gating CD45RO+/RA+ Lymphocytes

Dr Sasha Sreckovic sasa.sreckovic at bris.ac.uk
Tue Aug 29 05:37:58 EST 2000


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 at bristol.ac.uk
+44-(0)117-928-8606
************



> From: Monica Matas <M.Matas at gmx.de>
> Reply-To: "M.Matas" <M.Matas at gmx.de>
> Date: Tue, 22 Aug 2000 21:35:02 +0200
> To: Cytometry Mailing List <cytometry at 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 at gmx.de
> 
> 

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