Mikayla, patients post ablative marrow radiation therapy will have very little normally staining cells. On one such patient this evening I was able to get 27 lymphocytes out of 10^5 events. So I wouldn't complain about 50% :) If your patients aren't in that group of recovering radiation patients, I would check the antibody titrations etc.. I'm assuming your normal controls are staining OK? -maciej --- Mikayla Kob <kob@cbr.med.harvard.edu> wrote: > Dear cytometry community, > > We have been establishing a panel to phenotype lymphocytes from > clinical peripheral blood samples. We have been using a whole blood > staining and lysis procedure. The samples that I have received so > far have had an unusually large percentage of CD45- cells (over > 50%) in the lymphocyte gate as determined by forward and side > scatter. These cells are also CD3, 4, 8, and 19 negative, appear > to be CD43 negative, and are not NK cells. It may be helpful to > know that these samples had very poorly defined populations in the > forward vs. side scatter plot. I have not yet tried erythrocyte or > platelet stains to rule out aggregates or nucleated red blood > cells, nor have I tried staining isolated mononuclear cells. > > Does anyone have an idea what these cells may be? Could anither > cell fraction with these scatter qualities potentially be large > enough to make up over 50%? Has anyone had any experience with a > pathological state of which this may be symptomatic or for which > whole blood staining and lysis may yield erroneous results? Any > information or advice would be appreciated. Thank you in advance. > > Sincerely, > > Mikayla Kob > kob@cbr.med.harvard.edu > ===== `---------------------------------------------` | Maciej S. Simm | 525 E 68th Street | | Research Technician | Room N-805 | | Cornell Medical Center | Tel. 212.746.3428 | `---------------------------------------------` |www.cd4cd8.com | 917-734-6280 | AIM - XsimmX | `---------------------------------------------` __________________________________________________ Do You Yahoo!? Get personalized email addresses from Yahoo! Mail - only $35 a year! http://personal.mail.yahoo.com/
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