BDAccuri C6 user manual
Contents
1. ccccccecceceeeeeeeeeeeeeeeseeeeeseeeeseeeeseaeeeseeeesseeeees 5 3 2 1 Setting UP the Computer cece ceccccceeeeceeeeeceeeeseeeeeseeeesseeessaeeeseeesseeesaaees 5 3 2 2 Changing the Workstation Computer Settings for Windows 7 0 0000 6 3 2 3 Viewing c6 and c6t File Extensions in WiINdOWS 7 cccccseceeeeseeeneeeneeenes 6 3 3 Setting Up the BD Accuri C6 Flow Cytometer cccccceeeeceeeeeeeeeeseeeeeseeeeseeeeeneeeess 7 3 3 1 Setting Up the Reagent Bottles and Fluidics ec cceeecceeeeeeeeeeeeeeeeesaeeees 8 3 3 2 Connecting the Cables cccccccccscccccseceeeeeeeseeeeseeceeseeeeseeeessaeeesensesseeesaaees 9 3 4 Installing Software 0 00 ec eceecccceececeeeeceeeeeceeeeeseeeeseueessaeeeseusessaueeseaeessaeeeseueesseeesees 10 3 4 1 Installing BD Accuri C6 Softwa re cccccccccecccseeeeceeeeseeeeeseeeesaeeeeseeeesaaeeesees 10 3 5 Starting Up the BD Accuri C6 Flow Cytometer cccccceeeeeeeeeseeeeeeeeeeseeeeseeeesees 10 3 6 Turning Off the BD Accuri C6 Flow Cytometer cccccecseeeeeeeeeeeeeeeeeeseeeesaeeeesees 11 4 MAINTENANOE xct nctsceccecacencadetecnesdecodedscsestnddeuisanesuedeuadsus eesabneocs entcanetetsenqposaseceststedouedsene 13 4 1 Automated Preventive Maintenance Notification ccccccccseccsseeeeeeeeeneeeneeeneeenees 13 A 212 01016 Mg loa gee ee ee ee ee ae ne eee er eee ee eee 14 43 Cleaning We FIOW Cel asrini2. Change the setting using the slider under the Resolution field s drop down menu if necessary c Select Apply then select OK and close the Personalization window 3 2 3 Viewing c6 and c6t File Extensions in Windows 7 To view hidden file extensions which distinguish BD Accuri C6 Software files c6 from BD Accuri C6 Software template files c6t 1 OT oa Se wY Click on the Windows Start button and select Control Panel from the pop up menu Select Tools from the menu bar Click on View tab Click Show hidden files and folders Deselect Hide extensions for known file types 7820018 01 Rev 2 6 BD Accuri C6 Flow Cytometer Instrument Manual Select Apply then OK to save changes and close dialog box 3 3 Setting Up the BD Accuri C6 Flow Cytometer 1 2 3 Remove the cytometer and other components from the shipping boxes With the front of the cytometer facing toward you lift the top cover Verify that the in line sheath filter is installed If it is not installed see section 4 11 for instructions on replacing the filter Figure 3 2 In line Sheath Filter Verify that there are no disconnected electronic cables on the optics assembly Verify that the interference filters are firmly seated Figure 3 3 The standard interference filters are labeled as follows e FL1 533 30 e FL2 585 40 e FL8 6 70LP e FL4 675 25 Interference filter
3. Figure 3 3 Interference Filter Never run the cytometer without interference filters installed Running the cytometer without the filters may damage the instrument 7820018 Rev 2 7 BD Accuri C6 Flow Cytometer Instrument Manual 6 Gently close the cytometer cover Make sure the cover is securely positioned If the cover is not closed securely the cytometer will not function Place the fluidics bottle tray in a convenient location near the perimeter of the cytometer A Do not place the fluidics bottle tray on top of the cytometer 3 3 1 Setting Up the Reagent Bottles and Fluidics Fill the sheath bottle with sheath fluid Dilute the decontamination concentrate PN 653154 and cleaning concentrate PN 653157 solutions as instructed and fill decontamination and cleaner bottles respectively Ay Close the bottles securely Do not over tighten 3 Add approximately 100 mL of 0 5 NaOCl to the Waste bottle Place the boitles in the fluidics bottle tray Fluidics harness Figure 3 4 Fluidics harness Slide the red tab on the black end of the fluidics harness to the unlock position and attach the black end of the fluidics harness to the back of the cytometer Slide the red tab to the lock position 7 Attach the other end of the fluidics harness to the appropriate bottles e Red line waste bottle e Blue line sheath bottle e Yellow line decontamination bottle e Green line cleaner bottle 7820018 01
4. Ordening IPOMMANOM eacesacssencs ccustcanuscsenpacapbsagccuetcapscupivapszosesapseehtvaseoundsaeecebesepeseaaetse 27 7820018 Rev 2 i BD Accuri C6 Flow Cytometer Instrument Manual FIGURES AND TABLES Figure 1 1 The BD Accuri C6 Flow Cytometer System cccccceeccceeeeeceeeeceeeeseeeeeseesessaeeesens 1 Table 1 3 BD Accuri C6 Flow Cytometer Specifications cccccsecceceeeeeseeeeeeeeesseeeeeeeeseeeeees 2 Table 2 1 Approved Reagents xc civncesscsucctece ss davela a a a aa 3 Figure 3 1 BD Accuri C6 Flow Cytometer Optics and Fluidics Top View Lid Open 5 Figure 3 2 In line Sheath Finer ancienne We heeded a a a ia 7 Figure 3 3 Mern eronee Pile saa cere reed need ne ET TE 7 Figure 3 4 Fl dies MANNS S serine a a a a 8 Figure 3 5 Proper Attachment of Fluidics Lines cece cccecceeeceeeeceeeeaeeeseeeeeeesaeeeseeeeseeesaeess 9 Figur S 60 Cabe CONNECCION S kecakan aa 9 Figure 3 7 SIP and Sample Stage cccccscccceeccccecescsecenceceecsecesseeccceececsecescenccoseeesceeeseeeeess 10 Figure 3 8 Improper Shut down Me SSaQEC cccccccsececeeeceececueeceeeceucecueecueessueeseeeeaeeessusenaess 11 Table 4 1 BD Accuri C6 Flow Cytometer Replacement Parts Reagents List 0cceeeee 13 Figure 4 1 Automated Preventive Maintenance Notification WiINdOW cccceeeeseeeeeeeeeeees 14 Figure 4 2 Waste Line Quick DiISCONNECL ccccccceeseceeceeeeceecceeee
5. every two months Replace both pieces of tubing at the same time Tubing comes in contact with biological samples and therefore should be considered hazardous Wear gloves and appropriate safety gear during this procedure There are two peristaltic pumps a sheath pump and a waste pump 1 Turn off and unplug the cytometer 2 Lift the cytometer lid 3 Disconnect the luer locks from the peristaltic pumps by unscrewing them Blue tubing is connected to the sheath pump and red tubing to the waste pump Sheath pump Waste pump Figure 4 4 Peristaltic Pumps and Luer Locks 4 Squeeze the grip marks on the pump element retainer clip to remove the clip Figure 4 5 Removing Peristaltic Pump Retainer Clip 7820018 Rev 2 19 BD Accuri C6 Flow Cytometer Instrument Manual D 9 Pull the luer lock connectors outward and slide the luer lock fittings off the pump head Figure 4 6 Sliding Luer Lock Fittings Off of Pump Head Remove the peristaltic pump tubing and discard according to standard laboratory protocols and regulations Replace with new peristaltic pump tubing PN 653146 by sliding the luer lock fittings on the pump head Snap the pump element retainer clip into place Reconnect the luer lock fittings on the outside of the pumps Ensure the blue and red tubes are not crimped or over tightened A Ensure that the luer lock fittings are connected to the correct tubing elements Genily close the cytometer
6. For a thorough cleaning of the flow cell perform an extended flow cell clean During extended flow cell cleaning the flow cell fills completely with cleaning solution from the sample tube on the SIP This cycle automatically shuts down the cytometer with cleaning reagent in the flow cell allowing the flow cell to soak 1 Place a tube with at least 500 uL of Extended Flow Cell Clean Solution PN 653159 on the SIP Ay Never run the Extended Clean of Flow Cell cycle without a tube containing at least 500 uL of fluid Select nstrument gt Extended clean of flow cell 3 After the cytometer is automatically shut down allow the cytometer to rest for at least 30 minutes up to overnight for a more thorough cleaning 4 Replace the tube of Extended Flow Cell Clean Solution with a tube of 2 mL 0 22 um filtered DI water 5 Restart the cytometer The cytometer performs a longer fluidics startup cycle and the Software displays the message Extra startup time needed due to cleaning or improper shutdown This longer cycle purges cleaning reagent from the flow cell Run 0 22 um filtered DI water for 10 minutes on fast speed 7 Operate the cytometer as usual Decontamination Fluid Cycle The BD Accuri C6 automatically runs the decontamination fluid cycle when the system is shut down normally The decontamination fluid cycle lasts about 15 minutes To manually run the decontamination fluid cycle 1 Place a tube with 2mL of 0 22 um filte
7. Rev 2 BD Accuri C6 Flow Cytometer Instrument Manual Figure 3 5 Proper Attachment of Fluidics Lines 3 3 2 Connecting the Cables 1 Plug the cord from the power supply into the back of the cytometer Power jack USB port BD CSampler serial cable used with BD CSampler only Figure 3 6 Cable Connections 2 Attach the black power cord to the power supply and plug into a standard outlet Ay Only operate this instrument from a supply source that incorporates a third wire protective grounding conductor A surge suppressor or power strip is recommended 3 Connect the USB cable to the back of the cytometer A Do not plug the USB cable into the computer yet A To prevent putting pressure on the fluidics lines and USB cable connection do not place the cytometer against a wall or another object 7820018 Rev 2 9 BD Accuri C6 Flow Cytometer Instrument Manual 3 4 3 9 10 Installing Software This section provides details on installing BD Accuri C6 Software and the BD Accuri C6 flow cytometer driver In most cases BD Accuri C6 Software will be pre installed If software is installed skip to section 3 5 3 4 1 Installing BD Accuri C6 Software 1 2 3 Ensure the USB cable is not connected between the computer and cytometer Start up the computer and monitor Insert the provided BD Accuri C6 Software CD In the CD directory double click on BD Accuri C6 Software Installer exe Follow the instructio
8. changes to the C6 workstation computer in order to achieve the best possible work environment and to avoid unexpected software computer issues that could potentially result in a loss of data 3 2 2 Changing the Workstation Computer Settings for Windows 7 Skip this section if your computer has BD Accuri C6 Software preinstalled 1 Go to Start circular icon located in the lower left hand corner select Control Panel Select Appearance and Personalization 3 Turn off the Sleep Settings a From the column on the left select System and Security then Power Options b Select Change when the computer sleeps from the left hand margin alternatively select change plan settings from the main window c From the Edit Plan Settings window change the Put the computer to sleep setting to Never using the drop down function we also suggest setting the Turn off the display setting to Never d Select Save Changes Then close the control panel window Optimize Monitor Display Settings From Start Control Panel Appearance and Personalization a Select Display select Adjust Resolution from the column on the left Alternatively select Adjust screen resolution directly from the Display section of the main Appearance and Personalization window Then select highlight Monitor 1 b Ensure the Resolution is set to the maximum setting or to at least 1152 x 864
9. if you are having difficulty performing this procedure due to instrument malfunction or you need further guidance on the process Limited Warranty Limited Warranty period is one 1 year from the earlier of i first operation of the Products by Buyer or ii ten 10 days after shipment of the Products to Buyer This Limited Warranty covers software and firmware and defects in material and workmanship under normal use and does not apply to ordinary wear and tear For more detailed information refer to the warranty information at www AccuriCytometers com Extended Warranties Extended warranties are available Visit www AccuriCytometers com for information Technical Support For Technical Support contact In the USA Accuri Cytometers Inc 173 Parkland Plaza Ann Arbor MI 48103 USA Phone 1 734 994 8000 Fax 1 734 994 8002 Email TechSupport AccuriCytometers com 7820018 Rev 2 25 BD Accuri C6 Flow Cytometer Instrument Manual e Europe Middle East and Africa BD Biosciences Erembodegem Dorp 86 9320 Erembodegem Belgium Phone 32 53 720 882 Customer Support bd accuri europe bd com Technical Support techsupport europe bd com e Canada BD Biosciences 2100 Derry Rd West Suite 100 Mississauga Ontario Canada L5N 0B3 Email canada bd com Direct Phone Numbers Austria 0810 101 807 Belgium 078 166 050 Canada 1 800 268 5430 Denmark 8025 0622 Finland 0800 915582 France 0811 290 069 Germany 01
10. the beads are out of performance specification perform a decontamination fluid cycle see section 4 4 and run the beads again lf they are still out of specification call Accuri Technical Support The event rate drops off while running a sample but the cytometer is not clogged 1 Place a tube of cleaning solution on the SIP and run for five minutes 2 Run 0 22 um filtered DI water for one minute on Fast fluidics speed 3 Check to see if the event rate is restored The cytometer pumps are running normally but BD Accuri C6 Software is not acquiring data 1 Check the levels in the sheath and waste bottles 2 Check to see if any of the external fluidic lines are crimped 3 Runa backflush to see if the SIP is clogged see section 4 2 4 Make sure the peristaltic pump tubing and retainer clips are properly attached see section 4 12 5 Restart BD Accuri C6 Software The cytometer pumps are running continually 1 Check for a large air bubble in the in line sheath filter If so replace the filter see section 4 11 2 There could be a clog in the line Run backflush or unclog 3 The peristaltic pump tubing may need to be replaced see section 4 12 For additional technical support view the frequently asked questions FAQs at www AccuriCytometers com customer_support fags or contact Accuri Technical Support if a problem persists Cleaning and Decontamination Standard Procedure This procedure should be perf
11. to repair the instrument Any evidence of fluid coming from or near a red or clear line should be considered a biological hazard Do not attempt to clean up the fluid without gloves and other safety attire Dispose of all cleaning materials as if biologically contaminated Replacing the In line Sheath Filter Replace the in line sheath filter PN 653148 every two months If the cytometer is used daily for more than a few hours replace the filter monthly Wear appropriate safety attire such as protective gloves and eyewear when performing this procedure 1 Turn off and unplug the cytometer Genily lift the cytometer cover Remove the plastic storage compartment Twist the luer locks on the in line sheath filter to disconnect the locks oO oe a Discard the filter according to standard laboratory protocols and regulations In line sheath filter Figure 4 3 In line Sheath Filter 6 Install a new in line sheath filter PN 653148 This filter has a male and female end to ensure that it can only be installed in the correct orientation 7 Replace the storage compartment 7820018 01 Rev 2 BD Accuri C6 Flow Cytometer Instrument Manual 8 Gently close the cytometer cover 9 Place a sample tube with 2mL of 0 22 um filtered DI water on the SIP 10 Plug in and turn on the cytometer 11 Purge any air from the instrument see section 4 6 4 12 Replacing the Peristaltic Pump Tubing Replace the pump tubing PN 653146
12. 80 100 1732 Ireland 1850 930 396 ltaly 840 999 926 Netherlands 0900 0400 142 Norway 800 17 382 Poland 00800 121 4744 26 7820018 01 Rev 2 5 6 BD Accuri C6 Flow Cytometer Instrument Manual Ordering Information Visit www AccuriCytometers com or www BDBiosciences com for a complete list of parts and reagents Regional Offices bdbiosciences com offices Asia Pacific Singapore Tel 65 6861 0633 Fax 65 6860 1590 China Tel 86 21 3210 4610 Fax 86 21 5292 5191 India Tel 91 124 2383566 Fax 91 124 2383224 25 26 Canada BD Biosciences Toll Free 888 259 0187 Tel 905 542 8028 Fax 888 229 9918 bdbiosciences com ca canada bd com Australia New Zealand Australia Toll Free 1800 656 100 Tel 61 2 8875 7000 Fax 61 2 8875 7200 bd anz bd com New Zealand Toll Free 088 572 468 Tel 64 9 574 2468 Fax 64 9 571 2469 bd anz bd com Japan Nippon Becton Dickinson Toll Free 0120 8555 90 Tel 81 24 593 5405 Fax 81 24 593 5761 Europe BD Biosciences Tel 32 2 400 98 95 Fax 32 2 401 70 94 bdbiosciences com eu help biosciences europe bd com United States BD Biosciences Toll Free 877 232 8995 Fax 800 325 9637 bdbiosciences com answers bd com Microsoft Office and Windows 7are trademarks of Microsoft Corporation 2012 For Research Use only Not for diagnostic or therapeutic procedures Class 1 Laser Product BD Accuri BD Accuri C6 Software BD Accuri CSampler Software BD CSampler and Scien
13. BD Accuri C6 Flow Cytometer Instrument Manual Science is hard Flow cytometry should be easy BD Accuri C6 Flow Cytometer Instrument Manual TABLE OF CONTENTS T INTRODUC ON erreneren e a e eE ee E Senet ocshren shee Rese bsupbeapecenGsapsousteagect 1 De ADU TAS MNA ea E E E ede aes 1 1 2 Delivery and Inspection scccsccctwadadiasindedsasendadnameuasinateedednmmuanenbenindnatenas niieadeinaerandnatendd 1 1 3 BD Accuri C6 Flow Cytometer Specifications cccccccseececeeeeeeeeeeseeeeseeeeeseeesaeeeees 2 2 SAFETY PRECAUTIONS scpcacecacecsucccensatecocsdsncceuadeth Sncpccuscouedetednepdsassocete ecedn cusseancakedecpesuceee 3 2 1 Safety and Limitations Manual cccccccccecccseeeeceeeseeeeeseeeeseeeeeseeeeseeeessaeeeseeesaeeeess 3 2 2 Operational Considerations ccccecccseececeececeececeeeeseeeeeseeeeseueeeseueeseueesseeeseeessneeess 3 2 2 1 Ambient Temperature and Humidity cccccseccseceeeeeseeeseeeseeeseeeseeeneeeneeenes 3 222 PAETOS 3 2 2 3 Approved Reagents cccccsecccseeeceeeeeeeeeeeeeseeeseueeseueeseeeseueeseeesseeeseeeeseeesaeeess 3 2 2 4 Material Safety Data Sheets MSDS ccc ecccceeceeseeeeeeeeeeseeeeseeesseeeesaeees 4 3 SETTING UP THE BD ACCURI C6 FLOW CYTOMETER cccccccseeeeeeeeeeeeseeeeeeeeeeeeens 5 3 1 BD Accuri C6 Flow Cytometer Layout cccccccceececeeeeseeeeceeeeeseeeeseesesseeeeseeeesseeees 5 3 2 Configuring the Computer Settings
14. ce is hard Flow cytometry should be easy are trademarks of Becton Dickinson and Company 2012 7820018 Rev 2 2
15. cover 10 Place a sample tube with sheath on the SIP 11 Plug in and turn on the cytometer 12 Purge air from the cytometer see section 4 6 20 7820018 01 Rev 2 BD Accuri C6 Flow Cytometer Instrument Manual 5 TROUBLESHOOTING AND TECHNICAL SUPPORT This section provides troubleshooting and technical support information 5 1 Troubleshooting This section contains basic troubleshooting guidelines Follow the procedures listed below If problems persist after performing troubleshooting contact Accuri Technical Support The cytometer and or computer does not power on 1 Make sure the power supplies and cords are plugged into an appropriate outlet 2 Check the outlet to make sure it is functioning properly The acquisition controls on BD Accuri C6 Software do not show a green light and remain a gray color after the cytometer is powered on 1 Make sure the USB cable is properly attached to the cytometer and computer It may take several seconds for the port to recognize the cytometer Switch the USB cable to a different port on the computer Make sure the BD Accuri C6 flow cytometer driver was properly installed on the computer by doing the following in order Double click the BD Accuri C6 Software icon on your computer desktop to open BD Accuri C6 Software See the BD Accuri C6 Software User s Guide or Quick Start Guide Collection for instructions on using BD Accuri C6 Software Connect the USB cable from
16. e Solution for treating 10 L Sheath Fluid 10 vials Cleaning Concentrate Solution makes 1 L BD Accuri C6 Flow Cytometer Fluid Kit contains concentrate for making 1 L each of Cleaning and Decontamination Solutions Bacteriostatic Concentrate for treating 10 L of Sheath and 8 mL of Extended Flow Cell Clean Extended Flow Cell Clean Solution BD Accuri C6 Flow Cytometer Starter Kit Contains concentrate to make 1 L each of Cleaning and Decontamination Solutions Bacteriostatic Concentrate to treat 10 L of Sheath 8 mL of Extended Flow Cell Clean and 4 mL of both 6 and 8 Peak Validation Beads Visit www accuricytometers com for a complete list of available parts and reagents Automated Preventive Maintenance Notification An automated notification and reminder system is contained in the BD Accuri C6 Software to alert the user to perform the recommended preventive maintenance as described in this manual This includes removal and replacement of the peristaltic pump tubing in line sheath filter and the fluidic bottle filters 7820018 Rev 2 13 BD Accuri C6 Flow Cytometer Instrument Manual 4 2 4 3 14 BD Accuri C6 Software will trigger the notification based on the volume of sheath run through the system or on the date since maintenance was last performed BD Accuri C6 Software will display this notification based on whichever trigger is reached first The notification appears as a pop up window that allows the user to choose b
17. eeeaeeeceesseeeceessaeeeeeesaaaeeess 17 Figure 4 3 In line Sheath Filter scvosdecsiccescaccccyssnactvacdasteses ances EEE AEE E Ri 18 Figure 4 4 Peristaltic Pumps and Luer LOCKS cccceecceeeceeeceeeceeeeeueeeeeeeeeeeueeauesaueeaeeeaeeeaaes 19 Figure 4 5 Removing Peristaltic Pump Retainer Clip ccecccseecseeeceeeseeeseeeeseeeeseeesaeeeseees 19 Figure 4 6 Sliding Luer Lock Fittings Off of Pump Head ccccceececeeeeseeeteeeeceeeeseeeseeeeneees 20 Figure 5 1 Browse For Folder Dialog BOX c ccccceeccseeeeeeeeceeeeseeeseeeseeeesaeesaueeseeeseeesaueesaees 22 Figure 5 2 Update Driver Software Cytometer Second Dialog BOX cccccseeeeseeeeeeeeees 22 Figure 5 3 Windows Security WINKOW cccccccccseeeeseeeeeeeeeeseeeesseeeeseeeesaeeeeseeeesaaeeeseeeseeeeas 23 Figure 5 4 Update Driver Software Cytometer Successfully updated your driver software 23 Figure 5 5 The C6 Software Status Message Box C6 is connected and ready 066 23 ii 7820018 01 Rev 2 1 1 1 1 2 BD Accuri C6 Flow Cytometer Instrument Manual INTRODUCTION The BD Accuri C6 flow cytometer offers an entirely new vision for the role of flow cytometry in life science research Designed from the ground up to be compact and easy to use the BD Accuri C6 flow cytometer fits in any lab and is readily accessible to all researchers experts and novices alike Figure 1 1 The BD Accuri C6 Fl
18. emperature of the cytometer is between 15 C and 30 C 59 F and 86 F Operating humidity is lt 80 relative humidity 2 2 2 Area of Use The cytometer is designed to fit on a laboratory bench top Keep a perimeter of at least six 6 inches 15 cm around the instrument and computer for proper ventilation and to protect the instrument and computer from liquid spills Take special care while handling fluids around the cytometer Ensure that uncontained fluids do not enter the instrument or computer Clean up spills immediately Never place anything on top of the cytometer including sample tubes and racks Turn off the instrument and unplug the power cord before manually cleaning the cytometer 2 2 3 Approved Reagents Only approved fluids should be used in the operation or cleaning of this instrument See the following table for a list of approved reagents Table 2 1 Approved Reagents Sheath fluid 0 22 um filtered deionized DI water with or without bacteriostatic concentrate solution PN 653156 Cleaning concentrate solution PN 653157 diluted according to package instructions Decontamination concentrate solution PN 653154 diluted according to package instructions Extended flow cell clean Extended flow cell cleaning solution PN 653159 A Do not use household bleach as a decontamination solution It contains fluorescent whitening agents that may interfere with fluorescence detection BD Accuri C6 Flow Cytometer I
19. etween Remind me later or Maintenance is complete Selecting Maintenance is complete will re start both triggers Selecting Remind me later will set the notification to remind the user in 5 days Maintenance Required Avoid high background counts and blocked valves Inform the maintenance supervisor to replace the Following 1 Reagent bottle filters 2 In line sheath Filter 3 Tubing in the peristaltic pumps C Maintenance is complete e Figure 4 1 Automated Preventive Maintenance Notification Window Cleaning the SIP Run the backflush cycle as needed to clean the SIP and remove clogs at the base of the SIP 1 Place a blotter or empty sample tube under the SIP to catch dripping fluid 2 Doone of the following e Click on the Backflush button in the Collect tab e Select Instrument gt Run Backflush Cycle Cleaning the Flow Cell Clean the flow cell as a part of regular maintenance or to correct performance issues of the cytometer 4 3 1 Running the Unclog Cycle The unclog cycle purges the flow cell of debris 1 Remove the sample tube from the SIP and place a blotter or empty sample tube under the SIP to catch dripping fluid 2 Doone of the following 7820018 01 Rev 2 4 4 4 5 BD Accuri C6 Flow Cytometer Instrument Manual e Click on the Unclog button in the Collect tab e Select Instrument gt Run Unclog Cycle 4 3 2 Running an Extended Clean of the Flow Cell
20. ght turns green and BD Accuri C6 Software displays the message C6 is connected and ready run 0 22 um filtered DI water for at least 15 minutes before processing samples Validate cytometer performance See the BD Accuri C6 Software User s Guide for details on instrument validation using the validation beads provided with the cytometer 3 6 Turning Off the BD Accuri C6 Flow Cytometer 1 a i i Place a tube with 2 mL of decontamination solution on the SIP Select an empty data well in the Collect Tab of BD Accuri C6 Software Set the time limit for two minutes and the fluidics speed to fast Click on the RUN button Once the run is finished remove the tube of decontamination solution from the SIP Place a tube with 2 mL of 0 22 um filtered DI water on the SIP and advance to any empty data well Set the time limit for two minutes and fluidics to Fast Click on the RUN button 9 When the run is finished leave the tube on the SIP Press the power button The shutdown cycle runs for approximately 15 minutes then the cytometer automatically powers off Ad beresin the power button for five seconds or longer bypasses the auto fluidic shutdown cycle If the cytometer is shut down in this manner BD Accur C6 Software displays the following message during the next start up Extra startup time needed due to cleaning or improper shutdown Figure 3 8 Improper Shut down Message When this occurs the cytometer
21. his driver software anyway Only install driver software obtained from your manufacturer s website or disc Unsigned software from other sources may harm your computer or steal information v See details Figure 5 3 Windows Security Window e Windows will start the installation of the driver for the C6 cytometer Once completed you have successfully installed the driver software When prompted click the Close button Ss 8 Update Driver Software Cytometer Windows has successfully updated your driver software Windows has finished installing the driver software for this device Cytometer T Figure 5 4 Update Driver Software Cytometer Successfully updated your driver software e With the C6 driver installed and the USB cable properly connected BD Accuri C6 Software will indicate that it recognizes the C6 by displaying a green traffic light and the message C6 is connected and ready C6 is connected and ready Figure 5 5 The C6 Software Status Message Box C6 is connected and ready 7820018 Rev 2 23 BD Accuri C6 Flow Cytometer Instrument Manual 5 2 A A 24 Data does not look as expected 1 Pause and restart the sample 2 Run a backflush or unclog see sections 4 2 and 4 3 1 3 Run the validation beads PN 653144 or 653145 see the BD Accuri C6 Software User Guide for details If the beads look acceptable there may be a problem with the sample If
22. na RR EE n ETER 14 4 3 1 Running the Unclog Cycle ccccccccssecccseecceeeeceueeceeseeceueessueeeceusessueessaas 14 4 3 2 Running an Extended Clean of the Flow Cell s nsnennnennnennnennnenennnrenreesnenne 15 4 4 Decontamination Fluid Cycle 2 0 0 cccccccccccsececseeeeceeeesseeeeseeesseeeeseeessaeeeseeeesseeeesees 15 45 Cleaning Fluid Cycle seisseen aaa aas 15 4 6 Purging Air from the BD Accuri C6 Flow Cytomete r cccsecceceeeeeeeeeeseeeeaeeeeeees 16 47 Emptying th Waste CONN ON dsssseisarrei gense aAA EEE 16 AO FINNS Fud BONG so eeeee e eee eee 17 4 9 Replacing the Fluidic Bottle Filters cc eccccccseeeeeeeeeeeeeeeeecaeeeeeeeeeeeseeeaeeeneeenaes 17 4 10 Inspecting the Fluidics LINES cee ccecceeeceeeeeeeeeeeeeeeeeeeeeeeaeeeaueeeeeeaeeeneeeaeeeneeenaes 18 4 11 Replacing the In line Sheath Filter cc ccccccccsececseeeeseeesseeeesesesseeeeseesesseeesees 18 4 12 Replacing the Peristaltic PUMP Tubing ccccccseccseeeeeeeeeeeeeeeaeeeneeeneeeneeeneeeneeenees 19 5 TROUBLESHOOTING AND TECHNICAL SUPPORT ccccccceeeeceeeeeeseeeeesaeeeeeseeeeeeas 21 Oat Troubleshooting essersi Eaa AERAR AREER EE RENERE 21 5 2 Cleaning and Decontamination Standard Procedure cccccseeecseeeeeeeeneeeseeeens 24 DS LEN A er E E E E EEEE R 25 oa ENOO G I aE aea E E 25 3 83 Rel 81 612 Ee 0 0 g a ao enn arr een Se Der set tert Oar Senne eee ene ene eee 25 56
23. ng This is normal Click on the Run button again wait for 30 seconds and click on the Pause button to stop the pumps Repeat step 5 two more times to purge bubbles from the system Emptying the Waste Container Empty the waste bottle daily or when prompted by BD Accuri C6 Software to prevent spillover A Biological samples are potentially dangerous and or life threatening Adhere to proper handling procedures for samples and reagents at all times Wear appropriate laboratory attire such as protective gloves and eyewear during this procedure 1 Disconnect the quick connect waste line from the top of the waste bottle 7820018 01 Rev 2 BD Accuri C6 Flow Cytometer Instrument Manual Waste line quick disconnect Figure 4 2 Waste Line Quick Disconnect Remove the bottle lid Dispose of waste according to local regulations Add approximately 100 mL of 0 5 NaOCl to the Waste bottle Replace the lid D pN Reattach the waste line to the top of the bottle Filling the Fluid Bottles Before turning on the cytometer visually check all the bottles and fill the sheath decontamination and cleaning solution bottles as needed NOTE The cytometer is a non pressurized system If necessary any of the bottles can be opened while the cytometer is powered on NOTE BD Accuri C6 Software displays a message when the fluid bottles need attention or if a bottle level sensor is improperly connected Replacing the Fluidic Bo
24. ns in the installation wizard Starting Up the BD Accuri C6 Flow Cytometer 1 Check fluid levels in all bottles ensuring that the Waste is empty and the Sheath Cleaner and Decontamination bottles are full Gently push the sample stage back remove and discard the SIP cover and place a tube containing at least 2 mL of 0 22 um filtered DI water on the SIP SIP Collar SIP Sample Stage Figure 3 7 SIP and Sample Stage NOTE The sample stage can accommodate several types of tubes including any brand of 12x75 mm tube and most microcentrifuge tubes While holding the sample tube in place pull the sample stage forward to support the tube The stage clicks into place A The sample tube does not form a seal at the top of the SIP While inserting the SIP in the tube be careful not to bend or catch the SIP on the outside of the tube 7820018 01 Rev 2 4 BD Accuri C6 Flow Cytometer Instrument Manual Firmly press the power button on the front of the unit to turn on the cytometer During start up the BD Accuri C6 Software Traffic Light turns yellow and the cytometer pumps start to run When powered on the cytometer automatically flushes the fluidics lines with sheath This process takes approximately five minutes A Do not open the lid of the cytometer during the startup process Opening the lid interrupts the laser warm up and extends the time before data can be acquired When the BD Accuri C6 Software Traffic Li
25. nstrument Manual 2 2 4 Material Safety Data Sheets MSDS To obtain an MSDS visit www AccuriCytometers com or contact e Inthe USA Email TechSupport AccuriCytometers com Call Accuri Technical Support at 1 734 994 8000 e Outside the USA Visit www AccuriCytometers com for country specific contact information 4 7820018 01 Rev 2 3 3 1 3 2 7820018 Rev 2 BD Accuri C6 Flow Cytometer Instrument Manual SETTING UP THE BD ACCURI C6 FLOW CYTOMETER Perform all of the procedures in this chapter before running any experimental samples BD Accuri C6 Flow Cytometer Layout The cytometer is composed of fluidics optics and electronics subsystems that work together to analyze cell populations The following figures show the major components of the fluidics and optics subsystems Sheath pump Waste pump Blue laser Red laser Optical assembly In line sheath filter Figure 3 1 BD Accuri C6 Flow Cytometer Optics and Fluidics Top View Lid Open Configuring the Computer Settings 3 2 1 Setting up the Computer Set up the computer monitor keyboard and mouse according to manufacturer s instructions Computers come preinstalled with Windows 7 Professional Contact Accuri Technical Support for assistance with other operating systems BD Accuri C6 Flow Cytometer Instrument Manual A Install the software on the computer before connecting the USB cable to the cytometer NOTE We recommend making the following
26. ormed when preparing the system for storage or shipping it for service Contact Accuri Technical Support for more information on shipping Visit www AccuriCytometers com for information on service contracts Take appropriate safety measures such as wearing gloves and proper laboratory attire while performing this procedure 1 Turnonthe Cytometer and open the BD Accuri C6 Software 7820018 01 Rev 2 5 3 5 4 5 9 BD Accuri C6 Flow Cytometer Instrument Manual Place a sample tube containing 3 mls of diluted Decontamination Fluid PN 653154 on the SIP and run the instrument for 5 minutes on FAST Place a sample tube containing 3 mls of 0 22 um filtered DI water on the SIP and run the instrument for 2 minutes on FAST From the Instrument menu in BD Accuri C6 Software activate the manual decontamination fluid cycle by selecting Run Decontamination Fluid Cycle 5 When the Decontamination Fluid Cycle is complete turn off the BD Accuri C6 Remove tube of 0 22 um filtered DI water from SIP Disconnect the fluidic harness from the C6 and the fluid bottles empty the four fluid bottles and wipe down the outside of the instrument with 70 ethanol Pack the C6 fluid bottles and tray fluidic harness and power supply in the Pelican Case A When disconnecting the fluidic harness from the cytometer always disconnect from the bottle end of the harness first to prevent fluid leaks NOTE Contact Accuri Technical Support
27. ow Cytometer System The BD Accuri C6 flow cytometer system includes BD Accuri C6 Software for controlling the instrument generating statistics and analyzing results About This Manual This manual provides information on cytometer requirements and specifications setup and maintenance It also includes common troubleshooting help and service information e For information on using BD Accuri C6 Software see the BD Accuri C6 Software User Guide e For information on installing and using the BD CSampler see the BD Accuri CSampler Software User Guide Delivery and Inspection Carefully inspect all cartons upon receipt of the cytometer If there are any signs of mishandling or damage file a claim with the carrier immediately If the shipment is separately insured file a claim with the insurer ITEMS NOT PROVIDED e Surge suppressor or uninterruptible power supply UPS A model rated at a minimum of 1000 VA is recommended for use with the cytometer computer and monitor e Microsoft Office or equivalent e Anti virus software This system is intended for use as a stand alone station not as part of a network 7820018 01 Rev 2 1 BD Accuri C6 Flow Cytometer Instrument Manual 1 3 BD Accuri C6 Flow Cytometer Specifications Table 1 3 BD Accuri C6 Flow Cytometer Specifications 488 nm rated at 20 000 hr life 640 nm rated at 20 000 hr life 20 mW 488 nm Solid State Blue Laser Side 90 degrees 13 4 colors s
28. red DI water on the SIP 2 Select Instrument gt Run decontamination fluid cycle Cleaning Fluid Cycle The cleaning fluid cycle pulls cleaner fluid from the cleaner tank and runs it through the fluidic lines of the fluidics system After filling the system with cleaner fluid the cleaning fluid cycle purges the cytometer with fresh sheath fluid and performs a backflush This cycle takes about five minutes 1 Place a tube of cleaning solution PN 653157 on the SIP 2 Select Instrument gt Run cleaning fluid cycle 7820018 Rev 2 15 BD Accuri C6 Flow Cytometer Instrument Manual 4 6 16 Purging Air from the BD Accuri C6 Flow Cytometer Purge air from the cytometer if any of the following is true The cytometer is being set up for the first time The cytometer has not been used for more than two days A sample has been run dry or the cytometer has been running without sheath in the sheath bottle To purge air from the cytometer 1 Place a tube containing sheath on the SIP NOTE If performing an air purge at any time other than initial setup skip steps 2 3 Select Instrument gt Run decontamination fluid cycle 3 Wait approximately 15 minutes for the decontamination fluid cycle to complete Set the run time in the software control panel to five minutes and click on the Run button Allow the cytometer to run until it stops automatically NOTE The peristaltic pumps will make a rhythmic sound while runni
29. s Cytometer Driver Date Driver Version Digital Signer Search for driver software in this location Driver Details _ Update Driver dete Driver V Include subfolders Roll Back Driver Dss Browse for driver software on your comp gt Let me pick from a list of device drii This list will show installed driver software co ec software in the same category as the device Figure 5 1 Browse For Folder Dialog Box e Click the Next button Update Driver Software Cytometer Browse for driver software on your computer Search for driver software in this location C Program Files BD Accur BD Accuri C6 Software libraries Include subfolders gt Let me pick from a list of device drivers on my computer This list will show installed driver software compatible with the device and all driver software in the same category as the device Figure 5 2 Update Driver Software Cytometer Second Dialog Box e When the Windows Security dialog box shown below opens click on nstall this driver software anyway 22 7820018 01 Rev 2 BD Accuri C6 Flow Cytometer Instrument Manual J Windows Security amp Windows can t verify the publisher of this driver software gt Don t install this driver software You should check your manufacturer s website for updated driver software for your device gt Install t
30. takes additional time to recover and return to the green light ready state 7820018 Rev 2 11 BD Accuri C6 Flow Cytometer Instrument Manual NOTE It is not necessary to exit BD Accuri C6 Software or turn off the computer before turning off the cytometer 12 7820018 01 Rev 2 4 4 1 BD Accuri C6 Flow Cytometer Instrument Manual MAINTENANCE Changing the in line sheath filter fluidic bottle filters and peristaltic pump tubing every two months is recommended Heavy use of the cytometer may warrant more frequent changes The following table lists the parts and reagents that are required for regular maintenance Table 4 1 BD Accuri C6 Flow Cytometer Replacement Parts Reagents List 653146 653147 653148 653149 653150 653154 653155 653156 653157 653158 653159 653160 Peristaltic Pump Tubing Set of 2 Sheath bottle filter 1 large and Decon Cleaner Bottle Filters 2 small In line Sheath Filters 1 ea BD Accuri C6 Flow Cytometer 1 year Maintenance Kit contains 12 Peristaltic Pump Tubing 6 Sheath Bottle Filter 12 Decontamination Cleaner Bottle Filters and 6 In line Sheath Filters BD Accuri C6 Flow Cytometer 2 Month Maintenance Kit contains 2 Peristaltic Pump Tubing 1 Sheath Bottle Filter 2 Decon Cleaner Bottle Filters and 1 In line Sheath Filter Decontamination Concentrate Solution makes 200 mL Decontamination Concentrate Solution makes 1L Bacteriostatic Concentrat
31. tandard optical filters e FL1 533 30 nm e g FITC GFP Emission Detection e FL2 585 40 nm e g PE PI e FL8 gt 670nm e g PerCP PerCP Cy 5 5 PE Cy7 e FL4 675 25 nm e g APC Minimum Detectable Particle 0 5 um Size Minimum Sample Volume 300 uL in 12 x 75 mm tube Pre Set Flow Rates and Core Slow 14 uL min 10 um core Sizes Medium 35 uL min 16 um core Fast 66 uL min 22 um core 0 22 um filtered deionized DI water 1 million events per well MESF monocytes and granulocytes 2 0 05 for chick erythrocyte nuclei CEN lt 3 CV for CEN 100 240 VAC 50 60 Hz Instrument Size 11 H x 14 75 W x 16 5 D 27 9 x 37 5 x 41 9 cm 11 H x 21 5 W x 16 5 D 27 9 x 54 6 x 41 9 cm 30 Ibs 13 6 kg Fluid bottle capacity 2 L sheath fluid 2 L waste 250 mL cleaner 250 mL decontamination Signal Processing 24 bit datapath Computer Interface USB 2 0 MESF values determined using Thermo Scientific Cyto Cal Multifluor Plus Violet Beads 2 7820018 01 Rev 2 2 2 1 2 2 7820018 Rev 2 BD Accuri C6 Flow Cytometer Instrument Manual SAFETY PRECAUTIONS Safety and Limitations Manual Refer to the BD Accuri C6 Safety and Limitations Manual PN 7820091 for complete safety information Operational Considerations When setting up the cytometer note the details in the following sections to maximize the instrument s performance 2 2 1 Ambient Temperature and Humidity Operating t
32. the C6 to one of the computer USB ports CAUTION Do not plug the USB cable for the cytometer into any USB ports on the computer monitor NOTE Windows will attempt to auto install the BD Accuri C6 driver The attempt will end with a message stating Device driver was not successfully installed This is as expected 7820018 Rev 2 After the C6 Device driver fails to auto install Click the Start button round button with the Windows logo on the lower left of the screen Select Devices and Printers You should see an icon labeled Cytometer that has a yellow symbol with an exclamation point in it Right click on the Cytometer icon and select Properties Once the properties window opens select the Hardware tab With the hardware tab open click the Properties button Click the Change Settings button When the settings window opens click the Update Driver button When the driver update window opens select Browse my computer for driver software 21 BD Accuri C6 Flow Cytometer Instrument Manual e Navigate to the location of the driver software and specify the following path C Program Files BD Accuri BD Accuri C6 Software libraries Then click on the OK button to enter the selected path into the Browse window Add a device Add a printer 4 Unspecified 1 J Cytometer Properties General Hardware i Cytometer Cytometer Properties General Driver Details Device Function
33. ttle Filters Each of the sheath cleaner and decontamination bottles contain a disk filter Replace these filters PN 653147 every two months Wear appropriate safety attire such as protective gloves and eyewear when performing this procedure 1 Disconnect the quick connect lines from the top of each bottle 2 Carefully remove the lid from each bottle 3 Disconnect the filter luer lock at the end of the fluidic tubing Discard the filter according to standard laboratory protocols and regulations 4 Replace the filter with the appropriate part e Sheath bottle large disk filter e Cleaner and decontamination bottles small disk filter 5 Reassemble the bottles and reconnect the quick connect lines 7820018 Rev 2 17 BD Accuri C6 Flow Cytometer Instrument Manual 4 10 A 4 11 18 6 Place a tube with 2 mL of 0 22 um filtered DI water on the SIP and run for one minute Inspecting the Fluidics Lines Check the cytometer periodically to ensure there are no fluid leaks or crimped lines 1 Turn off the cytometer 2 Open the top cover 3 Visually inspect for fluid leaks by looking for small pools of liquid near any of the visible luer lock connections 4 Visually inspect for dried residue or slight discoloration of the metal fluidics tray plate making sure to look under the glovebox and under the cytometer 5 If any evidence of a leak is seen contact Accuri Technical Support immediately Do not attempt
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