Journal of applied microbiology · 2001
Encystation of Acanthamoeba castellanii: dye uptake for assessment by flow cytometry and confocal laser scanning microscopy.
Сравнительное исследованиеСтатьяНазвание и аннотация: Перевод в очереди
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Connell C, Rutter A, Hill B, Suller M, Lloyd D
Аннотация
AIMS: To develop rapid means of distinguishing between cysts and trophozoites of the opportunistic pathogen, Acanthamoeba castellanii, the causative agent of keratitis.
METHODS AND RESULTS: Fluorescence of Congo Red, Calcoflor White was specific for the endocyst wall; trophozoites did not become fluorescent. The anionic oxonol dye, DiBAC4(3), did not penetrate the cytoplasmic membrane after short-term (<5 min) exposure, whereas cysts are permeable and become fluorescent. Confocal scanning laser microscopy confirmed these properties and large populations of organisms were analysed by flow cytometry.
CONCLUSION: These data provide a rapid alternative to traditional haemocytometer or plate counts for discrimination of trophozoites from cysts.
SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid and precise determination of the growth cycle of a dangerous ocular pathogen.
METHODS AND RESULTS: Fluorescence of Congo Red, Calcoflor White was specific for the endocyst wall; trophozoites did not become fluorescent. The anionic oxonol dye, DiBAC4(3), did not penetrate the cytoplasmic membrane after short-term (<5 min) exposure, whereas cysts are permeable and become fluorescent. Confocal scanning laser microscopy confirmed these properties and large populations of organisms were analysed by flow cytometry.
CONCLUSION: These data provide a rapid alternative to traditional haemocytometer or plate counts for discrimination of trophozoites from cysts.
SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid and precise determination of the growth cycle of a dangerous ocular pathogen.