From: Brent Dorsett (brentd@nyct.net)
Date: Fri Dec 13 1996 - 13:59:10 EST
>Date: Thu, 12 Dec 1996 16:53:00 >To: Terry Hoy <Hoy@cardiff.ac.uk> >From: Brent Dorsett <brentd@nyct.net> >Subject: Re: PI staining of PBLs > >At 09:45 AM 12/12/96 GMT, you wrote: >> >>Mike Ormerod wrote : >>>I have run into a probelm staining peripheral blood cells >>>with propidium iodide for cell cycle analysis. ......... >> >>>We consitently obtain two peaks in the DNA histogram. A >>>lower peak from the lymphocytes and a peak from the >>>granulocytes with about 30% more fluorescence. >>> >>Mike, >>We noticed this several years ago looking at separated populations of >>lymphocytes, monocytes and granulocytes from peripheral blood >>(unpublished) and also with myeloid and erythroid cells separated from >>bone marrow. (Hodgetts et al J. Clin. Pathol. 1988; 41;1120-1124). >>The differences we noticed were all less than 10% - not as high as >>you are reporting. >>We thought this resulted from different degrees of chromatin >>condensation in the various lineages and opted for measuring >>separated populations each with its own 'lineage' control. >>All that work was by the Vindelov method. >>Terry. >> >> >>Dr. Terry Hoy >>Principal Research Officer >>Department of Haematology Telephone 44 1222 743458 >>University of Wales College of Medicine FAX 44 1222 744655 >>Heath Park E-mail hoy@cardiff.ac.uk >>CARDIFF CF4 4XN http://www.cf.ac.uk/uwcm/hg/hoy/index.html >>U.K. >> >> >Mike and Terry, >Several years ago while investigating patients with hypereosinophilic syndrome we found that bloods with high eosinophil counts that were stored for several hours before analysis with PI exhibited about 10% lower apparent DNA content. The same bloods examined immediately were normal. Eosinophils from normal blood seemed to show no such effect. Our unproven hypothesis was that in hypereosinophilic bloods storage ( RT ) induced an apoptotic effect. > >Brent Dorsett >Francesca Giancotti >Cancer Research Laboratory >Lenox Hill Hospital >NYC, NY >Tel 212-434-2476 >Fax 212-434-2497 >
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