I have seen that some groups use CSFE to stain some parasite species before infect cells to detect percetage of infected cells. Could I use CSFE to stain Trypanosoma cruzi and measure infection in cells? Someone have experience and advises?
fluorescent labels are maintained even after fixation for flow cytometry, in contrast to the case for green fluorescent protein (GFP) label, which diminishes after fixation . but the most uniform staining with CFSE labeling, and parasites can be detectable label up to 5 days (divisions) after labeling. Stationary-phase virulent promastigotes, attenuated L. chagasi promastigotes, L. donovani promastigotes, gene knockout L. donovani, and axenic L. chagasi amastigotes could also be labeled efficiently with CFSE . CFSE stain did not interfere with parasite motility or replication . The results indicate that Leishmania spp. can be short-term labeled with either CFSE or CM-DiI and detected by flow cytometry.
Recently, several immunological and molecular diagnostic tools have been developed for detection of Trypanosoma and Leishmania infections and their progressive quantification for severity , with Clinical Medical and Veterinary Medicine based Translational application.
Such as Real Time PCR. qPCR is fast, has a broad dynamic range and problems with cross-contamination is well reduced because there is no need to open reaction tubes for post-PCR analyses.
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