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There are two cell preparatory methods: one analyzes whole cells and
the other lyses the cells and analyzes the released nuclei.
Gating is critical to determine an accurate percentage apoptotic.
Each tube contains "live" cells, apoptotic cells, debris, and
clumps.
I use a whole cell method with a linear fluorescence scale and
use both LS gates [mainly to exclude debris] and FL gates [mainly
to exclude doublets and clumps].
Finally, you need to verify that this assay works for your system
by alternate methods [counting the apoptotic cells under the scope
is easiest and perhaps best].
is easiest and perhaps best].
is easiest and perhaps best].
is easiest and perhaps best].
Good luck,
Tom Mc Closkey
North Shore University Hospital
Long Island, NY
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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
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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