Re: 4 colours on FACS Calibur

From: Margaret Tropea (mtropea@nih.gov)
Date: Mon Nov 29 1999 - 08:51:37 EST


An even bigger issue is that the PE-CY5 is chemically synthesized and
therefore is not uniform in its energy transfer (or PE leakage as referred
to by Carleton Stewart) from manufacturer to manufacturer.  This can make
compensation a real nightmare (see Communications in Clinical Cytometry vol
38, no 4, p161)  PerCP is a naturally occurring compound and does not
suffer from this inconsistency.
Margaret

On 11/28 Michael Ormerod wrote:

I have not worked on this system myself but I am told that the He-Ne laser
>will excite the Cy5 on the PE-Cy5 conjugate directly, which accounts for
>the need to apply so much compensation. Looking at the excitation/emission
>spectra, this is what would be expected.
>
>I think that it might be best to use PerCP as the third dye.
>
>Michael Ormerod
>34 Wray Park Road
>Reigate RH2 ODE
>Telephone: +44 (0)1737 241726
>PLEASE NOTE NEW FAX & MOBILE NUMBERS
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>Web site: http://ourworld.compuserve.com/homepages/Michael_Ormerod

>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>
Margaret M. Tropea
Critical Care Medicine Department
National Institutes of Health
(ph) 301-496-7752
(fax) 301-480-3389
>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>



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