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 |
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 |
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Date Type: | Publication |
Status: | Published |
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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