[Cytometry] Staining blood before or after RBC lysis
asosink at ccf.org
Thu Aug 5 12:03:40 EDT 2010
We look at rare cells in the blood and do the staining after RBC-lysis. We tested both ways and staining after lysis works the best for our application.
I would suggest you to test both ways and then decide.
> Kewal Asosingh, M.S., Ph.D.
> Assistant Professor of Molecular Medicine
> Department of Pathobiology/NC22
> Cleveland Clinic
> 9500 Euclid Ave.
> Cleveland, OH 44195
> Phone: 216-444-0891
> Fax: 216-636-0104
> From: cytometry-bounces at lists.purdue.edu on behalf of Joanne Lannigan
> Sent: Thursday, August 5, 2010 11:44 AM
> To: Simona Zompi; cytometry at lists.purdue.edu
> Subject: Re: [Cytometry] Staining blood before or after RBC lysis
> Hi Simona:
> I think traditionally clinical labs stain in whole blood and then lyse the red cells. There are many published papers and protocols on this method. The NCCLS also has recommended procedures which you can refer to. I can't see why you would use any more antibody staining in whole blood vs. rbc lysed.
> On Thu, 5 Aug 2010 08:12:02 -0700
> Simona Zompi <simona.zompi at gmail.com> wrote:
> >Hi flowers,
> >I am setting up an assay to look at different B cell populations in the
> >blood of kids infected with dengue fever. Do you recommend to stain BEFORE
> >lysing the red blood cells or AFTER? I am currently doing it after as I use
> >less Ab (I stain in 50ul total), and I use 20ul of blood.
> >I am looking at small populations as well so I was wondering whether lysing
> >RBC before staining may induce a loss of small population or whether it's
> >the same.
> >Thank you for your help!
> >Simona ZOMPI, MD, PhD
> >Visiting Assistant Researcher
> >UC Berkeley
> >School of Public Health
> >Division of infectious diseases
> >Pr. Eva Harris lab
> >1, Barker Hall Room #416
> >Berkeley, CA 94720
> >Tel: +1 510 643 5404
> >Skype: simona_0873
> >Cytometry mailing list
> >Cytometry at lists.purdue.edu
> >Search the list archive at http://tinyurl.com/cytometry
> Joanne Lannigan, MS
> Director, Flow Cytometry Core Facility
> University of Virginia
> Jordan Hall, Room 7067
> P.O. Box 800734
> Charlottesville, VA 22908-0734
> Office: 434-924-0274
> Lab: 434-243-2695
> Fax: 434-982-1071
> email: joannelannigan at virginia.edu
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