Hi Monica, To try and help you on the topic here is a small GIF with first plot showing CD45RA-FITC and CD45RO-PE stained human lymphocytes previously gated on CD4-PECy5 positive and scatter gate. These are sorting regions around populations and on the right are plots showing sorted cells. Although there is a spread of cells (++?) between 2 populations, these gates were "selective" enough to include single positive cells only. When doing analysis one could be even more generous in setting these gates. Cheers, Sasha. ************ Dr Sasha Sreckovic Dept Path & Micro University of Bristol University Walk Bristol, BS8 1TD, UK Sasa.Sreckovic@bristol.ac.uk +44-(0)117-928-8606 ************ > From: Monica Matas <M.Matas@gmx.de> > Reply-To: "M.Matas" <M.Matas@gmx.de> > Date: Tue, 22 Aug 2000 21:35:02 +0200 > To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> > Subject: gating CD45RO+/RA+ Lymphocytes > > > > I am a student of nutrition science at the university of Bonn (Germany) and > I'm working on a study of immunophenotyping T-lymphocytes by three-color > Flow cytometry. > I want to get the percentages of the naive T-cells (CD45RA+) and memory > T-cells (CD45RO+) in the blood samples. Therefore I used the > antibodycombination of CD45RA-FITC, CD45RO-PE and CD3PerCP. > I'm experiencing difficulties by setting the gates in the dot plots. I don't > know where to set the gates for separating the two populations of cells > (naive and memory T-cells).I don´t know what criteria I have to use to > separete this two subpopulations. > (I'm using the program WinMDI 2.7) > I know there are some subpopulations between the naive and memory T-cells > like double positive T-cells (CD45RA+CD45RO+) and this makes it very > difficult to set the gates. > > I will be very obliged if you could tell me something about it or if you > could tell me about related literature. > > Thank you very much in advanced. > Monica Matas Nobis > > --------------------- > Mónica Matas Nobis > mailto:M.Matas@gmx.de > >
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