Avis- We routinely use a 5 color set-up on our Elite of FITC/PE/ECD/PerCP/APC using the 488 and HeNe lasers, so it is possible to use FITC/PE/PerCP/APC on the Elite as well as the Calibur. Be aware that you can not use any of the PE-Cy5 conjugates (Cychrome, etc) with the HeNe in conjunction with APC as you will get results that are falsely double positive (i.e. anything that is positive for PE-Cy5 will also appear positive for APC, whether it really is or not) because the HeNe will directly excite the Cy5 portion of the conjugate. I suppose it may be possible to color compensate this out but I suspect this is more of a nightmare than you want to deal with. cheers- Regina >>> Tom Mc Closkey <thomasm@nshs.edu> 11/07/00 12:21PM >>> Avis Peters wrote: > > We are looking into performing 4-color immunophenotyping analysis > (leukemia/lymphoma) that must be compatable on a Coulter Elite and BD Calibur. > We want to use FITC,PE, and APC and are wondering which would be the best 3rd > color-PE/Cy7, Percp/Cy5.5, APC/Cy7, ECD, or other fluorochomes? It will be difficult to find get a 4 color combo that will work on both machines. Thae Calibur is designed for FITC/PE/perCP/APC Regina H. Harley, M.S. Lab Supervisor - Flow Cytometry Facility Center for Vaccine Development University of Maryland - Baltimore 685 West Baltimore Street Baltimore, MD 21201 (410) 706-7376 FAX: (410) 706-6205 rharley@medicine.umaryland.edu
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