10-color Gallios (Beckman Coulter)

 

 


 

Optical Bench

Before you decide on a fluorochrome for a flow cytometric assay, you need to know:
  • the wavelengths of the lasers in the cytometer
  • the wavelengths of the filters in front of the photomultiplier tubes (PMTs) in the cytometer
  • the absorption and fluorescence wavelengths of fluorochromes to be used
These can be simulated using the BD fluorescence spectrum viewer

450/40 PMT:

  • brilliant violet 421
  • VioBlue (Miltenyi Biotec)
  • pacific blue
  • live/dead violet fixable amine dye (Invitrogen)
  • BD Horizon V450
  • Alexa fluor 405
  • eFluor 430
  • DAPI
  • Hoechst

550/40 PMT:

  • brilliant violet 570
  • VioGreen (Miltenyi Biotec)
  • live/dead yellow fixable amine dye (Invitrogen)
  • pacific orange
  • BD Horizon V500
  • krome orange

Blue laser configuration:


525/40 PMT (B1):

  • FITC
  • alexa fluor 488
  • GFP
  • YFP
  • live/dead green fixable amine dye
  • CFSE

575/30 PMT (B2)

  • PE

620/30 PMT (B3)

  • BD Horizon PE-CF594
  • Texas Red (Invitrogen)

675/20 PMT (B4)

  • PerCP
  • PerCP-Cy5.5
  • eFluor 610
  • PI
  • 7-AAD

755LP PMT (B5)

  • PE-Cy7
  • PE-vio770

 

 

Red laser configuration:

660/20 PMT (R1)

  • APC
  • alexa fluor 647
  • eFluor 660
  • live/dead far red fixable amine dye

725/20 PMT (R2)

  • alexa fluor 700

755LP PMT (R3)

  • APC-Cy7
  • APC-H7 (BD)
  • APC-alexa fluor 750
  • APC-eFluor 780
  • APC-vio770 (Miltenyi Biotec)
  • live/dead near IR fixable amine dye