[Cytometry] whole blood

Nandita Hazra drnhazra at gmail.com
Wed Feb 19 05:29:57 EST 2014


Dear Barbara,

We had some trouble with the BD FACS lyse solution too, especially if it is
diluted to the working solution and not used up within 48h. But we found a
working solution to this issue- Vortexing the blood well once after adding
FACS Lyse does make a difference and sometimes, even keeping the tube with
the FACS lyse solution about 10 min longer than the listed time in the
protocol.
When we encountered this problem, we spoke to the BD representative as well
as standardized the volume of lysing solution, time of lysis as well as
tried other steps such as vortexing while using whole blood from a healthy
control, not wanting to waste effort on rare SSPE samples that I am
currently working on.

Hope this was useful,
Nandita


On Tue, Feb 18, 2014 at 9:47 PM, Marta Mesa <mmesa at javeriana.edu.co> wrote:

> Hi Barbara,
>
> OptiLyse® BLysing Solution Beckman-Coulter PN IM1400 works quite well.
> I have used it with nice results in the  phenotyping of circulating
> leukocytes from children and adults with no lyse no wash protocol.
>
> Best regards,
>
> Marta Mesa
> Bogotá, Colombia
>
>
>
> On Feb 7, 2014, at 4:03 AM, barbara morandi wrote:
>
> > Dear all,
> > I would like to know whether someone can suggest how to perform
> immunofluorescence staining on whole blood (CD3, CD56, CD16, CD19, CD14).
> I've tried to lyse red cells with BD FACS lysis solution but it seems it
> doesn't work very well. Can anyone suggest a protocol to efficiently lyse
> red cells?
> > Thanks a lot,
> > Barbara
> >
> > Dr. Barbara Morandi
> > Immunology Unit
> > Istituto Nazionale per la Ricerca sul Cancro
> > CBA- L.go R.Benzi, 10
> > 16132 - Genova, Italy
> > Tel :39 (010) 5558-221
> > Fax: 39 (010) 354282
> > E-mail:
> morandibarb at yahoo.it_______________________________________________
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>
>
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>


-- 
Lt Col (Dr) Nandita Hazra
PSG FAIMER 2010 Fellow
PhD Scholar
Dept of Neurovirology
NIMHANS
Bengaluru 560029


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