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iQue

Hardware Manual: Part Number 10554 Rev C., Getting to Know the iQue

Fluorescence Channels

The following table lists the configurations of the four fluorescent channels of the iQue Screener in its default laser

and filter configuration along with examples of some flourophores that can be used with the iQue. Note that this is

not meant to be a comprehensive list of all available fluorophores that work with the iQue. Also note that alternate

laser and filter configurations are possible using the Selectable Detector Sets optional feature.

Channel Position

Filter

Example Fluorophores

FL1

533/30

•

FITC

•

GFP

•

eGFP

•

YFP

•

Alexa Fluor

488

•

DiO

•

ToTo

•

YoYo

•

Bodipy

•

CFSE

FL2

585/40

•

PE-Texas Red

•

DSRED

•

Cy3

•

DiL

FL3

670LP

•

7-AAD

•

PerCP

•

PerCP-Cy5.5

•

PerCp-eFluor

710

•

PE-Cy™5

•

PE-Cy7

FL4

675/25

•

APC

•

Alexa Fluor

633/647/670/700

•

DiD

•

ToPro

•

Draq5™

•

Cy5

Summary of Contents for iQue SCREENER

Page 1: ...iQue SCREENER hardware manual October 2014...

Page 2: ......

Page 3: ...1 General Warnings 11 Operator Safety 12 Safety Symbols and Labels 13 Biological Handling Precautions 14 Laser Precautions 14 Operational Precautions 15 Electrical Precautions 15 GETTING TO KNOW THE I...

Page 4: ...he Water Test 36 Interpreting the Water Test 37 Running the 8 Peak Beads Test 37 Running the 6 Peak Beads Test 38 Interpreting the 6 and 8 Peak Bead Tests 39 Daily Shutdown 41 Overview 41 Steps 41 IQU...

Page 5: ...be Required 65 Wetting Procedure 67 Overview 67 Running Ethanol on Fast for 10 minutes 67 Running Water on Fast for 10 minutes 68 Backflush 68 Unclog 68 Managing Plate Models 69 Overview 69 Understand...

Page 6: ...Data 90 110V Power and Data Connection Diagram 91 220V Power and Data Connection Diagram 91 Moving an iQue 93 Overview 93 Referenced Procedures 93 Shutting Down before a Move 93 Moving the Instrument...

Page 7: ...ectronic photographic mechanical or otherwise for any purpose without the express written permission of IntelliCyt Corporation Trademarks IntelliCyt HTFC iQue and ForeCyt are trademarks of IntelliCyt...

Page 8: ...pt the Product AS IS or return the Product and receive a refund of fees paid for the Product 7 Limited Warranties Exclusion of Certain Warranties 7 1 Limited Warranty IntelliCyt warrants that for one...

Page 9: ...not assign any of its rights or obligations under this Agreement without IntelliCyt s prior express written consent which may be granted or withheld at IntelliCyt s sole discretion Any attempted assi...

Page 10: ...TIAL DAMAGES HOWEVER CAUSED AND ON ANY THEORY OF LIABILITY WHETHER FOR BREACH OF CONTRACT TORT INCLUDING NEGLIGENCE OR OTHERWISE ARISING OUT OF OR RELATED TO THIS AGREEMENT INCLUDING BUT NOT LIMITED T...

Page 11: ...5 iQue Hardware Manual Part Number 10554 Rev C Introduction Declaration of Conformity...

Page 12: ...6 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C Introduction...

Page 13: ...ick motion Double click Place the mouse pointer over the item then depress and release the primary mouse button twice in quick succession Drag Place the mouse pointer over the item depress and hold do...

Page 14: ...precaution statements DANGER Indicates an imminently hazardous situation that if not avoided will result in death or serious injury WARNING Indicates a moderately hazardous situation that if not avoid...

Page 15: ...Opt 2 Before contacting us please be prepared to answer the following questions What were you doing when the problem occurred Did you try to follow the steps in this guide but they didn t work If so w...

Page 16: ...10 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C Using This Manual...

Page 17: ...he iQue System the system must be connected to an IntelliCyt approved uninterruptable power supply General Warnings THE iQue SCREENING SYSTEM IS INTENDED FOR GENERAL LABORATORY AND RESEARCH USE ONLY I...

Page 18: ...Take full advantage of all safety features and do not defeat safety interlocks and or sensors Acknowledge and act upon instrument alarms and error messages IF THIS INSTRUMENT IS USED IN A MANNER NOT...

Page 19: ...ening Wear appropriate laboratory attire follow universal laboratory safety protocols and adhere to local regulations Electrical Shock Hazard There is the possibility of electrical shock when the inst...

Page 20: ...Inlet with a tube of HPLC reagent grade filtered distilled water 0 22 m filter or equivalent when the cytometer is not running samples The screening system is programmed for automatic decontamination...

Page 21: ...uding sample tubes and racks Before manually cleaning the system turn off the instrument and unplug the power cord Electrical Precautions Electrical devices pose the risk of an electric shock To reduc...

Page 22: ...16 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C Safety...

Page 23: ...ved uninterruptable power supply UPS The UPS must be plugged into a standard grounded or earthed mains electrical outlet conforming to local codes Non grounded or non earthed mains outlet adaptors mus...

Page 24: ...d Optics Sample Formats iQue Screener 96 and 384 Well Microplates Tubes iQue Screener HD 384 and 1536 Well Microplates Tubes Sample Volumes Sampling Rate 1 L and Higher no dead volume 10 000 Objects S...

Page 25: ...t this is not meant to be a comprehensive list of all available fluorophores that work with the iQue Also note that alternate laser and filter configurations are possible using the Selectable Detector...

Page 26: ...able Gen 2 iQue Contact sensing and piercing probe assembly Fluid stations labeled Buffer Markers Decon Clean and Water 96 and 384 well plate capable iQue HD Gen 2 iQue plus high RPM shaker and advanc...

Page 27: ...iQue Doors Open View iQue terminology 4 26 2013 Bottle Drawer Top Door Detector Cover Sampling Area Door Top Pump Area Sampling Area Connection Area Detector Inlet Shaker Detail View Shaker Deck Gripp...

Page 28: ...the iQue Sampler Detail View Gen 1 Sampler Z Arm Probe Locking Nut Outer Probe Adapter Outer Probe Probe Guide Probe Tip Sampler XY Arm Sampler Detail View Gen 2 Sampler XY Arm Probe Locking Nut Probe...

Page 29: ...23 iQue Hardware Manual Part Number 10554 Rev C Getting to Know the iQue Fluid Station View Gen 1 Fluid Station Detail View Gen 2...

Page 30: ...Pass Through Sampler Tubing Tubing Stop Pump Release Trigger Sample Area Pass Through Pump Rollers Sheath Filter Compression Adjustment Knob Detector Area Detail View iQue terminology 4 26 2013 Flow...

Page 31: ...reen yellow or red that corresponds to the current state of the iQue ready warning and not ready System Status Light The iQue system status is displayed on the hardware and in the ForeCyt software The...

Page 32: ...s not faulted Not Ready The system cannot prime run or clean due to a fault For detailed messages about the system status press the Show Details button on the Controller console Status messages will a...

Page 33: ...other best practices are necessary to maximize instrument performance and to minimize clogs and repairs Please become familiar with these procedures and implement labratory protocols to ensure that th...

Page 34: ...28 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C iQue Operations Cover Page...

Page 35: ...tion and followed by water Fluid Description Preparation Sheath Fluid Part 90078 Used to hydrodynamically focus the samples 1L water 1 bottle IntelliCyt Bacteriostatic Concentrate Decontamination Solu...

Page 36: ...strictly prohibited Filling the Bottles 1 Open the bottle drawer 2 Unplug each bottle by pressing the retaining clip on the bottle tubing connector and firmly pulling up on the plug in connector reta...

Page 37: ...iQue screener is a non pressurized system 10 Carefully close the drawer making sure that the bottle lines are not pinched NOTE Be sure that the bottle connectors are securely plugged in and that the...

Page 38: ...ns is pre set Fluid stations should be filled to approximately 14 mm above the top of the V of the trough 14 mm Gen 2 Trough Fill Line With a Gen 1 probe the probe depth in the fluid stations is user...

Page 39: ...nch the ForeCyt software if it is not already running When prompted to power on the detector click Yes to power it on Alternately click the Power On Detector button in the Controller Console window 3...

Page 40: ...either an improper shutdown or possible fluidics issues such as clogging 5 Perform Daily QC as described in the Daily QC procedure 6 Attach the sampler tubing to the Detector Inlet 7 Click Prime and a...

Page 41: ...QC Samples 1 Prepare the Water Test sample Add 1 mL of 0 22 m filtered distilled water or equivalent to a tube It is important to use clean water for an accurate test 2 Prepare the 8 Peak Beads sampl...

Page 42: ...mplate that matches the bead lot being used NOTE Contact IntelliCyt Tech Support to obtain the latest Daily QC Template for any specific bead lot 4 Give the Daily QC experiment a name and click OK Run...

Page 43: ...orm a Backflush by clicking the Clean button on the controller console choosing the Backflush option and following the wizard 2 Place the 8 Peak Bead sample on the detector inlet 3 Select the 8 Peaks...

Page 44: ...ad sample on the detector inlet 3 Select the 6 Peak Beads tube in the experiment 4 Click Run The experiment will stop after collecting 50 000 events which should take approximately 2 4 minutes 4 If ne...

Page 45: ...ulation is doubled blurred or contains unexpected tails the system may contain an air bubble or other source of fluidic instability Examples of poor scatter for both 6 and 8 Peak Beads Performing a Qu...

Page 46: ...40 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C iQue Operations Daily QC...

Page 47: ...tely 45 minutes to complete Steps 1 Click Clean in the ForeCyt Controller Console window 2 In the Clean wizard select Clean Type Daily Clean and check Shutdown after Clean and then click Next 3 Verify...

Page 48: ...42 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C iQue Operations Daily Shutdown...

Page 49: ...ance and to minimize clogs and repairs Please become familiar with these procedures and implement labratory protocols to ensure that the following best practices are followed Safety Always wear approp...

Page 50: ...44 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C iQue Maintenance Cover Page...

Page 51: ...at startup Daily at startup Daily Clean Daily at shutdown Daily at startup Daily at shutdown Quick Clean As needed As needed Hydrocycle Clean As needed As needed Change Probe and Tubing after 15 hour...

Page 52: ...46 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C iQue Maintenance Recommended Maintenance Schedule...

Page 53: ...e does and when to use it All of the standard cleaning procedures are accessed by clicking the Clean button and using the Clean wizard Accessing the Clean Wizard To perform any standard cleaning opera...

Page 54: ...own procedure will be done after the main cleaning cycle This adds approximately 15 minutes to the clean and powers off the detector Quick Clean The Quick Clean is a short procedure that cleans the de...

Page 55: ...ally performed prior to the normal startup Hydrocycle Clean The Hydrocycle works by starting and stopping the detector fluidics several times in succession This creates pressure changes that are effec...

Page 56: ...50 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C iQue Maintenance Standard Cleaning Procedures...

Page 57: ...edure Required Materials Part Number Description 90659 iQue Probe Tubing Assy for Gen 2 iQue iQue HD 5 Pk 90660 iQue Probe Tubing Assy for Gen 2 iQue iQue HD 10 Pk 90293 iQue Probe Tubing Assy for Gen...

Page 58: ...out of the Detector area 4 Working in the main sampling area draw the tubing through the sampler area pass through Pull firmly for the tubing stop to come through Detach the Old Tubing 1 In the Pump a...

Page 59: ...waste container 1 Lift the probe and tubing assembly out of the upper probe guide 2 Unscrew the brown fitting from the probe holder assembly 3 Pull the inner probe out of the probe holder and discard...

Page 60: ...sitioned across the middle of the rollers Holding gentle tension on the tubing close the pump head locking mechanism Note that the green stopper is pulled fully against the tubing guide on the left an...

Page 61: ...ue Hardware Manual Part Number 10554 Rev C iQue Maintenance Changing the Probe and Tubing 3 Feed the tubing down through the hole leading into the detector area 4 Re attach the tubing to the detector...

Page 62: ...o this run a Daily Clean without the shutdown option Adjusting Pump Head Tension After replacing the probe and tubing it is important to check sample slug formation and adjust the pump head tension if...

Page 63: ...ew one half turn 180 degrees clockwise This should provide enough tension so that the samples move evenly without breaking up and without overly wearing the tubing 9 If this does not improve the quali...

Page 64: ...iQue Hardware Manual iQue Maintenance 58 THIS PAGE INTENTIONALLY LEFT BLANK...

Page 65: ...rmed every two months bi monthly In a heavy usage situation this maintenance may need to be performed more frequently See below for instructions on inspecting parts for wear to determine if fluidics m...

Page 66: ...ctor It is not necessary to shut down the detector before doing this as the optics cover contains an interlock that automatically keeps the system from running when the cover is off 1 Open the lid of...

Page 67: ...h filter on the left of the detector In line Sheath Filter 2 Untighten the Luer lock fittings Remove and dispose of the old filter 3 Remove the new in line sheath filter from its package and write the...

Page 68: ...Detector Peristaltic Tubes 1 Locate the sheath and waste pumps on the top of the detector Sheath Pump Waste Pump 2 Disconnect the Luer locks to the tubing on the outside of the peristaltic pumps by un...

Page 69: ...y from its package and write the current date on the side with a permanent marker 6 Slide one end of the peristaltic tube assembly into the slot on the pump head Route the tubing around and secure the...

Page 70: ...Hardware Manual Part Number 10554 Rev C iQue Maintenance Fluidics Maintenance Impinging Correct Cross Slotted Correct Kinked Correct Examples of improperly and properly installed peristaltic tube ass...

Page 71: ...filters and install them These are the small disk filters Sheath Bottle Filter Decon Bottle Filter Cleaner Bottle Filter 6 Replace the bottle lids top of the fluids and reinstall the bottles in the dr...

Page 72: ...66 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C iQue Maintenance Fluidics Maintenance...

Page 73: ...as been introduced into the system NOTE If this procedure is being used with an instrument that has been out of use for a while and the in line sheath filter has dried out the in line sheath filter wi...

Page 74: ...tor inlet 2 Add another test tube to the experiment and set the Test Tube Time Limit to 10 minutes 3 Run the experiment Backflush 1 Click the Clean button 2 In the Clean wizard choose Clean Type Backf...

Page 75: ...ing sampling This offset is a measure of how far above the deck the probe should go Sampling Height is calculated as follows Deck Height Depth Offset Sampling Height With the Gen 1 probe assembly a si...

Page 76: ...selected By default the system comes with only two plate models listed the Generic 96 Well Plate and Generic 384 Well Plate models Additional plate models may be added to this list for each format No...

Page 77: ...me manufacturer and type for the new plate model 3 Click Save A confirmation prompt will appear for callibrating the plate model Click Yes 4 Follow the instructions in the Plate Model Calibration wiza...

Page 78: ...d and click Calibrate 3 This will bring up the Plate Model Calibration wizard Follow the wizard instructions carefully 4 Place the plate being calibrated on the deck and click Next 5 Use the Vertical...

Page 79: ...wizard instructions carefully 4 Place the plate being calibrated on the deck and click Next 5 The next step in the wizard allows for the configuration of an optional X Y offset See below for discussio...

Page 80: ...om the side or corner of the well The optional X Y offset on a plate model makes this possible To callibrate an X Y offset for a flat bottom plate check the Create Offset Position check box within the...

Page 81: ...e Edit Plate Model window click Make Default 4 A confirmation prompt will appear Click Yes 5 Click Save in the Edit Plate Model screen The Plate Model is now the default and will be used on new Experi...

Page 82: ...76 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C iQue Maintenance Managing Plate Models...

Page 83: ...sing common suspension cell lines IntelliCyt recommends a final cell concentration of between 1 3 x106 cells mL Cell densities that are higher or lower are possible but should be tested and optimized...

Page 84: ...the Rinse section of the Protocol tab check the box to Enable Probe Rinse and set the amount of wells after which a rinse should occur For example to run a rinse after every well choose After every 1...

Page 85: ...within the plate follow these steps 1 In the Rinse section on the Protocol tab check the box to Enable Probe Rinse and set the amount of wells after which a clean should occur For example to run a cl...

Page 86: ...selecting Water for 2000 ms 2 seconds 5 Repeat one last time selecting Buffer for 1000 ms 1 second This will create a standard Decon Clean Rinse mini clean The purpose of the final rinse in Buffer is...

Page 87: ...fall below a certain threshold for longer than expected taking into account shakes and cleans which naturally lower the event rate then the clog detector triggers a warning Click Yes to stop the run i...

Page 88: ...cking Run the system takes well over a minute to begin acquiring data At the end of a run or after clicking Stop the system takes well over a minute to fully stop When running a sample from a tube the...

Page 89: ...y recommended that iQue users contact IntelliCyt Tech Support prior to performing any of these procedures Safety Always wear appropriate safety attire such as protective gloves and eye wear when perfo...

Page 90: ...84 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C Special Topics Cover Page...

Page 91: ...er and data cables as shown in Connecting iQue Power and Data Cables 2 Prepare fluids and fill the bottles and troughs as shown in iQue Fluids 3 Place a tube of filtered distilled water on the detecto...

Page 92: ...86 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C Special Topics First Time Startup Checklist...

Page 93: ...DO NOT select the Shutdown Detector option 2 Perform a Long Clean as shown in Standard Cleaning Procedures to fully decontaminate the system 3 Click Power Off Detector in the Controller Console window...

Page 94: ...88 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C Special Topics Long Term Shutdown Checklist...

Page 95: ...p and also any time that the instrument needs to be physically moved to a new location The iQue Uninterruptible Power Supply Each iQue comes with an Uninterruptible Power Supply UPS This UPS is requir...

Page 96: ...tery backed up sockets on the UPS this is not required for safety but having the computer on battery backup can prevent work interruption in the case of a power outage CAUTION The iQue s UPS is requir...

Page 97: ...91 iQue Hardware Manual Part Number 10554 Rev C Special Topics Connecting iQue Power and Data Cables 110V Power and Data Connection Diagram 220V Power and Data Connection Diagram...

Page 98: ...92 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C Special Topics Connecting iQue Power and Data Cables...

Page 99: ...the need to package and ship an iQue arises contact IntelliCyt Tech Support Referenced Procedures Standard Cleaning Procedures Daily Shutdown First Time Startup Checklist Shutting Down before a Move...

Page 100: ...hick piece of foam 4 Be gentle when setting the system down 5 Lift by gripping the base tray using the recessed cups near each of the four feet 6 Do NOT lift by the bottle drawer or plate rail extensi...

Page 101: ...brated in the field Before performing deck position calibration contact IntelliCyt Tech Support for assistance to be sure that the problem is not a result of another issue such as an improperly calibr...

Page 102: ...With a Gen 1 iQue the user must lower the probe manually in order to determine the proper Z position In all other respects deck position calibration is the same for Gen 1 and Gen 2 iQue instruments L...

Page 103: ...up the Calibrate iQue screen which contains links for the various deck locations that can be calibrated Note that for a Gen 1 iQue there is no Testing Point and the first menu item only says Calibrat...

Page 104: ...il it just touches the shaker deck surface as shown Probe just touching deck Be sure that the probe itself is not sliding up in the guide If the probe has been pushed upward the deck height calibratio...

Page 105: ...ht button The contact sensing probe will descent and measure the deck height CAUTION Proper test point calibration is essential for proper iQue functioning Do not attempt this procedure without assist...

Page 106: ...y in higher density formats Note that during Well Plate calibration depth is not recorded Sampling depth is a function of deck height and Plate Model See Managing Plate Models above Calibrate Fluid St...

Page 107: ...iQue Hardware Manual Part Number 10554 Rev C Special Topics Re Calibrating Deck Positions 3 Move the probe to a position over each one of the test points in turn until all the wizard steps are comple...

Page 108: ...102 THIS PAGE INTENTIONALLY LEFT BLANK iQue Hardware Manual Part Number 10554 Rev C Special Topics Re Calibrating Deck Positions...

Page 109: ...IntelliCyt Tech Support IntelliCyt Corporation 9620 San Mateo NE Albuquerque NM 87113 USA www intellicyt com Telephone 505 345 9075 opt 2 E mail support intellicyt com...

Page 110: ......

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