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Cryptosporidium parvum

Kurt J. Huebner huebner at earth.execpc.com
Wed Nov 1 21:45:27 EST 1995


Sir. I am a graduate student researching the interaction of the C. parvum
sporozoite with CACO-2 and murine macrophage cell lines (with and without
chemotherapeutics).  The excystation process I use is the standard Na
taurocholate and trypsin methodology.  I am getting great sporozoite
return.  However, my problem appears to be maintaining their viablility (I
have made the assumption based upon phase microcroscopy, that excysting
sporozoites are viable).  I am presently infecting my cells lines within
less than one-half hour of excystation.  Infection is discernable, but the
rates have been very low. I am hoping that one of you accomplished
gentlemen will be willing to review my procedure with me.  If you have a
method that has worked well for you I would hope you would share your
experience. 
  In addition, aside from acid-fast stains, auramine-rhodamin and Geimsa
are their other stains that will better demonstrate the intracellular
sporozoite to merozoite stages?  I have also worked with propidium iodide/FDA
and acridine orange. I don't have access to anti-sporozoite antibodies, so
IDFA is not an option. 
  I realize that I am asking for an extensive dialogue.  I thank you in
advance for your consideration of my request and for any information you
may be able to provide.  This is my first attempt at posting, so if I have
errored in some way ...

Kurt J Huebner
huebner at execpc.com





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