Re: PI staining of PBLs
Brent Dorsett (brentd@nyct.net)
Fri, 13 Dec 1996 13:59:10 -0500 (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(unpublished) and also with myeloid and erythroid cells separated from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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