APC-Cy7 and PE-Cy7 compensation

Disterer, Petra (Medsch Royal Free/Medicine/Hepatology) p.disterer at medsch.ucl.ac.uk
Wed Nov 12 12:43:01 EST 2008


Dear all,

Maybe my understanding of the idea of tandem dyes is wrong, but don't APC-C7 and PE-Cy7
have the same emission spectrum seeing as Cy7 is the acceptor dye in both? So, how can
you compensate between APC-C7 and PE-Cy7?

Enlightenment would be highly appreciated.

Petra Disterer

UCL Medical School, UK



________________________________

From: Guzik, Lynda [mailto:GuziLJ at ccm.upmc.edu]
Sent: Fri 11/7/2008 16:39
To: cyto-inbox
Subject: APC-Cy7 and PE-Cy7 compensation



Hello All,

I have a researcher who would like to do 6color flow using FITC, PE, PI, APC, APC-Cy7,
and PE-Cy7.  We are using a FACSVantage DiVa with a 488nm, 635nm and UV lasers.  We are
having problems with compensation between the APC-Cy7 and PE-Cy7 channels.  Using single
color positive controls and the DiVa algorithm, the compensation looks fine.  However,
when we put APC-Cy7 and PE-Cy7 antibodies in the same tube, the plots look like there is
extreme overcompensation of PE-C7 against the APC-Cy7 channel.	I've attached a couple of
plots to illustrate.  I am able to correct for this, but wonder if I should be.  And I
fail to understand why the single color controls look fine but have a problem with the
dual stained tube.  Any input would be appreciated.

Thank you,
Lynda Guzik
McGowan Institute - Lagasse Labs
Rm 520A - Bridgeside Point Bldg
100 Technology Dr - Suite 200
Pittsburgh, PA 15219
412-235-5213
guzilj at upmc.edu<mailto:lguzilj at upmc.edu> or lyg1 at pitt.edu<mailto:lyg1 at pitt.edu>










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