CantoII - WIMR

Baseline: 03 Aug 2020

Baseline reports:

Cytometer Baseline Report_cantowimr.pdf

CS&T bead lot: 93308

Laser (nm)	Power (mw)	Detector	Common fluorochromes detectable (not exhaustive)	Band pass filter	Dichroic Mirror	Laser order	rSDen	2.5x 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: 03 Aug 2020

Baseline reports:

Cytometer Baseline Report_ICPMR.pdf

CS&T bead lot: 93308

Laser (nm)	Power (mw)	Detector	Common fluorochromes detectable (not exhaustive)	Band pass filter	Dichroic Mirror	Laser order	rSDen	2.5x rSDen
405	50					1
		V450_50B	Pacific Blue, Cascade Blue, AF405, V450, BV421	450/50	NA		16	40
		V510_50A	CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO	510/50	502LP		16.1	40.25
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		21.3	53.25
		B585_42D	PE,PI,DsRed,DiA	585/42	556LP		22.9	57.25
		B670_LPB	PerCP,PI, PerCP-Cy5.5,PerCP-Cy5,PE-Cy5,7AAD, BB700	670LP	655LP		22.2	55.5
		B780_60A	PE-Cy7, PE-Vio770	780/60	735LP		21.1	52.75
633	20					3
		R660_20C	APC, AF647, CF647, DiD, DRAQ5	660/20	NA		16.1	40.25
		R780_60A	APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APC	780/60	735LP		15.6	39

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: 31 Oct 2018

Baseline reports:

LSRII - Cytometer Baseline Report.pdf

LSRII - Cytometer Baseline Report.csv

CS&T bead lot: 80998

Laser (nm)	Power (mw)	Detector	Common fluorochromes detectable (not exhaustive)	Band pass filter	Dichroic Mirror	Laser order	rSDen	2.5x rSDen
488	50					1
		SSC	SSC	488/10	NA		N/A	N/A
		B530_30B	FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, SYBR Green	530/30	505LP		24.7	61.75
		B710_50A	PerCP-Cy5.5, 7AAD, BB700	710/50	685LP		24.5	61.25
405	50					2
		V440_40F	Pacific Blue, Cascade Blue, AF405, V450, BV421, Dylight405	440/40	NA		20.1	50.25
		V525_50E	CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO	525/50	505LP		23.3	58.25
		V610_20D	Qdot 605, BV605	610/20	600LP		35.1	87.75
		V660_20C	Qdot 655, BV650	660/20	630LP		22.6	56.5
		V710_50B	Qdot 700, BV711	710/50	685LP		28.4	71
		V780_60A	BV785	780/60	750LP		22.6	56.5
355	20					3
		U379_28C	BUV395	379/28	NA		17.9	44.75
		U515_30B	BUV496, DAPI, Hoechst Blue	515/30	450LP		16.5	41.25
		U740_35A	BUV737, Hoechst Red	740/35	690LP		19.2	48
640	40					4
		R670_14C	APC, AF647, CF647, DiD, DRAQ5, SYTOX RED	670/14	NA		14.2	35.5
		R730_45B	APC-R700, AF700, DRAQ5, FVS700	730/45	690LP		13.7	34.25
		R780_60A	APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APC	780/60	750LP		14.7	36.75
561	100					5
		Y586_15D	PE, DsRed, dTomato, mTangerine, mStrawberry	586/15	570LP		17.7	44.25
		Y610_20C	PE-Texas Red, mCherry, PE-CF594, AF568, PI, PE-Dazzle, 7AAD, AF594	610/20	600LP		16.4	41
		Y710_50B	PE-Cy5.5, 7AAD	710/50	685LP		16.4	41
		Y780_60A	PE-Cy7, PE-Vio770,	780/60	750LP		19.1	47.75

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: 05 Aug 2020

Baseline reports:

Cytometer Baseline Report_fortessa.pdf

CS&T bead lot: 93308

Laser (nm)	Power (mw)	Detector	Common fluorochromes detectable (not exhaustive)	Band pass filter	Dichroic Mirror	Laser order	rSDen	2.5x rSDen
488	50-100 default @ 50					1
		SSC	SSC	488/10	N/A		N/A	N/A
		B530_30B	FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen	530/30	505LP		15.1	37.75
		B710_50A	PerCP-Cy5.5, 7AAD, BB700	710/50	685LP		15.1	37.75
403	50					2
		V440_40F	Pacific Blue, Cascade Blue, AF405, V450, BV421	440/40	N/A		10.1	25.25
		V525_50E	CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO	525/50	505LP		10.1	25.25
		V610_20D	Qdot 605, BV605	610/20	600LP		10.5	26.25
		V660_20C	Qdot 655, BV650	660/20	630LP		11.4	28.5
		V710_50B	Qdot 700, BV711	710/50	685LP		10.5	26.25
		V780_60A	BV785	780/60	750LP		10	25
355	20					3
		U379_28C	BUV395	379/28	N/A		20.5	51.25
		U515_30B	BUV496, DAPI, Hoechst Blue	515/30	450LP		22.1	55.25
		U740_35A	BUV737, Hoechst Red	740_35	690LP		20.4	51
639	40					4
		R670_14C	APC, AF647, CF647, DiD, DRAQ5	670/14	N/A		15.7	39.25
		R730_45B	APC-R700,AF700, DRAQ5, FVS700	730/45	690LP		16.4	41
		R780_60A	APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APC	780/60	750LP		17	42.5
561	50					5
		Y586_15D	PE, DsRed, dTomato, mTangerine, mStrawberry	586/15	570LP		13.6	34
		Y610_20C	PE-Texas Red, mCherry, PE-CF594, AF568, PI, PE-Dazzle	610/20	600LP		14.9	37.25
		Y710_50B	PE-Cy5.5, 7AAD	710/50	685LP		13.6	34
		Y780_60A	PE-Cy7, PE-Vio770,	780/60	750LP		15.5	38.75

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 Colour	Laser (nm)	Max power (mw)	CST default power (mw)	Detector	Common fluorochromes detectable (not exhaustive)	Band pass filter	Dichroic mirror	Notch filter	Laser order	Pinhole position	Detector specifics	Orientation	Additional notes
Red	637	140	140						1	1		5D
				R820_60	APC-Cy7, APC-H7, DiR, APC-FIRE	820/60 BP	800LP				H14121-50
				R780_60	spectral	780/60 BP	750 LP	785nm			H14121-50
				R730_45	APC-R700, AF700, DRAQ5, FVS700	730/45 BP	685 LP				H11900-20
				R670_30	APC, AF647, CF647, DiD, DRAQ5	670/30 BP	665 LP				H11900-20		Schott RG665

UV	355	100	100						2	3		M1&M2&M1
				U820_60	BUV805	820/60 BP	800 LP	785nm			H14121-50
				U780_60	spectral	780/60 BP	770 LP				H14121-50
				U740_35	BUV737, Hoechst Red	740/35 BP	710 LP				H14121-50
				U695_40		695/40	685 LP
				U670_30	BUV661	670/30 BP	635 LP				H11900-20
				U610_20	BUV615	610/20 BP	600 LP				H11900-20
				U586_15	BUV563	586/15 BP	570 LP	594nm			H11900-20
				U515_30	BUV496, DAPI, Hoechst Blue	515/30 BP	450 LP				H11900-20
				U450_50	AF350, DAPI	450/50 BP	410 LP				H11900-20
				U379_28	BUV395	379/28 BP					H11900-110

Yellow	561	150	150						3	4		6D
				Y820_60	PE-Cy7	820/60 BP	800 LP				H14121-50
				Y780_60		780/60 BP	750 LP	785nm			H11900-20
				Y710_50	PE-Cy5.5	710/50 BP	685 LP				H11900-20
				Y670_30	PE-Cy5	670/30 BP	635 LP				H11900-20
				Y610_20	PE-CF594, PE-Texas Red, mCherry, AF568, PI, PE-Dazzle	610/20 BP	600 LP				H11900-20
				Y586_15	PE, DsRed, dTomato, mTangerine, mStrawberry	586/15 BP		594nm Y-58 Schott			H11900-40

Blue	488	200	150						4	5		M1&M2
				B820_60	BB790	820/60 BP	800 LP				H14121-50
				B780_60	spectral	780/60 BP	750 LP	785nm			H14121-50
				B750_30	BB755	750/30 BP	735 LP				H14121-50
				B710_50	BB700, PerCP-Cy5.5, 7AAD	710/50 BP	685 LP				H11900-20
				B670_30	BB660	670/30 BP	635 LP				H11900-20
				B610_20	BB630	610/20 BP	600 LP	594nm			H11900-20
				B515_30	FITC, BB515, A488	515/30 BP	500 LP				H11900-20
				SSC		488/10 BP					H11900-20

Orange	594	100	100						5	6		5D
				O660_20	spectral	660/20 BP	635 LP				H14121-20
				O616_23	AF594, mCherry	616/23 BP	610 LP				H14121-20

Violet	406	200	200						6	7		M1&M2&M1
				V820_60	BV786	820/60 BP	800 LP				H14121-50
				V780_60	spectral	780/60 BP	770 LP	785nm			H14121-50
				V750_30	BV750	750/30 BP	735 LP				H14121-50
				V710_50	BV711, Qdot700	710/50 BP	685 LP				H11900-20
				V677_20	BV650, Qdot650	677/20 BP	635 LP				H11900-20
				V610_20	BV605, Qdot605	610/20 BP	600 LP				H11900-20
				V586_15	BV570	586/15 BP	550 LP	594nm			H11900-20
				V525_50	BV510, CFP, AF430, AmCyan, Cascade Yellow, V500, PO	525/50 BP	505 LP				H11900-20
				V474_25	BV480	474/25 BP	450 LP				H11900-20
				V427_25	BV421, Pacific Blue, Cascade Blue, AF405, V450,	427/25 BP	410 LP				H11900-20
				V405_10	Violet SSC	405/10 BP					H11900-20
						*PMT installed					H11900-20
						*PMT installed					H11900-20
						*PMT installed					H11900-20

IndigoNIR	446_779	75_100	75_100						7	9		5D
				N820_60	DyLight800 (strep-avidin - recommended), AF790	820/60 BP	800 LP				H14121-50
				I660_20	spectral	660/20 BP	635 LP				H14121-20
				I605_40	spectral	605/40 BP	600 LP				H11900-20
				I515_30	spectral	515/30 BP	500 LP				H11900-20
				I470_15	CFP, Chromomycin	470/15 BP	450 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: 09 Nov 2018

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)	Detector	Common fluorochromes detectable (not exhaustive)	Band pass filter	Dichroic Mirror	Laser order	rSDen (70um)	2.5x rSDen (70um)
633	18					1
		R660_20C	APC, AF647, CF647, DiD, DRAQ5	660/20	NA		8.2	20.5
		R730_45B	APC-R700, AF700, DRAQ5	730/45	690LP		7.1	18.25
		R780_60A	APC-Cy7, APC-H7, DiR, APC-eFL780, APC-Fire750	780/60	750LP		7.1	18.25
488	20					2
		SSC		488/10	NA		N/A
		B530_30E	FITC, GFP, AF488, BBYFP, 5-FAM, CFSE , BB515, ZsGreen	530/30	502LP		8.5	21.25
		B585_42D	PE, DsRed, dTomato (sub-optimal), mTangerine (sub-optimal), mStrawberry (sub-optimal)	585/42	556LP		8.6	21.5
		B616_23C	PE-Texas Red, mCherry (sub-optimal), PE-CF594, PE-AF610, AF568, PE-eFL610, PE-Dazzle594	616/23	610LP		9.2	23
		B695_40B	PerCP, PerCP-Cy5.5, 7AAD, PI, PerCP-eFL710, PerCP-Vio700, BB700	695/40	655LP		10	25
		B780_60A	PE-Cy7, PE-Vio770	780/60	735LP		8.5	21.25
405	50					3
		V440_40F	Pacific Blue, Cascade Blue, AF405, V450, DAPI, BV421, DyLight 405, eFL450, VioBlue	440/40	NA		7.2	18
		V525_50E	CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO	525/50	505LP		6.6	16.5
		V610_20D	BV605, eFL605, Qdot 605	610/20	600LP		7.1	17.75
		V660_20C	BV650, eFL650, Qdot 655	660/20	630LP		7.7	19.25
		V710_50B	BV711, Qdot 700	710/50	685LP		7.2	18
		V780_60A	BV785, Qdot 800	780/60	750LP		6.9	17.25

INFLUX - Cell Sorter

Filter configuration:

Laser (nm)	Power (mw)	Detector	Common fluorochromes detectable (not exhaustive)	Band pass filter	Dichroic Mirror	Laser order
488	50-200 default 100					1
		PMT 1 - FSC (Par)	Forward Scatter - Parrallel	488/10	NA
		PMT 2 - FSC (Perp)	Forward Scatter - Perpendicular	488/10	NA
		PMT 3 - SSC	Side Scatter	488/10	NA
		PMT 4	FITC, GFP, AF488, BBYFP, 5-FAM, CFSE , BB515, ZsGreen, DiO	530/30	600LP
		PMT 5	PerCP, PerCP-Cy5.5, 7AAD, PI, PerCP-eFL710, PerCP-Vio700, BB700	710/50	NA
405	100	405 nm - 100mw				2
		PMT 14	Pacific Blue, Cascade Blue, AF405, V450, BV421, DyLight 405, eFL450, VioBlue	440/40	455LP, 500LP
		PMT 15	CFP, eCFP	480/40	500LP
		PMT 16	CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO, VioGreen	525/30	550LP
		PMT 17	BV570, Qdot 565,	572/28	585LP, 645LP
		PMT 18	BV605, DyLight405, Qdot 605	610/20	645LP
		PMT 19	BV650, eFL650, Qdot 655	660/20	685LP
		PMT 20	BV711, Qdot 700	710/50	NA
561	150	561 nm - 150mw				3
		PMT 6	PE, DsRed, dTomato, mTangerine, mStrawberry, Dil	585/29	600LP
		PMT 7	PE-Texas Red, mCherry, PE-CF594, PE-AF610, AF568, PE-eFL610, PE-Dazzle594	610/20	645LP
		PMT 8	PE-Cy5	670/30	685LP
		PMT 9	PE-Cy5.5	710/50	740LP
		PMT 10	PE-Cy7, PE-Vio770	750LP	NA
355	100	355 nm - 100mw				4
		PMT 13	DAPI, Hoechst Blue	460/50	480LP, 600LP
		PMT 22	BUV395	379/34	427LP
		PMT 23	BUV496	520/35	550LP, 600LP
		PMT 24	BUV661, Hoechst Red	670/30	NA
		PMT_Unplugged	BUV563	585/42	NA
628	120	640 nm - 120mw				5
		PMT 11	APC, AF647, CF647, DiD, DRAQ5	670/30	700LP
		PMT 12	AF700, DRAQ5, APC-R700	740/40	740LP
		PMT 21	APC-Cy7, APC-H7, DiR, APC-eFL780, APC-Fire750	750LP	NA

Cytometer Configurations

CantoII - WIMR

405

50

488

20

633

20

CantoII - ICPMR

405

50

488

20

633

20

CantoII - SSM

LSRII

488

50

405

50

355

20

640

40

561

100

Fortessa

488

50-100

403

50

355

20

639

40

561

50

Symphony

Symphony - SSM - December 2019

Symphony Stain Index

ARIAIII - Cell Sorter

633

18

488

20

405

50

INFLUX - Cell Sorter

488

50-200

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