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Cytometer Configurations

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CantoII - WIMR 

Baseline: 

Baseline reports:  

Cytometer Baseline Report_cantowimr.pdf

CS&T bead lot: 93308

Laser (nm)

Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen2.5x rSDen

405

50





1



V450_50B

Pacific Blue, Cascade Blue, AF405, V450, BV421, 

450/50

NA


23.5

58.75



V510_50A

CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO

510/50

502LP


22

55

488 

20





2



SSC

SSC

488/10

NA


NA

 NA



B530_30E

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, SYBR Green

530/30

502LP


22.1

55.25



B585_42D

PE,PI,DsRed,DiA

585/42

556LP


19.8

49.5



B670_LPB

PerCP,PI, PerCP-Cy5.5,PerCP-Cy5,PE-Cy5,7AAD, BB700

670LP

655LP


21.7

54.25



B780_60A

PE-Cy7, PE-Vio770

780/60

735LP


21.8

54.8

633

20





3



R660_20C

APC, AF647, CF647, DiD, DRAQ5

660/20

NA


21.7

54.25



R780_60A

APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APC

780/60

735LP


21.8

54.5

Important Information

  • Baseline in use is #80998. Ensure you save new or update existing application settings after the 12th November 2018.
  • For assistance setting up an existing experiment or using application settings please book a time with flow staff.
  • Default voltages are normally good for resolving dim populations. Rule of thumb guide to settings voltages include, ensuring the rSD of the unlabelled cells is at least 2.5x the value of the rSD of electronic noise of the detector (that is viewable from the instrument configuration sheet) that accompanies each cytometer. You need to record a small file in order to accurately determine the rSD of the unlabelled cells each time. Remember this should be the minimum voltage and not the voltage to use. CST voltages are a good voltage to use with the exception that if a population is off scale, then the voltage needs to be reduced.

CantoII - ICPMR 

Baseline:  

Baseline reports: 

Cytometer Baseline Report_ICPMR.pdf

CS&T bead lot: 93308

Laser (nm)Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen2.5x rSDen

405

50





1



V450_50B

Pacific Blue, Cascade Blue, AF405, V450, BV421

450/50

NA


1640



V510_50A

CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO

510/50

502LP


16.140.25

488 

20





2



SSC

SSC

488/10

NA


N/A


B530_30E

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, SYBR Green

530/30

502LP

 

21.353.25


B585_42D

PE,PI,DsRed,DiA

585/42

556LP


22.957.25


B670_LPB

PerCP,PI, PerCP-Cy5.5,PerCP-Cy5,PE-Cy5,7AAD, BB700

670LP

655LP


22.255.5


B780_60APE-Cy7, PE-Vio770

780/60

735LP


21.152.75

633

20





3



R660_20C

APC, AF647, CF647, DiD, DRAQ5

660/20

NA


16.140.25


R780_60A

APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APC

780/60

735LP


15.639

Important Information

  • Baseline in use is #80998. Ensure you save new or update existing application settings after the 5th November 2018.
  • For assistance setting up an existing experiment or using application settings please book a time with flow staff.
  • Default voltages are normally good for resolving dim populations. Rule of thumb guide to settings voltages include, ensuring the rSD of the unlabelled cells is at least 2.5x the value of the rSD of electronic noise of the detector (that is viewable from the instrument configuration sheet) that accompanies each cytometer. You need to record a small file in order to accurately determine the rSD of the unlabelled cells each time. Remember this should be the minimum voltage and not the voltage to use. CST voltages are a good voltage to use with the exception that if a population is off scale, then the voltage needs to be reduced.

CantoII - SSM

LSRII 

Baseline:  

Baseline reports:

LSRII - Cytometer Baseline Report.pdf

LSRII - Cytometer Baseline Report.csv

CS&T bead lot: 80998

Laser (nm)Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen2.5x rSDen

488

50





1



SSC

SSC

488/10

NA


N/AN/A



B530_30B

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, SYBR Green

530/30

505LP


24.761.75


B710_50APerCP-Cy5.5, 7AAD, BB700710/50685LP
24.561.25

405

50





2



V440_40FPacific Blue, Cascade Blue, AF405, V450, BV421, Dylight405440/40NA
20.150.25


V525_50E

CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO

525/50

505LP

 

23.358.25


V610_20D

Qdot 605, BV605

610/20

600LP


35.187.75


V660_20C

Qdot 655, BV650

660/20

630LP


22.656.5


V710_50B

Qdot 700, BV711

710/50

685LP


28.471


V780_60ABV785780/60750LP
22.656.5

355

20





3



U379_28CBUV395379/28NA
17.944.75


U515_30BBUV496, DAPI, Hoechst Blue515/30450LP
16.541.25


U740_35ABUV737, Hoechst Red740/35690LP
19.248

640

40





4



R670_14CAPC, AF647, CF647, DiD, DRAQ5, SYTOX RED670/14NA
14.235.5


R730_45B

APC-R700, AF700, DRAQ5, FVS700

730/45

690LP


13.734.25


R780_60A

APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APC

780/60

750LP


14.736.75

561

100





5



Y586_15DPE, DsRed, dTomato, mTangerine, mStrawberry586/15570LP
17.744.25


Y610_20C

PE-Texas Red, mCherry, PE-CF594, AF568, PI, PE-Dazzle, 7AAD, AF594

610/20600LP
16.441


Y710_50BPE-Cy5.5, 7AAD710/50685LP
16.441


Y780_60APE-Cy7, PE-Vio770, 780/60750LP
19.147.75

Important Information

  • Baseline in use is #80998. Ensure you save new or update existing application settings after the 31st October 2018.
  • Default voltages are normally good for resolving dim populations. Rule of thumb guide to settings voltages include, ensuring the rSD of the unlabelled cells is at least 2.5x the value of the rSD of electronic noise of the detector (that is viewable from the instrument configuration sheet) that accompanies each cytometer. You need to record a small file in order to accurately determine the rSD of the unlabelled cells each time. Remember this should be the minimum voltage and not the voltage to use. CST voltages are a good voltage to use with the exception that if a population is off scale, then the voltage needs to be reduced.
  • Filter changes on the Fortessa and LSRII can be done once users have been trained. The “Non-default signage” must be displayed when using a non-default filter in place of the default.
  • For optimal detection of PE-CY5, PE-AF647 & mPlum a 670/30 filter is needed in Y710_50B with a 635LP.
  • For optimal detection of PerCP a 675/50 could be used in B710_50A with a 635LP. There are other alternatives too.
  • For optimal detection of Hoechst Blue & AF350 a 450/50 bandpass filter could be used in U515_30B, with a 425LP.
  • For optimal detection of AF633 a 670/40 bandpass filter could be used in R670_14C.

Fortessa

Baseline:  

Baseline reports:

Cytometer Baseline Report_fortessa.pdf

CS&T bead lot: 93308

Laser (nm)Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen2.5x rSDen

488

50-100

default @ 50





1



SSC

SSC

488/10

N/A


N/AN/A



B530_30B

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen

530/30

505LP


15.137.75


B710_50APerCP-Cy5.5, 7AAD, BB700710/50685LP
15.137.75

403

50





2



V440_40FPacific Blue, Cascade Blue, AF405, V450, BV421440/40N/A
10.125.25


V525_50E

CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO

525/50

505LP

 

10.125.25


V610_20D

Qdot 605, BV605

610/20

600LP


10.526.25


V660_20C

Qdot 655, BV650

660/20

630LP


11.428.5


V710_50B

Qdot 700, BV711

710/50

685LP


10.526.25


V780_60ABV785780/60750LP
1025

355

20





3



U379_28CBUV395379/28N/A
20.551.25


U515_30BBUV496, DAPI, Hoechst Blue515/30450LP
22.155.25


U740_35ABUV737, Hoechst Red740_35690LP
20.451

639

40





4



R670_14CAPC, AF647, CF647, DiD, DRAQ5670/14N/A
15.739.25


R730_45B

APC-R700,AF700, DRAQ5, FVS700

730/45

690LP


16.441


R780_60A

APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APC

780/60

750LP


1742.5

561

50





5



Y586_15DPE, DsRed, dTomato, mTangerine, mStrawberry586/15570LP
13.634


Y610_20C

PE-Texas Red, mCherry, PE-CF594, AF568, PI, PE-Dazzle

610/20600LP
14.937.25


Y710_50BPE-Cy5.5, 7AAD710/50685LP
13.634


Y780_60APE-Cy7, PE-Vio770, 780/60750LP
15.538.75

Important Information

  • Baseline in use is #80998. Ensure you save new or update existing application settings after the 31st October 2018.
  • Default voltages are normally good for resolving dim populations. Rule of thumb guide to settings voltages include, ensuring the rSD of the unlabelled cells is at least 2.5x the value of the rSD of electronic noise of the detector (that is viewable from the instrument configuration sheet) that accompanies each cytometer. You need to record a small file in order to accurately determine the rSD of the unlabelled cells each time. Remember this should be the minimum voltage and not the voltage to use. CST voltages are a good voltage to use with the exception that if a population is off scale, then the voltage needs to be reduced.
  • Filter changes on the Fortessa and LSRII can be done once users have been trained. The “Non-default signage” must be displayed when using a non-default filter in place of the default.
  • For optimal detection of PE-CY5, PE-AF647 & mPlum a 670/30 filter is needed in Y710_50B with a 635LP.
  • For optimal detection of PerCP a 675/50 could be used in B710_50A with a 635LP. There are other alternatives too.
  • For optimal detection of Hoechst Blue & AF350 a 450/50 bandpass filter could be used in U515_30B, with a 425LP.
  • For optimal detection of AF633 a 670/40 bandpass filter could be used in R670_14C.

Symphony

Baseline: 20191127 - TBU

Baseline report: TBU

CS&T bead lot: 80998

Laser ColourLaser (nm)Max power (mw)CST default power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic mirrorNotch filterLaser orderPinhole positionDetector specificsOrientationAdditional notes
Red637140140




11
5D




R820_60APC-Cy7, APC-H7, DiR, APC-FIRE820/60 BP800LP


H14121-50





R780_60spectral780/60 BP750 LP785nm

H14121-50





R730_45APC-R700, AF700, DRAQ5, FVS700730/45 BP685 LP


H11900-20





R670_30APC, AF647, CF647, DiD, DRAQ5670/30 BP665 LP


H11900-20
Schott RG665














UV355100100




23
M1&M2&M1




U820_60BUV805820/60 BP800 LP785nm

H14121-50





U780_60spectral780/60 BP770 LP


H14121-50





U740_35BUV737, Hoechst Red740/35 BP710 LP


H14121-50





U695_40
695/40685 LP









U670_30BUV661670/30 BP635 LP


H11900-20





U610_20BUV615610/20 BP600 LP


H11900-20





U586_15BUV563586/15 BP570 LP594nm

H11900-20





U515_30BUV496, DAPI, Hoechst Blue515/30 BP450 LP


H11900-20





U450_50AF350, DAPI450/50 BP410 LP


H11900-20





U379_28BUV395379/28 BP



H11900-110















Yellow561150150




34
6D




Y820_60PE-Cy7820/60 BP800 LP


H14121-50





Y780_60
780/60 BP750 LP785nm

H11900-20





Y710_50PE-Cy5.5710/50 BP685 LP


H11900-20





Y670_30PE-Cy5670/30 BP635 LP


H11900-20





Y610_20PE-CF594, PE-Texas Red, mCherry, AF568, PI, PE-Dazzle610/20 BP600 LP


H11900-20





Y586_15PE, DsRed, dTomato, mTangerine, mStrawberry586/15 BP

594nm

Y-58 Schott



H11900-40















Blue488200150




45
M1&M2




B820_60BB790820/60 BP800 LP


H14121-50





B780_60spectral780/60 BP750 LP785nm

H14121-50





B750_30BB755750/30 BP735 LP


H14121-50





B710_50BB700, PerCP-Cy5.5, 7AAD710/50 BP685 LP


H11900-20





B670_30BB660670/30 BP635 LP


H11900-20





B610_20BB630610/20 BP600 LP594nm

H11900-20





B515_30FITC, BB515, A488515/30 BP500 LP


H11900-20





SSC
488/10 BP



H11900-20















Orange594100100




56
5D




O660_20spectral660/20 BP635 LP


H14121-20





O616_23AF594, mCherry616/23 BP610 LP


H14121-20















Violet406200200




67
M1&M2&M1




V820_60BV786820/60 BP800 LP


H14121-50





V780_60spectral780/60 BP770 LP785nm 

H14121-50





V750_30BV750750/30 BP735 LP


H14121-50





V710_50BV711, Qdot700710/50 BP685 LP


H11900-20





V677_20BV650, Qdot650677/20 BP635 LP


H11900-20





V610_20BV605, Qdot605610/20 BP600 LP


H11900-20





V586_15BV570586/15 BP550 LP594nm

H11900-20





V525_50BV510, CFP, AF430, AmCyan, Cascade Yellow, V500, PO525/50 BP505 LP


H11900-20





V474_25BV480474/25 BP450 LP


H11900-20





V427_25BV421, Pacific Blue, Cascade Blue, AF405, V450,427/25 BP410 LP


H11900-20





V405_10Violet SSC405/10 BP



H11900-20







*PMT installed



H11900-20







*PMT installed



H11900-20







*PMT installed



H11900-20















IndigoNIR446_77975_10075_100




79
5D




N820_60DyLight800 (strep-avidin - recommended), AF790820/60 BP800 LP


H14121-50





I660_20spectral660/20 BP635 LP


H14121-20





I605_40spectral605/40 BP600 LP


H11900-20





I515_30spectral515/30 BP500 LP


H11900-20





I470_15CFP, Chromomycin470/15 BP450 LP


H11900-20

Symphony - SSM - December 2019

CD4 Spread Profiles - WIMR - A5.2 - CD4 Spread Profiles.pptx

BD Generated SSM - WIMR - A5.2 - SSM.xlsx

Westmead Cytometry SSM - 

Symphony Stain Index

ARIAIII - Cell Sorter

Baseline:  

Baseline reports:

70um Cytometer Baseline Report.pdf

85um Cytometer Baseline Report.pdf

100um Cytometer Baseline Report.pdf

130um Cytometer Baseline Report.pdf

CS&T bead lot: 80998

Laser (nm)Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen (70um)2.5x rSDen (70um)

633

18





1



R660_20CAPC, AF647, CF647, DiD, DRAQ5 660/20NA
8.2

20.5



R730_45BAPC-R700, AF700, DRAQ5 730/45690LP
7.118.25


R780_60AAPC-Cy7, APC-H7, DiR, APC-eFL780, APC-Fire750780/60750LP
7.118.25

488 

20





2



SSC
488/10NA
N/A




B530_30EFITC, GFP, AF488, BBYFP, 5-FAM, CFSE , BB515, ZsGreen530/30502LP
8.5

21.25



B585_42DPE, DsRed, dTomato (sub-optimal), mTangerine (sub-optimal), mStrawberry (sub-optimal)585/42556LP
8.6

21.5



B616_23CPE-Texas Red, mCherry (sub-optimal), PE-CF594, PE-AF610, AF568, PE-eFL610, PE-Dazzle594616/23610LP
9.2

23



B695_40BPerCP, PerCP-Cy5.5, 7AAD, PI, PerCP-eFL710, PerCP-Vio700, BB700695/40655LP
10

25



B780_60APE-Cy7, PE-Vio770780/60735LP
8.521.25

405

50





3



V440_40FPacific Blue, Cascade Blue, AF405, V450, DAPI, BV421, DyLight 405, eFL450, VioBlue440/40NA
7.2

18



V525_50ECFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO525/50505LP
6.6

16.5



V610_20DBV605, eFL605, Qdot 605610/20600LP
7.117.75


V660_20CBV650, eFL650, Qdot 655660/20630LP
7.719.25


V710_50BBV711, Qdot 700710/50685LP
7.218


V780_60ABV785, Qdot 800780/60750LP
6.917.25


INFLUX - Cell Sorter

Filter configuration: 

Laser (nm)

Power

(mw)

DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser order

488

50-200

default 100





1



PMT 1 - FSC (Par)Forward Scatter - Parrallel488/10NA



PMT 2 - FSC (Perp)Forward Scatter - Perpendicular488/10NA


PMT 3 - SSCSide Scatter488/10NA


PMT 4FITC, GFP, AF488, BBYFP, 5-FAM, CFSE , BB515, ZsGreen, DiO530/30600LP


PMT 5PerCP, PerCP-Cy5.5, 7AAD, PI, PerCP-eFL710, PerCP-Vio700, BB700710/50NA
405100405 nm - 100mw   2


PMT 14Pacific Blue, Cascade Blue, AF405, V450, BV421, DyLight 405, eFL450, VioBlue440/40455LP, 500LP


PMT 15CFP, eCFP480/40500LP


PMT 16CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO, VioGreen525/30550LP


PMT 17BV570, Qdot 565, 572/28585LP, 645LP


PMT 18BV605, DyLight405, Qdot 605610/20645LP


PMT 19BV650, eFL650, Qdot 655660/20685LP


PMT 20BV711, Qdot 700710/50NA
561150561 nm - 150mw   3


PMT 6PE, DsRed, dTomato, mTangerine, mStrawberry, Dil585/29600LP


PMT 7PE-Texas Red, mCherry, PE-CF594, PE-AF610, AF568, PE-eFL610, PE-Dazzle594610/20645LP


PMT 8PE-Cy5670/30685LP


PMT 9PE-Cy5.5710/50740LP


PMT 10PE-Cy7, PE-Vio770750LPNA
355100355 nm - 100mw   4


PMT 13DAPI, Hoechst Blue460/50480LP, 600LP


PMT 22BUV395379/34427LP


PMT 23BUV496520/35550LP, 600LP


PMT 24BUV661, Hoechst Red670/30NA


PMT_UnpluggedBUV563585/42NA
628120640 nm - 120mw   5


PMT 11APC, AF647, CF647, DiD, DRAQ5 670/30700LP


PMT 12AF700, DRAQ5, APC-R700740/40740LP


PMT 21APC-Cy7, APC-H7, DiR, APC-eFL780, APC-Fire750750LPNA



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