Re: three color analysis

Florence Harrod (fharrod@pharmingen.com)
11 Dec 1995 10:39:48 U

Reply to: RE>>three color analysis
Why not use PE for the second color? We use it routinely, every day, on
FACscans with FITC and PI. The only time there is a problem between the PE and
PI is when the PE is so bright, greater than three logs, that it cannot be
compensated out from the PI channel.

--------------------------------------
Date: 12/8/95 6:01 PM
To: cyto-inbox
From: Zawadzki, Jerome A.

No question about it Ned. That is going to be a very difficult combination
to work with. The efficiency of Cy3 at 488 is one problem and the PI spill
over into fl2 is the other. If your Cy3 signal is strong you may be able
compensate the PI. I would guess that the PI is for dead /live
discrimination. Have you tried the Cy3 on the instrument yet? To answer your
question about getting around this. Drop the Cy3 if you have that option. If
you have to use PI then you might try Fitc and Percp. Just some thoughts
Ned. Good Luck . Jerome
----------
From: owner-cyto-sendout
To: cyto-inbox
Subject: three color analysis
Date: Wednesday, December 06, 1995 12:05PM

I would like to do three color analysis on our FACSCAN using FITC, Cy3 and
propidium iodide. I'm told that there is too much spill over of the PI
into FL2, such that I would not get reliable results on the Cy3 staining.
Is this true. Is there anyway around this problem? Thanks. Ned Landau,
Aaron Diamond AIDS Research Center.

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From: "Zawadzki, Jerome A." <jzawadzki@molecular.princeton.edu>
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Subject: RE: three color analysis
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