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
