Observation of intracellular protozoan development by tissue culture on cover glasses and detection by fresh observation and staining methods

Abstract

Isospora belli, Cryptosporidium parvum sporozoites and Microsporidial spores were infected into culture cells. The developmental stages of these intracellular protozoa were described after the fresh observation and staining of infected cells on cover glasses. Paraffin tissue section was performed and compared to cover glass staining. Fresh observation was more advantages for the observation of the movement of I. belli merozoites but the development of merogony of Cryptosporidium could not be notified by fresh observation. The merogony stages of I. belli from paraffin tissue sections were not clear, compared to cover glass staining but the trophozoites of Cryptosporidium were clearly demonstrated by paraffin tissue sections. The typical characters of belt-like stripes of Microsporidial spores from culture were clearly observed from cover glass staining. In conclusion, cover glass staining could be used for an alternative method for the detection of intracellular protozoa from cultures. However, the properstaining methodsshould be selected for each protozoan parasite.