Re: Advice on 4 colour flow

From: Joseph Webster (J.Webster@centenary.usyd.edu.AU)
Date: Thu Oct 18 2001 - 17:41:21 EST


Dear Linda,
Please tell us what flow cytometer(s) you have, then we will
be better equipped to offer more advice.

On a FACScalibur, we most commonly use FITC/PE/PE-Cy5/APC
and can substitute PI or PerCP in FL3. (I believe Cychrome
is one of the proprietary names for PE-Cy5)
Those combinations are not so easy on our FACStar Plus.

You say you cannot use Cychrome and APC together, so you
must not have a FACScalibur which can compensate between FL3
and FL4...

PE-Tr may work, you would have to get a sample and test it.
PE-Cy7 is doubtful, though you just might be lucky enough to
have a super red-sensitive detector...

Regards, Joseph.

At 05:50 19/10/2001, Linda Duncan wrote:
>Dear All,
>For various reasons, I will be purchasing antibodies from
>Caltag, and need to get a combination that will work. I
>know that I can use FITC/PE/PerCP/APC but cannot use
>FITC/PE/Cychrome/APC because of the overlap in excitation
>between Cychrome and APC,but Caltag has three different
>fluorochromes on FL3: PE-TR; PE-CY5; PE-CY7. Which FL3
>Fluorochrome from Caltag will work with FITC, PE,and APC?
>More importantly which ones won't work. Please help!!
>----------------------
>Linda Duncan



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