Common Flow Cytometry Fluorochrome Combinations

Standard or reasonably common fluorochrome combinations used in WEHI instruments:

FACStar+ Laser and Fluorochrome Options:

Laser 1 488nm FL1 FITC , GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
FL2** PE, Cy3, RFP
FL3* PI, 7AAD, PE.Tx , PE.Cy5, PerCp.Cy5.5, PE.Cy7
Laser 2 561nm FL3* APC, Cy5, Alexa630, Alexa647, Alexa660,
FL4** PE, dTomato, Tx.Red, Alexa594, mCherry
  • * FL3 may accept laser1 *OR* laser2-excited signals.
  • PE.Cy5 should not be combined with any FL3/laser 2 fluorochrome.
  • PE.Tx should not be combined with any FL4 fluorochrome.
  • **TEMPORARY Nov2011: PE.488 is incompatible with Tex/Alexa594/mCherry unless used as dump.

 

DiVa Laser and Fluorochrome Options:

Laser 1 488nm FL1 FITC , GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
FL2 PE, Cy3, RFP
FL3 7AAD, PE.Cy5, PerCp.Cy5.5, PE.Cy7
FL6 PI, PE.Tx, PE.Alexa610
515nm* FL2 TRITC
Laser 2 605nm FL4 Tx.Red, Alexa594, APC.Cy7*, Alexa680*
FL5 APC, Cy5, Alexa630, Alexa647, Alexa660
550-650nm FL4/FL5 dye laser tuned for special fluorochrome excitation
Laser 3 351-364nm FL7 Hoechst, Alexa350, Cascade Blue, AMCA, Indo-1, DAPI, Qdot605**
FL8 Indo-1, CascadeYellow, FluoroGold, HoechstRed (SP cells), Qdot655*
  • * Allow 15 minutes setup time
  • ** reduced sensitivity due to dye laser glow
  • PE.Cy5 should not be combined with any FL5 fluorochrome
  • PE.Tx should not be combined with any FL4 fluorochrome

 

MoFlo Upgrade Laser and Fluorochrome Options:

Laser 1 488nm FL1 FITC , GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
FL2 PE, Cy3, RFP
FL3 PI, PE.Tx, PE.Alexa610
FL4 PE.Cy5, PerCp.Cy5.5, PE.Cy7, 7AAD
Laser 2 641nm AND/OR 561nm
 FL5 APC.Cy7 OR APC OR Alexa594, mCherry
FL6 APC, Cy5, Alexa630, Alexa647, Alexa660 OR Alexa594 OR PE, dTomato
Laser 3 351-364nm AND/OR 405nm
 FL7 Hoechst, Alexa350, Cascade Blue, AMCA, Indo-1, DAPI, Qdot605*, Alexa405*, Pacific Blue*
FL8 Indo-1, CascadeYellow, FluoroGold, HoechstRed (SP cells), Qdot655*
  • * Allow 15 minutes setup time
  • PE.Cy5 should not be combined with any FL5 fluorochrome

 

Aria-C Laser and Fluorochrome Options:

Laser 1 488nm E FITC, GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
D PE, Cy3, RFP
C PI, PE.Tx, PE.Alexa610, 7AAD, PE.Cy5
B PerCp.Cy5.5
A PE.Cy7
Laser 2 405nm E Alexa 405, Pacific Blue
D 525nm
C CascadeYellow
B 585nm
A QDot655
Laser 3 375nm C Hoechst Blue, Alexa350, Cascade Blue, AMCA, DAPI
B FluoroGold
A Hoechst Red (SP cells)
Laser 4 635nm B APC, Cy5, Alexa630, Alexa647
A APC.Cy7, Alexa 680, Alexa 700

 

Aria-W Upgrade Laser and Fluorochrome Options (Not yet in full service.  Enquire for details):

Laser 1 488nm E FITC, GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
D PE, Cy3, RFP
C PI, PE.Tx, PE.Alexa610, 7AAD, PE.Cy5
B PerCp.Cy5.5
A PE.Cy7
Laser 2 405nm AND 375nm
 E Alexa 405, Pacific Blue, Hoechst Blue, Alexa350, Cascade Blue, AMCA, DAPI
D 525nm, FluoroGold
C CascadeYellow
B 585nm
A QDot655 OR Hoechst Red (SP cells)
Laser 3 561nm C PE, dTomato
B Alexa594, mCherry
A PE.Cy7
Laser 4 641nm B APC, Cy5, Alexa630, Alexa647
A APC.Cy7, Alexa 680, Alexa 700

 

inFlux Laser and Fluorochrome Options:

Laser 1 488nm PMT 1 FSC
PMT 2 SSC
PMT 3 530/40 FITC
PMT 4 670/30 PI
PMT 5 710/50 7AAD, PerCp.Cy5.5
Laser2 405nm PMT 6 460/50 PacificBlue
PMT 7 520/35 AmCyan
PMT 8 610/20 QDot605
PMT 9 660/20 QDot655
PMT 10 710/50 QDot705
PMT 17 750LP QDot750
Laser 3 UV-355nm PMT 16 460/50 Hoechst blue SP
PMT 20 670/30 Hoechst red SP
Laser 4 561nm PMT 11 585/29 PE
PMT 12 610/20 PE.Tx, Alexa590, mCherry
PMT 13 670/30 PE.Cy5
PMT 18 750LP PE.Cy7
Laser 5 640nm PMT 14 670/30 APC
PMT 15 720/40 Alexa680
PMT 19 750LP APC.Cy7

 

LSR1 Laser and Fluorochrome Options:

Laser 1 488nm FL1 FITC , GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
FL2 PE, Cy3, RFP
FL3 PI, 7AAD, PE.Tx , PE.Cy5, PerCp.Cy5.5, PE.Cy7
Laser 2 325nm FL4 Indo-1, CascadeYellow, FluoroGold
FL5 Hoechst, Alexa350, Cascade Blue, AMCA, Indo-1, DAPI
Laser 3 632nm FL6/SSC-W APC, Cy5, Alexa630, Alexa647
FL3* Alexa680, APC.Cy7
  • * Special setup for laser 3 FL3 excitation; not commonly used
  • PE.Cy5 should not be combined with any FL6 fluorochrome

 

LSR II-C Laser and Fluorochrome Options:

Laser 1 488nm E FITC, GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
D PE, Cy3, RFP
C PI, PE.Tx, PE.Alexa610
B 7AAD, PerCp.Cy5.5, PE.Cy5
A PE.Cy7
Laser 2 355nm D Hoechst Blue, INDO-1 blue, Alexa 350
C Indo-1 green, Cascade Yellow, FluoroGold
B QDot605
A QDot655, Hoechst Red (SP cells)
Laser 3 633nm B APC, Cy5, Alexa630, Alexa647
A APC.Cy7, Alexa 680, Alexa 700

 

LSR II-W Upgrade Laser and Fluorochrome Options:

Laser 1 488nm E FITC, GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
D PE, Cy3, RFP
C PI, PE.Tx, PE.Alexa610
B 7AAD, PerCp.Cy5.5, PE.Cy5
A PE.Cy7
Laser2 561nm B PE, dTomato
A Alexa594, mCherry
Not installed
 PE.Cy7
Laser 3 355nm D Hoechst Blue, INDO-1 blue, Alexa 350
C Indo-1 green, Cascade Yellow, FluoroGold
B QDot605
A QDot655, Hoechst Red (SP cells)
Laser 4 633nm B APC, Cy5, Alexa630, Alexa647
A APC.Cy7, Alexa 680, Alexa 700

 

Fortessa Laser and Fluorochrome Options:

Laser 1 488nm E SSC
D FITC, GFP, CFSE, 5'6Carboxyfluorescein, Alexa488
C PE**, Cy3
B PI, PE.Tx
A 7AAD, PerCp.Cy5.5
Laser2 405nm F PacificBlue
E AmCyan, Fluorogold*
D QDot565
C QDot585
B QDot605
A QDot655
Laser 3 640nm C APC
B Alexa 680, Alexa 700
A APC.Cy7
Laser 4 561nm E
 PE**, dTomato
D
 PE.Tx, Alexa590, mCherry
C PE.Cy5
B PE.Cy5.5
A PE.Cy7
  • * Fluorogold less well excited at 405nm than with UV.
  • **PE is better excited at 561nm which also results in no spillover from FITC, CFSE..

 

FACSCalibur Laser and Fluorochrome Options:

Laser 1 488nm FL1 FITC , GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
FL2 PE, Cy3, RFP
FL3 PI, 7AAD, PE.Tx , PE.Cy5, PerCp.Cy5.5, PE.Cy7
Laser 2 635nm FL4 APC, Cy5, Alexa630, Alexa647, Alexa660
  • PE.Cy5 should not be combined with any FL4 fluorochrome

 

FACSCan 1 and 2 Laser and Fluorochrome Options:

Laser 1 488nm FL1 FITC , GFP, CFSE, 5'6Carboxyfluorescein, Alexa488, YFP
FL2 PE, Cy3, RFP
FL3 PI, 7AAD, PE.Tx , PE.Cy5, PerCp.Cy5.5, PE.Cy7

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