Encystation of Acanthamoeba castellanii: dye uptake for assessment by flow cytometry and confocal laser scanning microscopy

Connell, Christopher, Rutter, Andrew, Hill, B., Suller, M. and Lloyd, David ORCID: https://orcid.org/0000-0002-5656-0571 2001. Encystation of Acanthamoeba castellanii: dye uptake for assessment by flow cytometry and confocal laser scanning microscopy. Journal of Applied Microbiology 90 (5) , pp. 706-712. 10.1046/j.1365-2672.2001.01296.x

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Abstract

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. Conclusions: 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.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Schools > Biosciences
Publisher:	Wiley-Blackwell
ISSN:	1364-5072
Last Modified:	27 Oct 2022 08:34
URI:	https://orca.cardiff.ac.uk/id/eprint/62701

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