Table of Contents | ||
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CantoII - WIMR
Baseline:
Baseline reports:
CANTOII - WIMR - Cytometer Baseline Report_cantowimr.pdf
CS&T bead lot: 80998 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 |
...
CantoII - ICPMR
Baseline:
Baseline reports:
CANTOII - ICPMR - Cytometer Baseline Report.csvCANTOII - ICPMR - Cytometer Baseline Report_ICPMR.pdf
CS&T bead lot: 80998 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 |
...
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 |
...
- 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:
FORTESSA - Cytometer Baseline Report.csvFORTESSA - Cytometer Baseline Report._fortessa.pdf
CS&T bead lot: 80998 93308
Laser (nm) | Power (mw) | Detector | Common fluorochromes detectable (not exhaustive) | Band pass filter | Dichroic Mirror | Laser order | rSDen | 2.5x rSDen |
---|---|---|---|---|---|---|---|---|
488 | 50-100default @ 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 |
...
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 |
...
Laser (nm) | Power (mw) | Detector | Common fluorochromes detectable (not exhaustive) | Band pass filter | Dichroic Mirror | Laser order |
---|---|---|---|---|---|---|
488 | 50-200default 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 |
...