TM-Sprayer™ Tissue MALDI Sample Preparation System Technical Note

TM-Sprayer™
Tissue MALDI Sample Preparation System
Technical Note
Optimization of Automated Matrix Spraying for MALDI Imaging
The TM-Sprayer™ System is an automated MALDI plate matrix deposition system offering high
reproducibility and superior data quality.
TM-Sprayer™ is an easy-to-use, versatile
spraying system that provides an automated
process for the preparation of MALDI
plates. The patented technology1 of the TMSprayer™ guarantees a very uniform and
consistent coating crucial for reliable
imaging.
Automated Matrix Deposition
Techniques for spray coating have utilized
manual methods of matrix deposition, and
are subject to variations dependent on the
individual
preparing
the
sample.
Standardization of the matrix deposition
provides a uniform and reproducible
methodology.
In the TM-Sprayer, a constant flow of
heated sheath gas is delivered conjointly
with the matrix spray which results in two
major benefits:
The unique design of the TM-Sprayer™
spray nozzle creates a fine solvent mist that
is transported by a heated inert gas and
quickly evaporates upon contact with the
MALDI plates. Spray characteristics are
easily adjustable via our intuitive Operator
Interface. Users can select from a large
selection of spray pattern designs and plate
types to meet the requirements of their
application.
1
Applicable patent 5,772,964
•
Very rapid evaporation for minimal
analyte delocalization.
•
Highly efficient co-crystallization for
superior mass spec signal.
Coupled with the benefit of faster sample
preparation, the TM-Sprayer™ delivers the
automated matrix deposition solution
required to keep up with the high speed of
the latest MALDI Mass Spectrometers.
Large choice of Matrix
The design of the matrix flow path provides
a great flexibility in solvent choices. The
TM-Sprayer™ permits processing of up to
1.0ml/min of matrix solution. Flow rates are
controlled through an auxiliary pump.
Consistent Matrix Spraying
The temperature control technology used for
the patented nozzle of the TM-Sprayer™
ensures very stable and reproducible spray
characteristics, across a wide range of
matrix solvent.
Intuitive User Interface
•
•
•
Adjustable Method Parameters
Nozzle height above target, temperature of
matrix/solvent mix, temperature of gas and
forward speed of cross table are easily
controlled to result in proper application
wetness, optimal resolution and extraction
performance.
High Quality Robotics
The precise robotics of the TM-Sprayer™
deliver a smooth and constant-speed linear
displacement of the MALDI target along the
X-Y axis, which results in a uniform matrix
deposition. Hot-spots often associated with
manual techniques are eliminated.
Easy Method Development
The TM-Sprayer can save an unlimited
number of methods, which makes it easy to
access optimal
parameters for
each sample type
of MALDI plate,
matrix chemistry
or types of tissue
samples.
•
•
Microsoft Windows™ Operator
Interface
Single view User Interface
Graphical Selection of the desired
deposition areas
Linear offset tracks or an alternating
90°
rotation
during
matrix
application
Save and Load routine sample
programs
Large selection of Spray Pattern Design
Select from serpentine to vertical or
horizontal line patterns.
Custom configurable MALDI plate
interface and multi-vendor plate holder
Create new plate types using the simple
plate configuration tool.
SPRAY AUTOMATION IMPROVES SAMPLE PREPARATION AND REPRODUCIBILITY
Figure 1: Improved reproducibility
Figure 2: Improved data quality
1411
Day
496
856
1411
Manual
Spray
Manual
Spray
TM
Sprayer™
TM
Sprayer™
Day
Less Protein
Delocalization
Fewer Hot Spots
Sharper images
All of the above images were acquired using 150 micron resolution. Samples were mouse brain sections. Manual spraying was performed using a glass Venturi
sprayer. Details of automated spraying shown on right panel. Reproducibility studies were performed on different days, while data quality was evaluated using
serial sections.
High resolution MALDI mass spectrometric image of a mouse brain
Method: Normal mouse brain was sectioned at 12 micron thickness and thaw mounted onto a metal MALDI target. The sections were
dehydrated using ethanol washing and dried in a desiccated environment. Matrix was applied with the TM Sprayer™ using the following
parameters:
142.5º C
Sinapinic Acid 5mg/mL
Block Temperature:
Matrix:
300 mm/min
50% Acetonitrile in 0.1% TFA
Stage Speed:
240 μL/min
10 psi
Matrix Flow Rate:
Air Pressure:
3 mm
Line Spacing:
40 mm
Nozzle Distance:
Figure 3: After drying, the sample is imaged at 75 micron resolution and visualized using BioMap™ (Novartis)
1411
752
1519
6219
4970
671
5969
499
14166 (r) + 15198 (g)
14166 (r) + 4970 (g)
14166 (r) + 6219 (g)
14166 (r) + 4990 (g)
Technical Characteristics
TM-Sprayer™ nozzle design ensures a
reproducible matrix from one sample to the
next.
Figure 4
Temperatu
Temperature
Contr
Control
l
N2
Sheath
G
block
heater
h t
• Sheath Gas – Ambient to 200°C (+/- 2°C),
software selected
• Collection Unit – X-Y Robotic Stage,
fixed nozzle
• Gas Supply Sheath gas flow 5-15.5
liter/min
• Electrical 120 VAC, 60 Hz or 220 VAC,
50Hz: 2.5A fused
• Dimensions/Weight 42x34x31 cm, 14kg
Preheat
capillary
coiled in block
it
LEAP Technologies offers a complete line
of sample preparation platforms for
MALDI imaging
matrix
spray
TM-iD™ Vision Enabled
Liquid Dispensing for
MALDI Spotting.
2D iDx™ Automated
Gel Processing System
Minimal Maintenance and Easy Service
All key components are easily accessible for
quick adjustment or maintenance.
Figure 5
MALDI Spotter kit
for HTS PAL
Acknowledgements
Protein Discovery, Knoxville, TN
Additional Information
TM-Sprayer™ is available worldwide from
LEAP Technologies, Inc.
To request further information please call
LEAP Technologies at +1-919-929-8814
Contact:
TM-Sprayer™ Specifications
• Deposition – Spray deposition in linear or
serpentine modes with variables offsets.
Spray
patterns
by
alternating
perpendicular paths for even coatings
• Nozzle Flow – 50 to 1000µl/min
Alain Creissen
International Sales Manager
[email protected]
LEAP Technologies
PO Box 969
Carrboro, NC 27510
+1-919-929-8814
[email protected]
www.leaptec.com