Re: PE-Cy5 on Facscalibur

Margaret Tropea (mtropea@nih.gov)
Thu, 19 Jun 1997 08:49:23 -0400

Peter,

The problem that you can have with PeCY5 results from the fact that PeCY5
is a man made tandem while PerCP is a naturally occuring tandem. Put
another way, if the energy transfer from the Pe to the CY5 is incomplete
you will get a Pe signal as well as a CY5 signal (but I think that you are
aware of this). This may be why you are having compensation problems.
Different vendors are better at making this conjugate than others. You may
want to check Volume 677 of the Annals of the New York Academy of Sciences
March 20, 1993 pp 94-112. I checked with BD instrument support and they
told me the FL3 detector was the same on both instruments.

Margaret

>Greatings all,
>
>I have been looking into the economics of our flow setup and discovered
>that I can buy PECy5 anti CD45 for much cheaper than I currently buy PerCP
>anti CD45. I proceeded to make some three color CD3/4/45 and 3/4/45 PBL's
>to do some compensation with and found that I got into real trouble :(
>
>Did something drastic go wrong with the labeling of the PBL's ??, or is
>there something I need to know about PE-Cy5 on a Calibur (ie: has anyone
>else done this easily/with difficulty ??)
>
>One problem is that I have very little quality information on the Calibur
>- the manual seems to make no mention of the of exactly which
>mirrors/filters the Calibur uses ... does it use a 650LP filter on the FL3
>detector the same as a FacsScan does ?? (if that's the case then I'm only
>just picking up the *tail* of the PE-Cy5 emission)
>
>
>Peter Chapple
>Melbourne AUSTRALIA


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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone: (765)-494-0757; FAX(765) 494-0517; Web http://www.cyto.purdue.edu , EMAIL cdrom3@flowcyt.cyto.purdue.edu