8-color MACSQuant VYB (Miltenyi Biotec)

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

Violet laser configuration:

450/50 PMT:

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

525/50 PMT:

  • brilliant violet 510
  • VioGreen (Miltenyi Biotec)
  • vGFP
  • live/dead aqua fixable amine dye (Invitrogen)
  • pacific orange
  • BD Horizon V500
  • AmCyan

Blue laser configuration:


525/50 PMT (B1)

  • FITC
  • alexa fluor 488
  • GFP
  • YFP

614/50 PMT (B2)

  • PI
  • 7-AAD
  • Lss-mKate
  • YFP

Yellow laser configuration:


586/15 PMT (Y1)

  • PE
  • yellow live/dead fixable amine dye

615/20 PMT (Y2)

  • mCherry
  • dsRed
  • tdTomato
  • Texas Red
  • BD Horizon PE-CF594

661/20 PMT (Y3)

  • PE-Cy5
  • PE-Cy5.5
  • APC
  • mKate
  • far red live/dead fixable amine dye

750LP PMT (Y4)

  • PE-Cy7
  • PE-vio770 (Miltenyi Biotec)
  • PE-alexa fluor 750
  • APC-Cy7
  • APC-H7
  • APC-eFluor 780
  • APC-alexa fluor 750
  • APC-vio770 (Miltenyi Biotec)