PE-Cy7 and compensation problems
Fischer, Randy (NIH/NIAMS) [E]
fischer1 at mail.nih.gov
Sat Apr 14 21:41:48 EDT 2007
This sounds like multiple problems. Starting with the second problem, the PE and PE-Cy7,
this should not be an issue unless your tandem is bad. We, and others, routinely use PE
and PE-Cy7 without any issues. This really sounds like your PE-Cy7 has become
dissociated with the PE now being the only fluorochrome left.
With the problem of APC in your PE-Cy7, how much spillover? I would expect very little
to none normally for two reasons-different light paths via the fiber optics from the two
different lasers and very different emission curves. Off the top of my head this sounds
like something misaligned-have BD check it out.
By the way, I have used the BD 34 PE-Cy7 and not had problems.
From: Virginia Portillo [mailto:Virginia.Portillo at manchester.ac.uk]
Sent: Thu 4/12/2007 11:48 AM
Subject: PE-Cy7 and compensation problems
I would like to do some multicolor experiments in a FACS Aria using the folowing
CD34 PE-Cy7 tandem dye
monoclonal Ab detected by a APC- conj. sec Ab
We have tryed a similar experimet in the past and had some problems using the
CD34 PE-Cy7 Ab such as:
-APC leaking on the PE-Cy7 channel
-when using PE together with PE-Cy7, could not compensate either.
All comercial Abs I use are BD so they do not differ on fluorochrome:protein
I am thinking on changing my CD34 or secondary Abs but for example the emission
of CD34 Cy5.5-PerCP would overlap with APC.
I would appreciate your comments on using tandem dyes in particular PE-Cy7
conjugates and/or any suggestions about using different color combinations on
my experiment to avoid these problems.
Dr Virginia Portillo
The University of Manchester
Manchester Interdisciplinary Biocentre
131 Princess Street
Tel: 0161 306 5820
Fax: 0161 236 0409
More information about the Cytometry