Greetings flowers, 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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