We have been using such approach without a problem, in a machine similar to the one you have. CD4 APC stained cells are nicely separated concurrently with PI and Annexin V staining in FL2 and FL1, respectively. Esper Kallas -----Mensagem original----- De: Nealley, Eric W Mr USAMRICD [mailto:Eric.Nealley@apg.amedd.army.mil] Enviada em: quarta-feira, 21 de novembro de 2001 12:55 Para: Cytometry Mailing List Assunto: 3 color combination? 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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