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While experimenting with various permeablisation methods I noted that a sub
population of granulocytes did not shrink as much as the others.
At first I thought it might be an aged Neutrophil related phenomenon as can be
seen on old specimens run on a Coulter STKS.
Taking note of the percentages, however, showed that it was almost certainly the
Eos. that were resisting shrinkage.
These percentages matched the Eos in every sample tested and easily
distinguished them from Neuts using FS vs SS.
I did not follow this up any further, having no need as yet to look at Eos (and
besides, we don't have a sorter).
If I remember correctly, the "best" results were using very weak Nonidet P40 in
PBS pH 7.2.
0.05% for 5 minutes I think - without prior fixation.
I have a strong suspicion that Coulter and Cell-Dyn use a variation of this to
enhance Eosinophil separation.
As I said earlier, this may not be any good to you for any number of reasons,
but then again, it just might !
Wal Sharp
Sultan Qaboos University.
Sultanate of Oman.
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web