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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