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With all of the discussion on DNA staining going on, I have a sort of
similar question.
I have been running samples for an investigator who is using Hoechst (33258
and 33342) staining to identify malaria parasitised red blood cells. So
far the different stages of parasite development have been readily
identifiable, BUT there is one little observation that is bugging me:
She has two types of samples. Some of which are totally unparasitised, and
others of which a proportion of the RBCs are parasitised. The negative
RBCs in the parasitised samples seem to always have a lower fluorescence
than the unparasitised cells. Why should this be? and should I be
concerned?
The only explanation I can think of is that the stain is binding to the DNA
of the parasites and leaving a lower amount of unbound dye to contribute to
the background fluoresence of the unparasitised cells. Is this sensible,
or am I way off the mark?
Any thoughts appreciated.
Andrew Mitchell
School of Microbiology and Immunology
University of New South Wales
Australia
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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;
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