qdots and fixative problems

Perry, Greg gap00438 at creighton.edu
Tue Nov 18 15:14:54 EST 2008


Katie,

We also use Qdots in 7-10 color phenotyping panels and fix in 1% PFA.
We do NOT wash off the fixative before running the samples, and we have
never seen a loss of Qdot signal after storage (up to 5 days).	We have
a FACSAria and run Qdots on our UV laser.  We do notice an increase in
autofluorescence in the UV channels after several days of storage (more
so than on signals from the blue or red lasers), but not a decrease in
specific antigen fluorescence. 


- Greg
Greg A. Perry, Ph.D.
Technical Director
Flow Cytometry Core Facility
Creighton University
Omaha, NE  68178
(402) 280-1841

-----Original Message-----
From: Williams, Katie Lynn [mailto:KWILLIAMS20 at PARTNERS.ORG] 
Sent: Tuesday, November 18, 2008 7:43 AM
To: cyto-inbox
Subject: qdots and fixative problems

Hi,
I wanted to know if anyone has had success fixing their samples stained
with
qdots.	We have been using commercial qdots and have had problems
finding an
appropriate fixation method that does not drastically decrease the
signal if washed off.  We usually use 1% PFA, however after fixation, if
the sample is washed we lose our qdot populations.  So far, we are just
using them in our phenotyping panels, but this definitely poses a
problem if we want to utilize them in our ICS panels.  Any suggestions
would be greatly appreciated!

Thanks,
Katie

Katie L. Williams, Lab Technician
University of KwaZulu Natal
Nelson R Mandela School of Medicine
HIV Pathogenesis Programme
DDMRI 1st Floor Room 114
719 Umbilo Road, Congella 4013
KwaZulu Natal South Africa





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