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Contents

vii

Chapter 5: Troubleshooting

Instrument Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Fluidics Cart Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

101

Loader Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

102

Appendix A: Technical Overview

105

Flow Cytometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

106

Fluidics System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

106

Optics System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

109

Light Scatter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

110

Fluorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

111

Optical Filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

112

Detectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

114

Detector Arrays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

114

Spillover . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

117

Electronics System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

119

Pulses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

120

Pulse Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

121

Appendix B: Optics Configurations

125

4-2 Optics Configuration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

126

4-2-2 Optics Configuration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

130

5-3 Optics Configuration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

134

Summary of Contents for FACSCanto II

Page 1: ...2347 Brazil Tel 55 11 5185 9995 Fax 55 11 5185 9895 Europe Tel 32 53 720211 Fax 32 53 720452 Mexico Tel 52 55 5999 8296 Fax 52 55 5999 8288 Japan Nippon Becton Dickinson Company Ltd Tel 0120 8555 90 A...

Page 2: ...any means whatsoever except as otherwise permitted by law BD FACSCanto clinical software Becton Dickinson and Company This software is the property of Becton Dickinson and Company Each sale of a stor...

Page 3: ...h this unit to ensure compliance with the Class A FCC limits This Class A digital apparatus meets all requirements of the Canadian Interference Causing Equipment Regulations Cet appareil num rique de...

Page 4: ......

Page 5: ...t 26 Computer Workstation 29 BD FACS Loader Optional 29 Barcode Reader Optional 30 System Requirements 31 Chapter 2 BD FACS Loader Option 33 Loader Components 34 Using the Loader 36 Opening and Closin...

Page 6: ...luidics System Long Clean 59 Replacing the Air Filter 60 Replacing Fluidics Filters 63 Unscheduled Maintenance 66 Changing a Cubitainer 68 Removing an Air Lock 72 Cleaning External Surfaces 74 Removin...

Page 7: ...verview 105 Flow Cytometry 106 Fluidics System 106 Optics System 109 Light Scatter 110 Fluorescence 111 Optical Filters 112 Detectors 114 Detector Arrays 114 Spillover 117 Electronics System 119 Pulse...

Page 8: ...r Replacement Parts 141 Bar Code Reader Parts 141 Consumables 142 Instrument Setup 142 Reagents 142 Labware 143 Appendix D Technical Specifications 145 Cytometer Specifications 146 Environment 147 Per...

Page 9: ...ating algorithms while BD FACSDiva software is non application specific You can use either software to perform instrument quality control Each software package has its own reference manual provided on...

Page 10: ...remains unchanged Symbol Meaning CAUTION hazard or unsafe practice that could result in material damage data loss minor or severe injury or death Electrical danger Laser radiation Biological risk Tabl...

Page 11: ...tact your local BD Biosciences technical support representative or supplier When contacting BD Biosciences have the following information available product name part number and serial number software...

Page 12: ...xii BD FACSCanto II Flow Cytometer Reference Manual...

Page 13: ...r cooled lasers and provides its own air pressure and vacuum for the fluidics and waste You can prepare samples on the BD FACS Sample Prep Assistant II and import the sample prep worklist For further...

Page 14: ...Cytometer Reference Manual Intended Use The BD FACSCanto II system is intended for use as an In Vitro Diagnostic device for identification and enumeration of lymphocyte subsets in human cells in suspe...

Page 15: ...ined fluidics cart and the BD FACSCanto II workstation System options include an automated sample loader and a barcode reader For a description of system components see Flow Cytometer on page 16 Fluid...

Page 16: ...tem an optics subsystem and an electronics subsystem For a more in depth discussion of fluidics optics electronics and flow cytometry see the Technical Overview on page 105 Fluidics The fluidics syste...

Page 17: ...tor arm movable waste aspiration port aspirator arm bar metal bar used to push aspirator arm away from SIT during manual loading tube guide helps guide tube onto the SIT Loader operation only tube eje...

Page 18: ...software In BD FACSDiva software SIT cleaning between tubes is automatic unless you disable it by deselecting the SIT Flush checkbox on the Acquisition Dashboard Do not leave a tube of distilled wate...

Page 19: ...ur fluorescence signals from a blue laser source and two fluorescence signals from a red laser source The 4 2 2 configuration adds a violet laser and can collect two fluorescence signals from the viol...

Page 20: ...e laser light to a focusing lens The lens directs the laser light onto the sample stream within the flow cell Figure 1 3 See also Figure A 3 on page 109 Figure 1 3 Optical pathway figure shows violet...

Page 21: ...onsist of photomultiplier tubes PMTs arranged in one octagon and one trigon as shown in Figure 1 4 The octagon contains five PMTs and detects light from the 488 nm blue laser One PMT in the octagon co...

Page 22: ...e next highest wavelengths and reflect lower wavelengths and so on around the array A bandpass filter or additional longpass mirror in front of each PMT further screens unwanted light In addition to t...

Page 23: ...ight from interfering with fluorescence signal Table 1 2 Octagon and trigon optical filters 4 2 configuration Detector Array Laser PMT Position LP Mirror BP Filter or LP Mirror Intended Dye Octagon 48...

Page 24: ...board the fluidics controllers and the acquisition indicator lights This section describes only the user accessible power panel and the acquisition indicator lights For more information see Electroni...

Page 25: ...s Lights are activated only when the system is acquiring and only the indicators corresponding to currently active parameters will blink The acquisition indicator lights can be switched off The on off...

Page 26: ...ng fluids to the cytometer and collects system waste products The cart supplies the required air pressure and vacuum which eliminates the need for an external source although the cart can be hooked up...

Page 27: ...ed with the system Do not substitute other containers Figure 1 8 Fluidics cart containers and ports Each solution has its own non interchangeable fluid port and level sensor connection Fluid level ala...

Page 28: ...ave the auxiliary air supply switch off unless the cart has been attached to an in house air supply by BD Biosciences service personnel see Table 1 3 for details Leave the cart circuit breaker on at a...

Page 29: ...e daily startup procedure Computer Workstation The workstation consists of a computer compatible with Microsoft Windows XP Professional operating system a flat screen monitor and an optional printer B...

Page 30: ...arcode standards including Codabar Code 128 Code 39 with checksum and PDF417 It reads information from the BD FACS 7 color setup beads label into BD FACSCanto clinical software and also reads coded pa...

Page 31: ...through BD Biosciences Compatible Tubes 12 x 75 mm polystyrene test tubes BD Falcon tubes 12 x 75 mm BD Trucount tubes BD FACS 7 color setup bead tubes Bulk Fluids BD FACSFlow solution BD FACSClean s...

Page 32: ...ACSCanto II Flow Cytometer Reference Manual Fluids Required for External Cleaning BD FACSClean solution Deionized DI water Setup Beads BD FACS 7 color setup beads for use with BD FACSCanto clinical so...

Page 33: ...33 2 BD FACS Loader Option This chapter contains the following information Loader Components on page 34 Using the Loader on page 36 Maintaining the Loader on page 42...

Page 34: ...ription tube guide with sensor guides tubes during lifting to ensure a proper seal sensor indicates if tube guide is in position for Loader operation alignment guide pin aligns the carousel and keeps...

Page 35: ...the carousel slides out for easy access center spindle the axis around which the carousel spins insert the carousel onto the Loader here drive system the motor that rotates the carousel not visible fr...

Page 36: ...ning the Loader Doors To open the front door pull the top of its handle toward you away from the cytometer to disengage the latch and then slide the door to the left To open the right door use its han...

Page 37: ...he Loader 1 Verify that the cytometer is set up for automatic loading The tube guide should be forward and the aspirator arm bar should be horizontal and aimed to the front toward the carousel Figure...

Page 38: ...rtex the sample tubes and place them uncapped in the carousel s according to the worklist For accurate results match the tubes to those listed on the printed worklist BD FACSCanto clinical software or...

Page 39: ...op of the carousel Press down firmly to seat the carousel Figure 2 4 Seating the carousel in the Loader 5 Close the Loader drawer completely and close the Loader cover doors The Loader scans and posit...

Page 40: ...age 41 Changing to Manual Loading 1 Open the Loader cover doors 2 Pull out the drawer 3 Remove any carousel in the Loader 4 In manual mode you do not need the tube guide arm Move it all the way back a...

Page 41: ...ally Changing to Automatic Loading 1 If the aspirator arm bar is not already in the forward position rotate it to a horizontal position and aimed to the front toward the carousel Handle all biological...

Page 42: ...ill Containment Area The top of the Loader has a depressed area intended to contain spills that may occur during operation Check this area for fluids and clean as described in the following section Al...

Page 43: ...ter power and unplug the AC power cord from the wall socket 2 Wipe all accessible surfaces with BD FACSClean solution Ensure that the optical sensors are cleaned Saline deposits can develop on the sen...

Page 44: ...44 BD FACSCanto II Flow Cytometer Reference Manual...

Page 45: ...45 3 Barcode Reader Option This chapter contains the following information Installing and Using the Barcode Reader on page 46 Cleaning the Barcode Reader on page 48 Barcode Symbologies on page 49...

Page 46: ...software 2D barcode symbology reading coded patient information from sample tube labels into a worklist 1D barcode symbology NOTICE The barcode reader supplied with the BD FACS Sample Prep Assistant I...

Page 47: ...rmation dialog from the setup values label supplied with the setup bead kit NOT from the lot number on the setup bead tubes Patient information is scanned from sample labels into the worklist ID field...

Page 48: ...from the barcode while continuing to hold the trigger Tip To obtain a reading keep the bar code reader aimed at the center of the label Do NOT sweep across the label 7 Compare software field values wi...

Page 49: ...ormation transfer including sample ID assignments This can result in a mismatch of sample IDs and sample results By default the barcode reader has checksums enabled We recommend you do not disable che...

Page 50: ...ACSCanto clinical software 2D barcode symbology is required to read all target values with one scan For information on installing and using the barcode reader with the BD FACSCanto II system see Insta...

Page 51: ...em requires basic preventive maintenance to preserve instrument reliability This section explains the procedures you should follow to keep your instrument in good condition Scheduled Maintenance on pa...

Page 52: ...D FACS shutdown solution Refer to BD FACSCanto II Instructions for Use Daily Empty the waste See Emptying the Waste Container on page 54 Daily and as needed Wipe down SIT and aspirator arm Prevents sa...

Page 53: ...59 Monthly and before service calls Replace the air filter Ensures proper instrument performance See Replacing the Air Filter on page 60 Every 6 months Replace fluid filters Keeps fluids free of part...

Page 54: ...them can transmit potentially fatal disease To prevent exposure to biohazardous agents expose waste container contents to bleach 10 of total volume before disposal Dispose of waste in accordance with...

Page 55: ...p from the container place the assembly on the bench label side up The waste tank can become pressurized when the cytometer is running Always disconnect the tank from the fluidics cart before you empt...

Page 56: ...the trap Replace it with a new cap Write the date on the cap label 7 Screw the cap assembly onto the tank and hand tighten until it is fully closed 8 Re attach the sensor and fluid lines To prevent ta...

Page 57: ...Prime fluid lines before purging the filters as noted in the procedure NOTICE The cytometer must be on system pressurized during this procedure Materials paper towels proper protective equipment Proce...

Page 58: ...it until fluid seeps out Fluid should seep from the open valve within 30 seconds If no fluid appears make sure the corresponding cubitainer is not empty or detached from the cart If the cubitainer con...

Page 59: ...solution approximately 275 mL BD FACS shutdown solution approximately 1 100 mL Procedure 1 Check all fluid levels Empty the waste if needed 2 Select Cytometer Instrument Cleaning Modes Long Clean A c...

Page 60: ...utdown solution from the interior reservoir and fluid lines and replaces it with BD FACSFlow solution Tip If setup fails after a long clean repeat Fluidics Startup until setup passes Replacing the Air...

Page 61: ...e 61 Materials replacement filter see Other Replacement Parts on page 141 for ordering Procedure 1 Turn off the cytometer power 2 Open the side door Press the black button a handle will pop out Twist...

Page 62: ...long the upper edge of the filter to release the old filter Dispose of the old filter It cannot be reused 4 Install a new filter Ensure that the arrows etched on edges of the new filter point in towar...

Page 63: ...door Push door closed until the door latches Push the handle in to latch the handle Replacing Fluidics Filters Change the fluid filters when you see increased debris in FSC vs SSC plots or every 6 mo...

Page 64: ...ordering paper towels proper protective equipment felt tip pen Procedure 1 Place a few paper towels beneath the filter to collect drips 2 Remove the filter by pressing the tabs on each quick disconnec...

Page 65: ...Wait until fluid seeps out Fluid should seep from the open valve within 30 seconds If no fluid appears ensure the corresponding cubitainer is not empty or detached from the cart If the cubitainer con...

Page 66: ...Lock on page 72 When fluidics are not functioning properly flow cell will not fill or there is backflush into sample tube Clean external surfaces Keeps surfaces free from salt buildup after exposure...

Page 67: ...Resetting the Cytometer Circuit Breaker on page 85 After the circuit breaker is tripped Reattach Ethernet and RS232 cables See Reconnecting the Ethernet and RS232 Cables on page 86 When a cable is di...

Page 68: ...uration Each cubitainer and the waste tank has its own color coded port Connect containers to the following ports Container Port Label Port Color waste Waste A orange BD FACSFlow solution BD FACSFlow...

Page 69: ...ring events 2 Detach the sensor and fluid line from the cart Pull the sensor straight out Press the metal clip on the fluid line 3 Unscrew the cap on the cubitainer You could damage the sensor line if...

Page 70: ...7 Reattach the sensor line and fluid line to the cart To attach the sensor line gently rotate until the connection aligns and then push To attach the fluid line push the coupling into the port until i...

Page 71: ...ubitainer or detach the fluidics lines for other maintenance 1 Choose Cytometer Instrument Cleaning Modes Prime After Tank Refill 2 Select the checkboxes for the cubitainers you changed click OK 3 Cli...

Page 72: ...any of the fluidics filters Materials paper towels bypass tubing Procedure 1 Place a few paper towels beneath the air locked filter to collect drips 2 Remove the filter by pressing the metal tabs Figu...

Page 73: ...ar a click 4 Choose Cytometer Instrument Cleaning Modes Prime After Tank Refill 5 Select the checkbox that corresponds to the filter you have bypassed 6 Click OK 7 When the prime finishes remove the b...

Page 74: ...en exposed to sheath fluid NOTICE Over time saline deposits may develop on the interior surface of the SIT Bal seal retainer causing an incomplete SIT flush between samples If this occurs you might ne...

Page 75: ...rfaces to prevent corrosion 4 Wipe all exposed surfaces with a clean dry cloth All instrument surfaces that come in contact with biological specimens can transmit potentially fatal disease Use univers...

Page 76: ...ubbles repeat the process Cleaning the Flow Cell Use the Clean Flow Cell command to run a tube of BD FACSClean solution through the SIT and flow cell After the procedure is complete the cleaning fluid...

Page 77: ...ayed during the cleaning 4 After the completion message displays wait 5 minutes Waiting allows BD FACSClean solution to dissolve deposits in the flow cell cuvette 5 Click OK 6 Remove the tube from the...

Page 78: ...ubble filter Poor CVs might indicate a need to purge the bubble filter 1 Start the bubble filter purge In BD FACSCanto clinical software select Cytometer Cleaning Modes Bubble Filter Purge In BD FACSD...

Page 79: ...through 4 in Decontaminating the Fluidics System Long Clean on page 59 2 Shut down the software and turn off the power to the cytometer 3 Wipe down the SIT and the aspirator arm with DI water Replaci...

Page 80: ...II Flow Cytometer Reference Manual Figure 4 5 Bal seal and retainer Materials proper protective equipment Bal seal removal tool o replacement Bal seal DI water retainer SIT Bal seal use this end to r...

Page 81: ...ter 2 If you have a Loader installed Open the Loader cover doors Pull out the drawer If needed remove the carousel Move the tube guide to manual position See Changing to Manual Loading on page 40 3 Un...

Page 82: ...rence Manual 4 Lower the retainer down the SIT and let it rest on the aspirator arm 5 Access the Bal seal at the notch on the right side of the SIT assembly Unseat the Bal seal by using the Bal seal r...

Page 83: ...slide down the SIT and rest on top of the retainer 7 Center the Bal seal on the retainer With one hand support the retainer and Bal seal and with the other hand move the aspirator arm to the left Lowe...

Page 84: ...old the new Bal seal spring side up see figure and slide it up the SIT Use both hands to gently push it into its seated position Tip If the Bal seal is not quite in its seated position replacing the r...

Page 85: ...shown 11 Test the installation by manually loading a tube onto the SIT and running fluid 12 If you are using a Loader ensure that the system is ready for automatic loading See Changing to Automatic L...

Page 86: ...use the following diagram to reconnect them Both cables connect to ports on the PC workstation As the make and model of the workstation might vary refer to the documentation that came with your system...

Page 87: ...rt labeled i Fluid Out See Table 4 2 on page 88 for port functions Table 4 1 Correspondence of cytometer ports to fluidics cart ports Port or Button on Cytometer Port on Fluidics Cart a System Power b...

Page 88: ...cart b Power Out Connects to fluidics cart c Air In 70 5 psi d Waste A Waste out aspirated e Communications Data port f Sheath B BD FACSFlow solution port g Waste A Waste out non aspirated h On Off A...

Page 89: ...page 141 for ordering proper protective equipment If you are changing the sensor on the waste tank use proper precaution and wear suitable protective clothing eyewear and gloves All biological specime...

Page 90: ...ference Manual Procedure 1 Ensure that the cytometer is not acquiring events 2 Detach the sensor and fluid line from the cart 3 Unscrew the cap on the cubitainer 4 Remove the level sensor assembly and...

Page 91: ...you replaced any level sensors other than the waste sensor prime the affected fluidics lines Important Continue with the procedure in Priming Fluidics Lines on page 71 To ensure that the appropriate...

Page 92: ...all screwdriver Removing Old Fuses 1 Shut down the cytometer and turn off the instrument power 2 Unplug the power cord from the wall outlet and the fluidics cart Removing the plug allows easier access...

Page 93: ...out the fuse drawer until you can grip it 5 Remove the fuse drawer and remove and dispose of the old fuses Tip Note the positions of the old fuses before you remove them Duplicate these positions wit...

Page 94: ...voltage is right side up 8 Slide the drawer back into the instrument until it snaps into place 9 Close the fuse access door 10 Reconnect the power cord to the fluidics cart 11 Plug the cytometer power...

Page 95: ...Manual the BD FACSDiva Software Reference Manual or the BD FACSCanto II Instructions for Use If you need additional assistance contact BD Biosciences Refer to our website bdbiosciences com for up to d...

Page 96: ...tton located on the left side of the cytometer to turn the system off and on No pressure in the plenum Check air supply components see Figure 4 6 on page 87 Check air supply connections Check air supp...

Page 97: ...scribed in Removing an Air Lock on page 72 Fluid backfill into sample tube Cracked tube Transfer sample to new tube Make sure you are using appropriate tubes See System Requirements on page 31 Bal sea...

Page 98: ...tometer BD FACSDiva software running at same time as BD FACSCanto clinical software Quit the software you are not using or put the cytometer in standby Cytometer Instrument Standby Cytometer power is...

Page 99: ...lled or worn Reinstall or replace the Bal seal See Replacing the Bal Seal on page 79 Liquid leakage around cytometer base Interior valve failure 1 Turn off the cytometer power 2 Clean up the liquid us...

Page 100: ...2 Clean up the liquid using proper precautions 3 Call BD Biosciences Waste line to fluidics cart disconnected 1 Turn off the cytometer power 2 Clean up the liquid using proper precautions 3 Check that...

Page 101: ...off the cytometer power 2 Clean up the liquid 3 Check and close all bleeder valves See Purging the Fluidics Filters on page 57 Broken fluid line Contact BD Biosciences Fluidics cart will not power on...

Page 102: ...lure Contact BD Biosciences Tube missed or missing tube messages in software Salt crystal buildup on optical sensors Clean the tube optical sensor outer sensor as described in Cleaning External Surfac...

Page 103: ...e alignment guide pin engages with the alignment hole and press down See step 4 on page 39 If the problem persists contact BD Biosciences for assistance Aspirator arm bar vertical Rotate the bar horiz...

Page 104: ...104 BD FACSCanto II Flow Cytometer Reference Manual...

Page 105: ...5 Appendix A Technical Overview This appendix provides more information about these topics Flow Cytometry on page 106 Fluidics System on page 106 Optics System on page 109 Electronics System on page 1...

Page 106: ...Fluidics System A fluidics system in a flow cytometer moves particles in fluid through a flow cell past a laser beam and then into a waste tank Sheath Cubitainer to Flow Cell On the BD FACSCanto II fl...

Page 107: ...ssure in BD FACSCanto clinical software by displaying the Status window In BD FACSDiva software select Instrument Instrument Status Report sheath tank sheath filter interior reservoir plenum bubble fi...

Page 108: ...and sheath fluid stream can be used to vary the diameter of the sample stream also known as the sample core Increasing the sample pressure increases the core diameter and therefore the flow rate Figu...

Page 109: ...nce emission has a very brief duration only a few microseconds This brief flash of light is collected filtered and then converted into an electrical signal by the detectors Figure A 3 Light pathway th...

Page 110: ...am is called forward scatter FSC light that scatters roughly perpendicular to the laser beam is called side scatter SSC FSC and SSC intensities indicate physical properties of cells FSC indicates rela...

Page 111: ...rgy photon than the excitation wavelength Some fluorescent compounds emit at a much longer wavelength than their excitation wavelength PerCP absorbs blue light 488 nm and emits red light 675 nm other...

Page 112: ...ffect of an optical filter on incident photons Two kinds of filters are used on the BD FACSCanto II flow cytometer default configuration longpass LP bandpass BP Longpass Filter In general LP filters m...

Page 113: ...lters are typically designated by two numbers The first number indicates the center wavelength and the second refers to the width of the band of light that is passed For example a 500 50 BP filter tra...

Page 114: ...n BD FACSDiva software you can use the Instrument Configuration dialog to specify the fluorochrome that will be measured at each PMT detector You can also add additional parameters to your configurati...

Page 115: ...on 4 2 configuration Behind the LP mirror a 780 60 BP filter admits light from 750 to 810 nm and substantially blocks other wavelengths The light that finally reaches PMT A will be from dyes such as P...

Page 116: ...elength will pass through to another LP filter 670 nm that further blocks shorter wavelength light Light from dyes such as PerCP Cy5 5 pass through this filter The beam continues around the array with...

Page 117: ...C into PE detector For example FITC appears primarily in the FITC detector but some of its fluorescence spills over into the PE detector PE appears primarily in the PE detector but some of its fluores...

Page 118: ...fluoresce in the yellow 575 nm range so there is no way to isolate the emission from each fluorochrome optically Instead fluorescence compensation moves the FITC population out of the PE positive are...

Page 119: ...of digital electronics requires a basic understanding of the bit Computers and digital circuits use bits binary numbers consisting of ones and zeros to pass information A 4 bit number has 4 binary di...

Page 120: ...ser beam At this point both the beam intensity and signal intensity are low The pulse reaches a maximum intensity or height when the particle reaches the middle of the beam where the beam and signal i...

Page 121: ...ion times per second slicing it into 16 384 levels and assigning a measurement to each time sample PMT preamp ADC digital delay PMT preamp ADC digital delay PMT preamp ADC digital delay PMT preamp ADC...

Page 122: ...r further processing During this process when a pulse exceeds the user assigned threshold its height and area are simultaneously calculated by the FPGA Figure A 17 Pulse measurements An FPGA calculate...

Page 123: ...2 144 levels In order to normalize the height to the area measurement the electronics multiples the height by 16 16 384 x 16 262 144 To more finely adjust the area to match the height scale the DSP ap...

Page 124: ...124 BD FACSCanto II Flow Cytometer Reference Manual...

Page 125: ...Configurations This appendix describes the components for each of the BD FACSCanto II optics configurations 4 2 Optics Configuration on page 126 4 2 2 Optics Configuration on page 130 5 3 Optics Conf...

Page 126: ...al power and cooling requirements Collection Optics The BD FACSCanto II 4 2 configuration collection optics include one octagon and one trigon detector array The octagon contains five PMTs and detects...

Page 127: ...Intended Dye Octagon 488 nm blue laser A 735 780 60 PE Cy7 B 655 670 PerCP Cy5 5 or PerCP C 610 blank optical holder D 556 585 42 PE E 502 530 30 FITC F blank optical holder 488 10 and pinhole SSC G b...

Page 128: ...meter Reference Manual Figure B 1 4 2 configuration octagon detector array 488 nm blue laser 5 3 0 3 0 585 42 5 5 6 L P P E F I T C 5 0 2 L P 670 LP 655 LP 7 8 0 6 0 7 3 5 L P 6 1 0 L P P E C y 7 PerC...

Page 129: ...Appendix B Optics Configurations 129 Figure B 2 4 2 configuration trigon detector array 6 8 5 L P 780 60 735 LP 660 20 APC Cy7 APC 633 nm red laser...

Page 130: ...ic focusing lens as shown in Figure 1 3 on page 20 The 4 2 2 configuration uses low powered air cooled and solid state lasers that do not have special power and cooling requirements Collection Optics...

Page 131: ...BP Filter or LP Mirror Intended Dye Octagon 488 nm blue laser A 735 780 60 PE Cy7 B 655 670 PerCP Cy5 5 or PerCP C 610 blank optical holder D 556 585 42 PE E 502 530 30 FITC F blank optical holder 48...

Page 132: ...eter Reference Manual Figure B 3 4 2 2 configuration octagon detector array 488 nm blue laser 5 3 0 3 0 585 42 5 5 6 L P P E F I T C 5 0 2 L P 670 LP 655 LP 7 8 0 6 0 7 3 5 L P 6 1 0 L P P E C y 7 Per...

Page 133: ...B Optics Configurations 133 Figure B 4 4 2 2 configuration trigon detector arrays 450 50 510 50 502 LP AmCyan Pa ci fic B lu e 405 nm violet laser 6 8 5 L P 780 60 735 LP 660 20 APC Cy7 APC 633 nm re...

Page 134: ...figuration uses low powered air cooled and solid state lasers that do not have special power and cooling requirements Collection Optics The BD FACSCanto II 5 3 configuration collection optics include...

Page 135: ...ntended Dye Octagon 488 nm blue laser A 735 780 60 PE Cy7 B 655 670 PerCP Cy5 5 or PerCP C 610 616 23 PE Texas Red D 556 585 42 PE E 502 530 30 FITC F blank optical holder 488 10 and pinhole SSC G bla...

Page 136: ...Manual Figure B 5 5 3 configuration octagon detector array 5 3 0 3 0 585 42 5 5 6 L P P E F I T C 5 0 2 L P 670 LP 655 LP 7 8 0 6 0 7 3 5 L P 6 1 0 L P P E C y 7 PerCP Cy5 5 PerCP SS C 488 nm blue la...

Page 137: ...Appendix B Optics Configurations 137 Figure B 6 5 3 configuration trigon detector array 712 21 6 8 5 L P 780 60 735 LP 660 20 APC Cy7 APC A le xa Fl uo r 70 0 633 nm red laser...

Page 138: ...138 BD FACSCanto II Flow Cytometer Reference Manual...

Page 139: ...placement Parts To order supplies and options contact your local BD Biosciences representative For updated information and part numbers refer to our website bdbiosciences com Instrument Supplies on pa...

Page 140: ...r US power 15A 5 15P 320 C13 337219 2 5 m cordset for Australian power 10A C13 335696 2 5 m cordset for European power 10A C13 335697 2 5 m cordset for UK power 10A C13 R A 335698 Filter bypass assemb...

Page 141: ...979 Waste sensor probe 2 level 338978 Auxiliary sensor probe 1 level 343835 Air filter side door 336303 Bal seal retainer 640116 Carousel kit carousels 1 4 332727 Carousel kit carousels 5 8 332728 Car...

Page 142: ...eath fluid BD Biosciences 877 232 8995 340398 US and Latin America 342003 other countries BD FACSClean solution BD Biosciences 340345 BD FACS shutdown solution BD Biosciences 334224 Monoclonal antibod...

Page 143: ...143 Labware Item Supplier Catalog No 5 mL polystyrene test tubes 12 x 75 mm BD Falcon tubes uncapped 125 per bag capped 125 per bag capped 25 per bag with cell strainer cap 25 per bag BD Biosciences...

Page 144: ...144 BD FACSCanto II Flow Cytometer Reference Manual...

Page 145: ...Specifications Cytometer Specifications on page 146 Fluidics Cart Specifications on page 150 BD FACS Loader Specifications on page 151 For barcode reader specifications refer to the information suppl...

Page 146: ...in between unit and other objects or wall to permit proper air flow and access to the main power button and circuit breaker Right side 30 cm 11 8 in between unit and other objects or wall to permit pr...

Page 147: ...ve humidity 20 80 noncondensing Noise level 62 dBA Facilities No special room requirements Fluorescence threshold sensitivities FITC 100 MESF PE 50 MESF Forward and side scatter sensitivity Platelets...

Page 148: ...rical aperture NA 1 2 Fluorescence detection 6 to 8 photomultiplier tube detectors Wavelength ranges detected from 488 nm laser 750 810 nm PE Cy7 670 735 nm PerCP Cy5 5 610 637 nm PE Texas Red optiona...

Page 149: ...60 L min High 120 L min Sample acquisition rate 10 000 events sec with 10 abort rate 8 parameters Recommended maximum particle size 50 m Workstation resolution 262 144 channel resolution Data acquisi...

Page 150: ...between air vent and other objects or wall to permit proper air flow Fluidics cart door air vent 20 cm 7 9 in between door and other objects or wall to permit proper air flow Weight 51 kg 112 lb fluid...

Page 151: ...er carousels numbers 1 16 Tube compatibility Carousel Accommodates up to 40 uncapped 12 x 75 mm tubes BD Falcon polystyrene test tubes BD Trucount tubes BD FACS 7 color setup bead tubes Thickness of a...

Page 152: ...152 BD FACSCanto II Flow Cytometer Reference Manual...

Page 153: ...l 80 Bal seal retainer 79 saline deposits 74 bandpass BP filters 113 barcode reader 30 cleaning 48 focal lengths 48 installing 46 symbologies 49 BD FACS Loader 29 BD FACS Sample Prep Assistant II 13 B...

Page 154: ...configuration optics 4 2 19 126 4 2 2 19 130 5 3 19 134 connecting power cords 24 connection fluid port 27 level sensor connection 27 container waste 27 cubitainer BD FACS shutdown solution 27 BD FACS...

Page 155: ...tion 27 required 31 fluidics about 106 level sensors replacing 89 long clean 59 priming 71 system components 16 fluidics cart components 26 controls 28 dimensions 150 fuses replacing 92 maintenance 54...

Page 156: ...Uniphase HeNe Air Cooled 20 126 130 134 Point Source iFLEX 130 power 24 red 19 safety interlock 20 solid state 20 specifications 147 troubleshooting 99 violet 19 leaks cytometer 99 fluidics cart 101 S...

Page 157: ...alignment 34 plenum 16 18 about 106 PMT 114 power button 16 24 instrument 24 laser 24 pressure sheath fluid 107 priming fluid lines 71 prisms 20 beam shaping 20 pulse 120 area 122 height 122 purging...

Page 158: ...barcode 49 system requirements 31 T technical assistance xi specifications 151 trap condensation 29 trigon 114 optics 21 23 troubleshooting 102 tube eject cylinder 17 guide 34 37 about 17 sensor 17 w...

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Summary of Contents for FACSCanto II

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BD FACSCanto II, Reference Manual, Page: 7 / 158

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Summary of Contents for FACSCanto II

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