IsoFlux System - Users Guide (630-0126 Rev C)

IsoFlux™ System User’s Guide Doc. No. 630-­‐0126 Rev C Fluxion Biosciences www.fluxionbio.com Fluxion Biosciences IsoFlux™ System CONTACT INFORMATION: Fluxion Biosciences Inc. 385 Oyster Point Blvd. #3 South San Francisco, CA 94080 Phone: +1 (650) 241-­‐4777 Toll Free (USA): +1 (866) 266-­‐8380 FAX: +1 (650) 873-­‐3665 Web: www.fluxionbio.com Sales [email protected] Technical [email protected] General [email protected] TECHNICAL SUPPORT: Fluxion offers technical support for the IsoFlux System by phone, email, and through service contracts. When contacting Fluxion, please identify the Serial Number of your product. If you purchased your Fluxion product through an authorized distributor, you can contact your service representative for support issues. PRODUCT RETURNS: Please do not return any product to Fluxion without first obtaining an RMA # from a Fluxion representative. 630-­‐0126 Rev C Page 2 Fluxion Biosciences IsoFlux™ System Declaration of Conformance Fluxion Biosciences 385 Oyster Point Blvd, Unit 3 South San Francisco, CA 94080 (650) 241-­‐4777 www.fluxionbio.com Product: IsoFlux™ System Model No.: 950-­‐0100 Serial No: <As marked on unit> The undersigned hereby declares, on behalf of Fluxion Biosciences, Inc. of South San Francisco, CA, that the above-­‐referenced product, to which this declaration relates, is in conformity with the provisions of: EU Directive: EU Standard: 2004/108/EC Electromagnetic Compatibility Testing EN 61326-­‐1 Class A testing for Lab/Industrial Environment 2006/95/EC Safety Testing EN 61010-­‐1 3rd Edition The Technical Construction File required by this Directive is maintained at the corporate headquarters of Fluxion Biosciences, Inc., 385 Oyster Point Blvd. South San Francisco, CA. The product herewith complies with the above-­‐mentioned provisions of normative European Council Directives and carries the CE marking accordingly. Signature on file Steve Kushman, P.E. Director of Manufacturing 630-­‐0126 Rev C Page 3 Fluxion Biosciences IsoFlux™ System IsoFlux System User’s Guide Table of Contents Section Page 1. Safety and Site Requirements……………………………................................... 5 1.1. Safety considerations 1.2. Power requirements 1.3. Bench space 1.4. Environmental considerations 2. Instrument Installation………………………………………………………………………… 7 2.1. Unpacking and placing the system 2.2. Making electrical connection 2.3. System overview 2.4. Powering system 3. User Interface Overview………………………………………………………………………. 11 3.1. Touch screen layout 3.2. Protocol selection 3.3. Settings 3.4. Power off and standby 3.5. Updating software and exporting logs 3.6. Error screens 4. Running Experiments…………………………………………………………………………... 17 4.1. Selecting a protocol 4.2. Priming 4.3. Loading cartridge after sample has been added 4.4. Removing cartridge after assay is complete 4.5. Running separation using previously primed cartridges 5. Cleaning and Maintenance………………………………………………………………..… 25 6. Technical Support Contacts…………………………………………………………………. 26 7. Warranty Information………………………………………………………………………. 27 630-­‐0126 Rev C Page 4 Fluxion Biosciences IsoFlux™ System 1. SAFETY AND SITE REQUIREMENTS 1.1. Safety considerations The following symbols are used in the manual: CAUTION
Calls attention to a procedure, which, if not correctly executed, could result in personal injury or damage to the equipment. Do not proceed beyond a "CAUTION" sign until the indicated conditions are fully understood and met. Calls attention to important information. Read this information before continuing. CAUTION
If the IsoFlux system is used in a manner not specified by Fluxion Biosciences, the protection provided by the equipment may be impaired. To prevent injury, always observe good laboratory practices when you handle reagents, blood, or operate the system. Know the physical and chemical properties of the reagents you use. Refer to the Material Safety Data Sheets for the reagents in use. Follow a regulated, approved waste disposal program. For instructions on sample collection, processing, IsoFlux consumable kit use, and downstream analysis, refer to the specific Instructions for Use from the consumable kit. 630-­‐0126 Rev C Page 5 Fluxion Biosciences IsoFlux™ System 1.2. Power requirements The instrument is powered using the included power cord. Verify that the supplied cord is correct for the installation location. Only the universal adapter and power cord supplied with the instrument should be used. For safe operation, the system must be connected to a grounded 3-­‐prong outlet, where the third prong (round for US outlets) is connected to protective earth ground. Never use a “ground lift adaptor” between the product’s plug and the wall receptacle. Failure to connect to a protective earth ground may result in electric shock, fire, personal injury and death. The system permits operation from any line voltage with the following requirements: Voltage: 100-­‐240 VAC Power: 4-­‐8A Frequency: 50-­‐60 Hz. For performance meeting the CE Guidelines for Surge Resistance of 220/50 or similar AC inputs, use of an external surge suppressor with the following specifications is recommended: Surge Suppression Rating: Transient Response Time: Line Noise Suppression Rating: 1020 Joules Instantaneous > 75 dB @ 1 MHz 1.3. Bench space The Controller requires bench space 18” (45cm) wide x 28” (70cm) deep, and 24” (60cm) tall. The instrument weighs approximately 130 lbs (52 kg). Cartridge loading drawers extend an additional 6” (15cm) from the front of the unit during cartridge loading/unloading and that space must remain unobstructed. 1.4. Environmental considerations The user is responsible for operating the IsoFlux System in a safe environment. The instrument is designed to operate in an environment with ambient temperatures between 20-­‐30 °C (68-­‐86°F) and humidity <95%, non-­‐condensing. The IsoFlux System is not designed to be spark-­‐proof and should not be used in the presence of flammable vapors, gases, or gas mixtures. 630-­‐0126 Rev C Page 6 Fluxion Biosciences IsoFlux™ System 2. INSTRUMENT INSTALLATION To install the system, follow these steps in order: 2.1 Unpack the system and place on lab bench 2.2 Connect power 2.3 Power on system and initiate start-­‐up sequence 2.1. Unpacking and inspecting the system To unpack the IsoFlux system: • Inspect the shipping cartons for damage or evidence of mishandling. If damage or mishandling is evident, notify the carrier before opening the cartons. • Cut the tape and open the flaps on the top of the IsoFlux shipping box. Remove the outer shipping box. • Remove the foam packing material from the top of the IsoFlux unit. CAUTION
• Two people are required to carefully lift the IsoFlux system out of the box. Grasp beneath the sides of the instrument. A third person may be necessary to help slide the carton and foam inserts off the instrument. DO NOT attempt to lift the system by yourself. • Place the instrument on a lab bench in a location with convenient access to power. 630-­‐0126 Rev C Page 7 Fluxion Biosciences IsoFlux™ System Check the contents against the parts list in table to confirm that all items are included. Table 1: Parts List for IsoFlux Shipping Container # 1 2 3 Item IsoFlux System Power Cord User Guide Description Benchtop system for rare cell enrichment Input: 100-­‐240V 50-­‐60Hz Instructions for use Qty 1 1 1 2.2. Setting up electrical connection On/Off switch Power Supply 1. Confirm that Power switch is in the Off position (‘O’ symbol depressed). 2. Connect instrument side of power cord to instrument in the receptacle below the power switch. Ensure that connection is fully engaged. 3. Connect the cord to a power outlet. 630-­‐0126 Rev C Page 8 Fluxion Biosciences IsoFlux™ System 2.3. IsoFlux System components Touchscreen display Control panel: USB on left Standby button on right Sample tray Tray door Overflow drawer 630-­‐0126 Rev C Page 9 Fluxion Biosciences IsoFlux™ System 2.4. Power system on and initiate start-­‐up sequence 1. Turn Power switch on back of unit to On position (top of switched, marked ‘I’, depressed). 2. The Standby button on the front panel will now flash orange. This means power is on but the system has not yet started. Press the Standby button briefly. It will now change to green and the system will begin the boot-­‐up procedure. Boot-­‐up will take approximately 4 minutes, during which the following screen will be displayed: 3. Watch the touch screen to confirm that system powers on correctly. The Home screen shown below will be displayed. The system is now ready for use. 630-­‐0126 Rev C Page 10 Fluxion Biosciences 3. USER INTERFACE OVERVIEW 3.1. Touchscreen layout 630-­‐0126 Rev C IsoFlux™ System Global Navigation buttons Bright-­‐ button active Dim-­‐ button inactive Home Settings Back Screen-­‐specific buttons Information bar: contains information relevant to screen and assay status Page 11 Fluxion Biosciences IsoFlux™ System 3.2. Protocol Selection Screen The protocol screen allows the user to select any of the available isolation protocols that have been loaded. From the Home screen press the ‘SELECT PROTOCOL’ button to access the Protocol screen. The first four available protocols will be displayed. Additional protocols, if available, can be viewed by pressing the button labeled “More”. Press the desired protocol to make it the active version. Any runs started from the Home screen will use the active protocol. 630-­‐0126 Rev C Page 12 Fluxion Biosciences IsoFlux™ System 3.3. Settings Turn audible button press alerts on/off Export all logged runs and events to USB Install new software (provided on USB from Fluxion) Shut down system 3.4. Power Off and Standby Pressing “Power Off” launches this screen. Shutdown turns the system off and would be used for a shutdown period longer than 1-­‐2 days. Sleep places the system in a hibernation mode and turns off the system temperature control and touchscreen to prevent screen burn-­‐in. This option should be used for overnight periods. The Sleep / Shutdown screen can also be accessed by pressing the green power button and holding for 1-­‐2 seconds. While the system is off but wall power is being supplied, the front power button will glow orange. As described in Section 2.4, power can be restored by pressing the power button on the front of the instrument and holding for 1-­‐2 seconds. The touch panel should light up when the system has turned on. Recovery from sleep mode will be immediate, while bootup from a complete shutdown will take approximately 4 minutes. 630-­‐0126 Rev C Page 13 Fluxion Biosciences IsoFlux™ System If a mechanical error occurs at any time during a run, you will see a message indicating a mechanical error has occurred, and containing a CONTINUE button. Once the user acknowledges the error by pressing CONTINUE, the system will try to reset itself. A mechanical error indicates that either one of the moving parts is stuck or the motion driver is not working properly. If samples were loaded at the time of the mechanical error, the system will retrieve the sample as if it was the end of the run. Therefore, if separation was almost complete, useful sample recovery may result; if early in the separation process, the samples will likely be lost. If a software error occurs, the following screen will be displayed. 630-­‐0126 Rev C Page 14 Fluxion Biosciences IsoFlux™ System Acknowledge the message by pressing OK, and the system will attempt to restart and clear any errors. Any samples present in the system at the time of restart will be presented to the user upon system reboot. Samples may be viable if in the last (wash) stages of a separation protocol. In other cases, if samples were loaded at the time the error occurred, they will be compromised. If either mechanical or software errors happen repeatedly, please contact Fluxion for support. If the system becomes unresponsive, it may be forcibly shut down by either holding down the front power button for over 30 seconds, or by turning off the power switch at the back of the unit. It is not recommended to power down this way unless the system is indeed unresponsive. In particular, before any shipment of the unit a proper shutdown procedure (from the software) needs to be completed in order to protect components during transport. ATTENTION: DO NOT ship the unit without completing a shut down from the touch screen power menu. 3.5. Updating Software and Exporting Run Logs Pressing “Install Software” Launches the Software Update screen. Follow the prompts in the information bar to install new software (supplied by Fluxion only). Pressing “Export Log” will pull up the following screen. Select the desired option and follow the instructions in the information bar. 630-­‐0126 Rev C Page 15 Fluxion Biosciences IsoFlux™ System 3.6. Error Screens The IsoFlux system monitors numerous parameters during a run. If an error is detected the screen will display a graphic indicating a problem. In the screen below, the cartridge in position 2 failed to prime properly and is marked with an ‘X’, while positions 1 and 3 primed properly and are marked with a check indicating success. 630-­‐0126 Rev C Page 16 Fluxion Biosciences IsoFlux™ System 4. RUNNING EXPERIMENTS A general outline is provided here for a typical IsoFlux assay. Detailed instructions for sample collection, processing, and kit use can be found in the IsoFlux enrichment kit Instructions for Use. Additional assay protocols and technical information can be found at www.fluxionbio.com. For further support, please contact Fluxion Biosciences ([email protected]). A typical IsoFlux run consists of the following 4 steps: 1. Select a protocol 2. Prime the cartridges prior to loading samples in them 3. Loading samples into the cartridges (see IsoFlux kit Instructions for Use for details on sample processing) 4. Removal of cartridges after the run is complete 4.1. Select a protocol All samples during a run use the same protocol. Go to the Select Protocol screen if you wish to change to a different protocol. If no change is made, the same protocol from the previous run will be loaded. The selected protocol name is displayed in the information bar on the Home screen. Press Run Protocol to start. 630-­‐0126 Rev C Page 17 Fluxion Biosciences IsoFlux™ System 4.2. Prime the cartridges Pressing Run Protocol will display the sample selection screen. Choose the appropriate number of samples in the run, from one to four. The following screen is displayed (3 sample run shown, as an example): 630-­‐0126 Rev C Page 18 Fluxion Biosciences IsoFlux™ System Load the cartridges with buffer as instructed in the information bar and press “PRIME” when complete. For more details on working with IsoFlux cartridges, see the appropriate IsoFlux kit Instructions for Use. The priming step takes several minutes. After the priming is complete the following screen is displayed: 630-­‐0126 Rev C Page 19 Fluxion Biosciences IsoFlux™ System If any cartridges did not prime properly, they will be displayed with an ‘X’ across the sample as shown here: If any cartridges do not prime properly, the experiment will continue for the successfully primed consumables only. The user should leave the faulty consumable on the tray and proceed with sample loading for the successfully primed consumables only. No re-­‐priming operations are provided for, or likely to be successful for that particular consumable. Priming operations should be successful a very high percentage of the time; if this issue is encountered, please contact Fluxion support. Do not press ‘READY TO LOAD SAMPLES’ until all samples are fully prepared and ready for immediate transfer into the cartridge. The cartridge is maintained at a reduced temperature when inside the system, and extended exposure outside the system will degrade isolation and recovery of the target cells 4.3. Load cartridge after sample mixed with beads has been added Press ‘READY TO LOAD SAMPLES’ when ready. The left side door will open first. Load one or two samples, as appropriate, into their slots and press ‘LOAD’. After the left door closes, the right door, if 3 or more samples are being processed, will open. Load samples in cartridges 3 and 4 and press ‘LOAD’. Only load samples in consumables that have successfully completed the prime phase If the system detects any problems with the samples, such as insufficient volume, an error message will be generated and the tray will open. Correct the problem and reload (see the IsoFlux kit Instructions for Use as well as the troubleshooting guide for more information). 630-­‐0126 Rev C Page 20 Fluxion Biosciences IsoFlux™ System 4.4. Remove cartridge after sample has been processed The isolation and recovery of the target cells takes 30-­‐45 minutes. If any consumables have failed the priming or separation phases for any reason, they will be indicated by a red color (see section 4.3). Once the run is completed the following screen is displayed: 630-­‐0126 Rev C Page 21 Fluxion Biosciences IsoFlux™ System Do not press ‘EXTRACT SAMPLES’ unless you are ready for the next step in the assay or are ready to store the cells. Target cells will degrade rapidly in ambient conditions. Press ‘EXTRACT SAMPLES’ to open the sample drawers. The following screen appears: The timer is a reminder to remove the samples quickly; target cells will begin to degrade in ambient conditions and time is of the essence. We recommend moving the cells to processing or storage within 5 minutes. 4.5. Running separation using previously primed cartridges In certain cases it may be necessary to run the system using previously primed cartridges. For this eventuality, there is a protocol entitled ‘No prime protocol’ and the priming step is replaced by a ‘dummy prime’ that is short and doesn’t alter the already primed state of the cartridges. Please note that this mode of operation is not ideal and should only be followed when unforeseen circumstances cause a normal run not to be feasible. As an example, on Day 1 cartridges are primed, but the patient blood sample is unavailable to complete that run. In this case the cartridges are then kept primed until the next day, when the blood sample 630-­‐0126 Rev C Page 22 Fluxion Biosciences IsoFlux™ System becomes available. Note: DO NOT allow the channel to dry as this will result in crystal formation that may clog the channel. In this scenario, the following steps should be followed: Let’s assume that priming for 3 cartridges went OK, and the user is presented with the following screen: Press ‘ABORT’ and remove all cartridges from the carriages. Manually and gently remove the CellSpot coupons from the low volume holder or the 0.6mL microfuge tube AND place each one into the corresponding CellSpot wells (well number 4) After the primed cartridges are removed from the instrument, make sure that ALL CellSpot coupons are in the correct position, namely in the CellSpot well. CellSpot coupons may have to be manually removed from the low volume holder and re-­‐inserted into the CellSpot well, as shown below. If coupons are not replaced, the instrument will assume they are there and continue the run without priming. This will result in overflow and possible sticking of the magnet shaft. The figure below shows a CellSpot coupon in the improper location (inserted into the low volume holder, A), and the process for manually removing the CellSpot coupon (B) and replacing it into the proper location, at the bottom of the CellSpot well (well number 4, C). 630-­‐0126 Rev C Page 23 Fluxion Biosciences IsoFlux™ System After the coupons have all been placed in the proper locations in their respective wells, the cartridges can be stored for up to 48 hours at 4°C, but should be used as soon as possible. Before running the cartridges, inspect and make sure that the wells are still hydrated. During long term storage, the microfluidic channels may become clogged if they dry out and the resulting salt crystals remain inside the channel. Finally, when the sample is ready to run (i.e. the next day), the user can load the protocol entitled ‘No prime protocol’ and run through the on-­‐screen prompts. 630-­‐0126 Rev C Page 24 Fluxion Biosciences IsoFlux™ System 5. CLEANING AND MAINTENANCE Inspect the instrument, including the trays and drip pan beneath it, on a daily basis. To inspect the drip tray, start a protocol to open the drawers. Check the white plate that sits below the tray when the tray is inside the instrument. If there are any signs of fluid spills on the tray, wash it with a mild bleach solution. If there are signs of spills in the drip pan wash it with a mild bleach solution. The bleach will drain into the overflow drawer (see below). Allow 30 minutes to drain completely, then remove the overflow drawer and properly dispose of the waste. On a weekly basis, saturate a paper towel with 70% isopropanol or ethanol and wipe the tray thoroughly. Decontamination time with 70% alcohol is 30 minutes of wet exposure to the biocide. Allow the system to air dry following decontamination. 630-­‐0126 Rev C Page 25 Fluxion Biosciences IsoFlux™ System 6. TECHNICAL SUPPORT CONTACTS For IsoFlux Technical Support, please contact: Phone: +1 (650) 241-­‐4777 Toll Free (USA): +1 (866) 266-­‐8380 FAX: +1 (650) 873-­‐3665 Web: www.fluxionbio.com Sales [email protected] Technical [email protected] General [email protected] Please do not return products to Fluxion without first obtaining an RMA#. Include this RMA # on the shipping address: Fluxion Biosciences Inc. Attention: Product Returns (RMA# XXXX) 385 Oyster Point Blvd. #3 South San Francisco, CA 94080 630-­‐0126 Rev C Page 26 Fluxion Biosciences IsoFlux™ System 7. Warranty Information Fluxion Biosciences warrants IsoFlux instruments against defects in materials and workmanship for a period of one year from date of arrival to the customer. During the warranty period, Fluxion will, at its option, repair or replace products that prove to be defective. Fluxion warrants that its software and firmware designed for use with a CPU will execute its programming instructions when properly installed on that CPU. Fluxion does not warrant that the operation of the CPU, or software, or firmware will be uninterrupted or error-­‐free. Warranty service Warranty service is performed at no charge and at Fluxion's option in one of the following methods: • A service representative is dispatched to the customer’s facility. • The product is returned and repaired at a Fluxion repair facility. • Replacement parts with appropriate installation instructions are sent to the customer. The foregoing warranty shall not apply to defects resulting from: • Improper or inadequate maintenance, adjustment, calibration • Buyer-­‐supplied software, hardware, or consumables • Unauthorized modification or misuse • Operation outside of the environmental and electrical specifications for the system • Improper site preparation THE WARANTY SET FORTH IS EXCLUSIVE AND NO OTHER WARRANTY, WHETHER WRITTEN OR ORAL, IS EXPRESSED OR IMPLIED. FLUXION SPECIFICALLY DISCLAIMS THE IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE. Limitation of Remedies and Liability THE REMEDIES PROVIDED HEREIN ARE BUYER’S SOLE AND EXCLUSIVE REMEDIES. IN NO EVENT SHALL FLUXION BE LIABLE FOR DIRECT, INDIRECT, SPECIAL, INCIDENTAL, OR CONSEQUENTIAL DAMAGES (INCLUDING LOSS OF PROFITS) WHETHER BASED ON CONTRACT, TORT OR ANY OTHER LEGAL THEORY. 630-­‐0126 Rev C Page 27