Interesting thought Eric. I've been using our 4 color FACSCalibur for about 6 months with APC CD61, FITC annexin V and RPE-CD42 in roughly the same setup and not had any problems. Neither of the latter 2 though emit in the FL4 bandpass though. We use PI nuclear staining for WBC count and were thinking of using the APC channel for some further labels so I'm interested to hear of any conflict. Craig ---------- From: Eric.Nealley@apg.amedd.army.mi To: cyto-inbox Subject: 3 color combination? Date: Thursday, November 22, 2001 1:27AM We are trying to run a 3-color experiment on our FACScalibur (with FL4 option) and I was concerned about spectral overlap between propidium iodide and APC. We want to use annexin V FITC, propidium iodide and CD3 APC. I know there is a time delay procedure to calibrate the instrument, but does the spectral overlap between propidium iodide and APC get "cancelled out" by the time delay? Our instrument setup: FL1: 530/30, FL2: 585/42, FL3: 670/LP and FL4: 661/16. Thanks in advance. Eric Nealley
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