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
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