
THIRD PIPETTING
10µL
10-1
10-2
10-3
25µL
25-1
25-2
25-3
50µL
50-1
50-2
50-3
75µL
75-1
75-2
75-3
100µL
100-1
100-2
100-3
125µL
125-1
125-2
125-3
150µL
150-1
150-2
150-3
250µL
250-1
250-2
250-3
350µL
350-1
350-2
350-3
400µL
400-1
400-2
400-3
Using the 4 decimal place balance, record the tare weight of each weighing Vessel.
3.0 EVAPORATION
For small volumes, below 50µL, errors due to evaporation of the Calibration Liquid during weighing
should be taken into consideration. To keep the error due to evaporation as small as possible the
following should be considered:
A balance with the appropriate accessories such as an evaporation trap.
Pipetting to weighing Cycle Time.
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4.0 PIPETTING TO WEIGHING CYCLE TIME
The weighing Cycle Time (time required to complete the weighing of one dispensed
volume) should be kept to a minimum. It should not exceed 60 seconds. It is also important
that it is regular, both within each cycle and as far as possible from cycle to cycle.
5.0 SETUP PROCEDURE
1. Power up the QUADRA 4. After startup the “QUADRA 4 COMMAND SYSTEM”
program should be displayed. If not, select and open the “QUADRA 4 COMMAND
SYSTEM” program from the desktop icon.
2. Click on the “CONNECT TO SYSTEM” button.
3. Click on the “INITIALIZE” button to initialize the QUADRA 4 .
4. Verify the Air Pressure for BLOW-OUT is set between 1.00 and 1.25psi.
5. Verify that the Backlash Factor, per factory specifications has been set into the
Instrument Parameters of the Quadra. (The factory set Backlash Factor number can
be found on the pipetting head and/or the Factory Setting sheet located in the pocket
on the inside of the front cover).
6. Verify that the µL Factor (16) has been set into the Instrument Parameters of the
Quadra 4.
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Tomtec
6.0 ADDING THE NEW “CALIBRATION CURVE” INTO THE QUADRA COMMAND
SYSTEM SOFTWARE FROM DATA POINTS
Open the “Step 3 Calibration Summary” spreadsheet
1.
Open the “QUADRA 4 COMMAND SYSTEM” program installed on the Computer.
2.
3.
4.
In the upper menu bar select “UTILITIES”.
On the pull down menu under “UTILITIES” select “PUMP CALIBRATION”.
5.
On the “PUMP CALIBRATION” window select “NEW”, a popup window will
ask for LIQUID TYPE, type in “WATER (XX-XX-XX)” where the XX-XX-XX is
today’s date. Select the “OK” button.
6.
A popup window “ENTER NEW PUMP CALIBRATION TABLE DATA” will
appear. Fill in the values for “EXPECTED VOLUME” and “ACTUAL
VOLUME” from “Step 3 Calibration Summary” spreadsheet. Select the
“ADD” button.
7.
Continue filling in the “EXPECTED VOLUME” and “ACTUAL VOLUME”
boxes for all 16 SET VOLUME results from “Step 3 Calibration Summary”
spreadsheet. Select the “OK” button.
8.
Checking the “SHOW REFERENCE CURVE” box will graphically display the
new calibration curve.
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Tomtec
7.0 “CALIBRATION TEMPLATE – Q4” SPREADSHEET
1. Open the “CALIBRATION TEMPLATE – Q4” Spreadsheet, making sure the “ENABLE
MACROS” button is selected.
2. On the “Step 1 Define µL Volume to Calibration” spreadsheet select the RED
“Populate Calibration Vol. Example Data” button.
3. Save the spreadsheet by selecting “SAVE AS” and naming the file by the LIQUID
CLASS, and date i.e “WATER (XX-XX-XX)” where the XX-XX-XX is the today’s date.
4. Open the “Step 2 Calibration Data” and at the top of the spreadsheet fill in the
following information:
TODAY’S DATE
TIME
TEMP º C (To nearest 0.2ºC)
HUMIDITY % (To nearest 10%)
BAROMETRIC PRESSURE in. Hg
HEAD SERIAL NUMBER
5. Fill in the tare weight of each Calibration weighing Vessel in the area marked “TARE
WT.”.
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Tomtec
8.0 CREATING A NEW “CALIBRATION CURVE” WITH WATER
1. The “CALIBRATION CURVE” procedure consists of pipetting a set volume of the
Calibration Liquid and creating a correction curve to increase Pipetting Head Accuracy.
The procedure uses 16 pipetted set volumes pipetted 3 times.
For each set volume, the Expected Weight is compared to the Dispensed Weight as a %
ERROR using the following formula:
(EXPECTED WEIGHT / PER WELL) – (DISPENSED WEIGHT / 96)
------------------------------------------------------------------------------------ X 100 = % ERROR
(EXPECTED WEIGHT / PER WELL)
NOTES:
1. 1 GRAM = 1mL
2. Tips are pre-wetted prior to performing the following Calibrations.
3. The time between dispensing and weighing should be minimized to avoid
errors due to evaporation.
4. The average % Error of the 3 weights for each set volume is used in creating
the calibration curve data points.
5. Tomtec provides a spreadsheet to calculate the % ERROR. It also calculates
the mean, standard deviation and CV of the specific samples that are entered
for each volume.
2. PRE-WET TIPS
1. Place the Tip Loading Jig (Cat. No. 320-50) with a rack of QUADRA Pipetting Tips
(Cat. No.196-205) on position 1 of the shuttle
2. Load the rack of QUADRA Pipetting Tips using the “LOAD TIPS” Program.
3. Place a Plate / Reservoir Nest (Cat. No. 320-30) with a Polypropylene Reagent
Reservoir (Cat. No. 196-130-00) containing the Calibration Liquid onto the shuttle,
Position 6.
4. Place a Plate / Reservoir Nest (Cat. No. 320-30) onto the Shuttle, Position 5.
5. Run the “PRE WET” program.
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Tomtec
3. CREATING CALIBRATION CURVE DATA POINTS
1. Place the tared Weighing Vessel labeled 5-1 onto the Plate / Reservoir Nest at
Position 5 of the Shuttle.
2. Run the “CAL 5µ
µL” program to dispense 5.0µL of the Calibration Liquid into the
tared Weighing Vessel labeled 5-1
3. After the Calibration Liquid is Dispensed, immediately obtain the gross weight of
the sample and Weighing Vessel labeled 5-1. Record the weight onto the supplied
spreadsheet.
spreadsheet.
spreadsheet.
10. Continue this routine using the supplied programs for the remaining 15 SET
VOLUMES (10µL to 450µL ) doing 3 pipettings of each volume.
11. Shuck and dispose of the QUADRA Pipetting Tips using the “SHUCK TIPS”
program.
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Tomtec
9.0 CREATING A “VERIFICATION CURVE”
1. The “VERIFICATION OF THE CALIBRATION CURVE” procedure consists of pipetting
a set volume of the Calibration Liquid and crating a correction curve to increase Pipetting
Head Accuracy. The procedure uses 10 pipetted set volumes pipetted 3 times
For each set volume, the Expected Weight is compared to the Dispensed Weight as a %
ERROR using the following formula:
NOTES:
1. 1 GRAM = 1µµL
2. Tips are pre-wetted prior to performing the following Calibrations.
3. The time between dispensing and weighing should be minimized to avoid
errors due to evaporation.
4. The average % Error of the 3 weights for each set volume is used in creating
the Verification Curve data points.
5. Tomtec provides a spreadsheet to calculate the % ERROR. It also calculates
the mean, standard deviation and CV of the specific samples that are entered
for each volume.
2.
PRE-WET TIPS
1. Place the Tip Loading Jig (Cat. No. 320-50) with a rack of QUADRA Pipetting Tips
(Cat. No.196-205) on position 1 of the shuttle
2. Load the rack of QUADRA Pipetting Tips using the “LOAD TIPS” program.
3. Place a Plate / Reservoir Nest (Cat. No. 320-30) with a Polypropylene Reagent
Reservoir (Cat. No. 196-130-00) containing the Calibration Liquid onto the shuttle,
Position 1.
4. Place a Plate / Reservoir Nest (Cat. No. 320-30) onto the Shuttle, Position 6.
5. Run the “WET TIPS” program.
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Tomtec
3. CREATING “VERIFICATION OF THE CALIBRATION CURVE” DATA POINTS
1.
Place the tared Weighing Vessel labeled 10-1 onto the Plate / Reservoir Nest at
2.
Run the “VER 10µ
µL” program to dispense10.0µL of the Calibration Liquid into the
3.
After the Calibration Liquid is Dispensed, immediately obtain the gross weight of
the sample and Weighing Vessel labeled 10-1. Record the weight onto the
supplied spreadsheet.
4.
5.
Run the “VER 10µ
µL” program to dispense 10.0µL of the Calibration Liquid into
the tared Weighing Vessel labeled 10-2
6.
7.
8.
Run the “VER 10µ
µL” program to dispense 10.0µL of the Calibration Liquid into
the tared Weighing Vessel labeled 10-3
9.
10.
Continue this routine using the supplied programs for the remaining 9 SET
VOLUMES (25µL to 350µL ) doing 3 pipettings of each volume.
11.
Shuck and dispose of the QUADRA Pipetting Tips using the “SHUCK TIPS”
program.
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Tomtec
4. The “AVG % ERROR” on the spreadsheet for the SET VOLUMES run in the
“VERIFICATION OF THE CALIBRATION CURVE” procedure MUST fall within the
AVERAGE % ERROR range listed in the chart below:
AVERAGE %
SET VOLUME
ERROR
≤ 5.0%
10.0µL to 49.9µL
< 3.0%
50.0µL to 99.9µL
< 2.0%
100.0µL to 450.0µL
5. If any SET VOLUME fails the VERIFICATION, redo the “CALIBRATION CURVE”
procedure and repeat the “VERIFICATION OF THE CALIBRATION CURVE” procedure.
10.0 “CALIBRATION TEMPLATE – Q4” FOR THE VERIFICATION CURVE
1. Open the “Step 4 Verification Data” and at the top of the spreadsheet fill in the
following information:
TODAY’S DATE
TIME
TEMP º C (To nearest 0.2ºC)
HUMIDITY % (To nearest 10%)
BAROMETRIC PRESSURE in. Hg
HEAD SERIAL NUMBER
2. Fill in the tare weight of each weighing Verification Vessel in the area marked “TARE
WT.”.
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Tomtec
11.0 CALIBRATION INSTRUCTIONS FOR THE QUADRA 4 SERIES 450µL PIPETTING
HEAD
11.1 DEFINITIONS
CALIBRATION
Is defined as the process by which the correlation between instrument
response and actual value of a measured parameter is determined.
CALIBRATION
CURVE
Is defined as the curve which plots the amount of liquid dispensed versus
the expected amount. Also known as a Standard Curve.
INACCURACY
Is defined as the difference between the DISPENSED volume and the
NOMINAL or expected volume to be dispensed. (ALSO CALLED
“SYSTEMATIC ERROR”)
11.2 LIQUID USED FOR CALIBRATION CURVE SET VOLUMES
1. The Calibration Liquid for a “Fluid Class of Water” should be distilled or de-ionized water
conforming to grade 3 as specified in ISO 3696, degassed or air-equilibrated. The water
shall be at room temperature (20º C).
12.0 AMBIENT CALIBRATION CONDITIONS
The following Ambient Calibration Conditions should be complied with:
AIR
TEMPERATURE:
25 °C to 35 °C (constant ± 0.5 °C)
RELATIVE
HUMIDITY:
50 % to 65 %
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Tomtec
12.1 IMPORTING AN EXISTING “CALIBRATION CURVE”
Open the “QUADRA 4 COMMAND SYSTEM” program installed on the Computer.
In the upper menu bar select “UTILITIES”.
On the pull down menu under “UTILITIES” select “IMPORT DATA”.
Select the USB drive and select “WATER (XX-XX-XX).csv”.
A popup window will ask for LIQUID TYPE, type in “WATER”. Select the “OK”
button.
6.
Checking the “SHOW REFERENCE CURVE” box will graphically display the new
calibration curve.
175
Tomtec
Section XI
SOME PROPERTIES OF LIQUIDS THAT AFFECT PIPETTING
ACCURACY
1.0 THE RELATIONSHIP BETWEEN LIQUID AND GASEOUS STATES OF A SOLVENT
HAVE AN EFFECT ON PIPETTING
1.
VOLATILITY:
The property of changing readily from a solid or liquid to a vapor
2.
VAPOR
PRESSURE:
The pressure exerted by a vapor; often understood to mean
saturated vapor pressure
3.
PARTIAL
PRESSURE:
The pressure that one component of a mixture of gases would exert
if it were alone in a container.
2.0 SOME OTHER PROPERTIES OF LIQUIDS THAT HAVE AN EFFECT ON PIPETTING
1.
DENSITY:
The density of a liquid will significantly impact the ability of an air
displacement pipetting mechanism to aspirate the liquid into the
pipette tip
2.
COMPRESSION:
Different liquids and gases have different compression coefficients
that can effect pipetting, especially when the flow is partially
obstructed
3.
VISCOSITY:
This contributes to the resistance-to-flow of a liquid
4.
SURFACE
TENSION:
Is an effect within the surface layer of a liquid that causes the layer
tobehave as an elastic sheet
5.
WETTING:
Refers to the contact between a fluid and a surface, when the two
are brought into contact. When a liquid has a high surface tension
(strong internal bonds), it will form a droplet, whereas a liquid with
low surface tension will spread out over a greater area (bonding to
the surface). On the other hand, if a surface has a high surface
energy (or surface tension), a drop will spread, or wet, the surface. If
the surface has a low surface energy, a droplet will form. This
phenomena is a result of the minimization of interfacial energy. If the
surface is high energy, it will want to be covered with a liquid
because this interface will lower its energy, and so on.
NOTE: All of the above properties are affected by temperature
176
Tomtec
3.0 TO COMPENSATE FOR THESE PROPERTIES OF LIQUIDS, THE ASPIRATE
AND DISPENSE STEPS HAVE OPTIONS TO IMPROVE PERFORMANCE
1.
Pre-filling the head space with saturated vapors of a volatile solvent. This is
achieved by repeat aspiration and dispensing of the solvent prior to the final
aspirate step. This will prevent the dripping that will occur after aspirating a
volatile solvent is aspirated due to partial pressure exerted by the gas phase of
the liquid in the head space.
2.
Post-aspiration air gaps can also help minimized dripping of volatile liquids.
3.
Aspirate and dispense speed options compensate for the resistance-to-flow
that viscous liquids have. Slow pipetting of a viscous liquid will improve
accuracy.
4.
Slow tip withdraws can compensate for liquids with high wetting properties
(liquids that tend to stick to the outside of the tip).
5.
Tip-touch option can remove drops of liquids with high surface tension that
cling to the end of the tip.
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Tomtec
Section XII
HEAD REMOVAL, INSTALLATION AND
ALIGNMENT
Tools Required: # 2 Phillips screwdriver, #2 flat blade screwdriver or 1/4 inch
nut driver, 1/8 Inch Allen wrench, 3/16 Inch Allen wrench, Refer to Figs: 111-22
through 111-33
Procedure: Shut off power.
Take off the top cover by removing the ten Phillips head screws.
1.0 TOP
VIEW
BACK
Access to 3/8 inch
alien head draw bolts
that secure Tip and
Seal Assemblies for
96sv and 384 Heads
Top cover
Open the front
bottom left to snap
side, then pulling it
be able to be
panel by pulling on the
it open from the left
out straight. It will then
swung to the right.
If the head is a
450ul backfill first insure
there is no water in the head, bottle or hoses. Then loosen the clamp holding the hose on to
the fitting on the back left over the backplane and remove the hose.
Loosen the four hold down screws and pull the head forward enough so that you can
reach the two connectors on the top left of the backplane. Once these are disconnected, lift
the head up and forward, being careful not to damage the tips if they are still attached. You
may want to remove the connectors first. To do this, loosen the two forward captive screws
that secure the B Q96 control board mounting plate all the way out. Loosen the two back
ones enough to pivot the plate and then tilt the plate up in front enough to access the two
connectors. Place the head in the storage stand. Never stand the head on the tips or
place it where the tips may be damaged.
178
Tomtec
HEAD REMOVAL, INSTALLATION & ALIGNMENT
CONTINUED
2.0 TOP BACK VIEW
Hold down bolts (4)
Jacking
screws (4)
Jacking
Screws
24VDC
interlock
179
Tomtec
Install the desired head, holding it while putting on the two connectors, then set it in
its proper location. If it's the backfill head, connect the hose.
Screw in the four Allen hold down screws part way. Now it is necessary to align the
head to the stage.
Close the front panel and turn on the power. The stage needs 24VDC to work and
the front door interlock shuts it off.
Initialize the Quadra 4.
Put a nest with a 384 well plate on the station over the stage. The 384 well plate is
used for the alignment of either 96 or 384 well heads.
Using the utility screen, raise the stage to a height so that the tips are just slightly
over the wells. Be careful in setting the height so that you avoid damaging the tips.
Back out all four jacking screws.
If the tips are not level across the plate, use the jacking screws to adjust the head. If
any adjustment is made with the jacking screws, be sure that all jacking screws
make contact before completing the adjustment. Tighten the hold down bolts and
confirm that the tips are still level with reference to the stage. Loosen the hold down
bolts and continue.
For a 96 SV or 450jal head, move the head laterally to align the tips the center of the
cross hairs on a 384 well plate. The cross hairs are the plastic between four wells of
a 384 well plate that correspond to a single well in a 96 well plate. For a 384 head,
move the head laterally to align the tips to the centers of the wells.
Confirm that the head will still index 96 to 384 without the tips hitting the plate.
Insure that straight tips are used on the 450µL head.
Tighten the hold down bolts then recheck the alignment and indexing.
Secure the B Q96 control board mounting plate, if necessary, and install the top
cover.
NOTE: The software will configure to the head installed.
180
Tomtec
3.0 HEAD INSTALLED
Q96 B board
mounting plate
Hose Fitting
Captive screw (4)
Electrical
Connectors
(2)
3/8 inch alien
head draw bolts
181
Tomtec
182
Tomtec
4.0 450µL BACKFILL HEAD C020SS0-023
BACK VIEW
Light switch
(Pinch tubes
Tip shuck
plate
Tip load light
switch pin
assembly
183
Tip pins
(96)
Tomtec
184
Tomtec
7.0 350 60µL HEAD C0S0S691
BACK
VIEW Fig.
Initialization
flag
FRONT
VIEW Fig.
111-31
Interrupter
light switch
Tip on snap
switch
assembly
185
Tip and
Seal
Assembly
(384-104)
Note that
Pistons
are inserted into
Tip
and Seal
Assembly
Tomtec
8.0 384SV TIP AND SEAL ASSEMBLY [384-104]
186
Tomtec
Section XIV
ROUTINE & PREVENTIVE MAINTENANCE BY
OPERATOR
Keep the tops of the stages, stacker, and shuttle clean on a continual basis.
Clean any spilled liquid promptly so it will not dry on moving parts.
1.0
LUBRICATION KIT [196-107]
Lube the tip pin "0" rings daily.
(Applies to the backfill head only) Do this more often if the tips are changed
frequently. (Lubrication Kit, 196-107) (Lubrication Kit Refill, 196-107-1)
Open the "pinch tubes" before powering down the instrument. (Applies to the
backfill head only) Leaving the pinch tubes closed for extended periods of time may
result in damage to the head and impaired pipetting action. It may also void any
warranty or service agreement.
Insure plates are stacked properly. Before starting a program, check for
misaligned plates in the stacker cassettes. Are they properly nested onto each
other? Do the bar code labels overlap the top of the plate, and if so, will they cause
the plates to stick to each other?
Use the follower plates. When no additional plates are to be loaded into the
cassette, add the follower plate. This will add additional weight to the stack and
insure proper loading of the last plate onto the nest.
187
Tomtec
2.0
Cleaning the pipettor tips. (Applies to the 96SV and 384 heads)
While using your Tomtec SV pipettor, wash the tips using the ultrasonic
cleaner by performing the mix function while the tips are immersed in the
flowing bath. The wash volume should exceed the transfer volume by at least 5
µL.
At the end of the day's use, perform the wash function again but for a
minimum of five minutes, using a mix volume of 55ul with a 5ul air gap drawn
first. Then shuck the tip assembly and perform a piston wash for a minimum of
five minutes. Your pipettor is now ready for the next day's use.
If you should find that the above procedure is not adequate for your
level or type of usage, you may consider the following additional
procedure:
After removing the tip assembly, place the assembly upside down in an
ultrasonic bath capable of submerging the entire assembly. The bath should
contain only De-ionized water. Sonicate the assembly for a minimum of 10
minutes. Remove the assembly and allow to dry over night, or dry it by blowing
clean compressed air through the tips.
If you should find that the small orifice at the bottom end of a tip remains
clogged after careful cleaning, you may use the tip orifice cleaner (Catalog No.
020647) equipped with a specially designed bit to remove the clog. One holder
and two bits are provided with each device. To avoid breaking the bit, we
suggest it be placed in the holder so that no more than 1/2 inch is exposed.
Never insert anything into the top of the tip assembly or damage to the
pipetting seal will result.
188
Tomtec
FUSE REPLACEMENT
Turn Main Power switch off. Disconnect power cord from the power entry module located on the
back of the Quadra. Remove the fuse holder located at the top of the power entry module by
pulling outward on the fuse holder's lower tab. With the fuse holder out of the power entry module,
remove the fuses.
Replace fuses according to the following chart:
Return the fuse holder to its position in the power entry module by pushing in until it is seated. Reconnect the power cord into the power entry module. Turn Main Power switch on.
189
Tomtec
WARRANTY
Tomtec Inc. guarantees the Quadra 4 against defects in materials and workmanship.
Defective material will be replaced at no charge for a period of up to one-year following
shipment. Labor required for warranty related repair will be done at no charge for one
year, providing the equipment is returned to our factory for repair. The cost of
transportation both ways must be paid by the customer. This warranty is valid
providing the equipment is utilized within the guidelines of the operation manual.
Failures due to misuse or neglect are not covered by this warranty.
This warranty is exclusive and is in lieu of other warranties, whether written, oral or
implied, including the warranty of merchantability and of fitness for any particular
purpose. Tomtec's liability is, in all cases, limited to the replacement price of its
product. Tomtec shall not be liable for any other damages, whether consequential,
indirect, and incidental, arising from the sale and use of its products.
Note that disabling the screen saver will void the warranty on the front panel display.
RETURN POLICY
It is the responsibility of the user to determine the suitability of the product for
specific application. While defective products will be replaced without charge if
promptly returned, no liability is assumed beyond such replacement. Authorization
for return must be received from Tomtec, Inc. before returning any equipment
for inspection or warranty repair. A signed, completed "Decontamination Form"
must accompany all returns. This is to verify to our employees that the unit is
biologically and radioactively safe to work on.
190
Tomtec
QUADRA 4
DECONTAMINATION FORM
In order to protect personnel involved in the service and repair of instruments, one
must ensure that risk factors hazardous to health (Biological-infectious material or
Radioactive isotope) are removed from the equipment in question.
This form must be signed and accompany the Tomtec instrument when it is returned
for service or repair. If not, the service department may be unable to perform service
on the instrument.
Model:_________________________ Serial Number:_________________________
Type of Contamination:
Method of Decontamination:
I confirm that the instrument listed above has been submitted to an appropriate process of
decontamination at this facility before being shipped to Tomtec, Inc.
Name of Institution: __________________________________________________________
Institution Address: ___________________________________________________________
Date:________ Signature: ______________________________________________________
Print Name:__________________________________________
Title:________________________________________________
NOTE: In all cases, equipment must be emptied of all fluids, prior to returning to Tomtec.
TOMTEC INC. 1000 Sherman Avenue Hamden, CT 06514 USA Phone 203-281-6790 Fax
203-248-5724
191
Tomtec
IF YOU NEED HELP...
If you have any questions regarding this manual or the service and maintenance of any Tomtec
equipment, please contact the Tomtec Inc. Service Department. Please have your model
number and serial number available should the service technician request it.
Our phone and fax numbers in the United States:
Phone: 203-281-6790 Fax: 203-248-5724 www.tomtec.com
Our shipping address within the USA is:
Tomtec Inc.
1000 Sherman Avenue
Hamden, CT. 06514 USA
Our shipping address for International use is:
CONSIGNED TO:
Tomtec Inc.
C/o Allstates World Cargo JFK International Airport Notify on arrival @ (908) 6240505 Fax: (908) 206-9701.
Toll Free Number for USA & Canada:
(877) TOMTEC3
Phone Number for Local Sales & Service:
(203) 281-3683
Tomtec offers distribution and service in many countries throughout the
world. Please see our 'Worldwide Distributors' section for a representative
in your area. Please contact Tomtec me, USA (between the hours of 8:30
am - 4:30 pm EST) if you need any technical support or have any questions
or concerns about any Tomtec product.
Note: Be sure to receive authorization from a Tomtec Service Representative before returning machines for
service or repair. These returns MUST be accompanied by a completed, signed decontamination form.
192
Tomtec

QUADRA 4 Manual Rev-E2

Transcription

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