Re: Staining of Whole Blood.

PLM (plem@erols.com)
Thu, 16 Oct 1997 00:21:08 -0400

Jill,
> I never heard of staining whole blood myself until one year ago. Becton
> Dickinson and Coulter make antibodies and a lysing solution kit which
> work well.
>
> Basically, you add antibodies to 100 ul whole heparinized or EDTA blood.
> then add the 1x lysing solution (give a quick vortex before adding the
> lysing solution....works much better)at 37oF, and when transparent
> (takes 1-3 minutes in simian blood, about 30 seconds in murine blood)
> add their fixing solution. Wash 2x in DPBS, then leave in 300 ul 2&
paraformaldehyde. I'm sure this would work for human as I believe it
was designed originally for that purpose.

sorry, somehow I must have sent this prematurely.... Feel free to let me
know if have other questions!

P. Echeagaray
Flow Cytometry Services
Southern Research Frederick
echeagaray@sri.org
>
> Martin, Jill V. (Jill Martin) wrote:
> >
> > Hi to everyone in Flower Land. I have an investigator who wants
> > to look at leucocyte markers using unlysed whole blood. He says that was
> > the protocol used at the institution where he used to work. I have
> > never run samples prepared this way, nor have I heard of anyone doing it.
> > The first trial was not a success. If anyone who reads this knows if
> > and how the samples can be stained and analysed I would appreciate hearing
> > from them. I would also like to hear if you think that it is impossible
> > so I can pass that information along.
> >
> > Jill Martin, , Manager
> > Molecular Biology & Flow Cytometry Core
> > Johnson Research Building
> > Mayo Clinic Scottsdale
> >
> > 602-301-6071 (Voice)
> > 602 301-7017 (FAX)
> > email: jmartin@mayo.edu or martin.jill@mayo.edu