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Encephalitozoon cuniculi : Quantitation of Parasites and Evaluation of Viability 1
Author(s) -
PETERSON NORMAN,
LIU JIUAN,
SHADDUCK JOHN
Publication year - 1988
Publication title -
the journal of protozoology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 0022-3921
DOI - 10.1111/j.1550-7408.1988.tb04123.x
Subject(s) - encephalitozoon cuniculi , propidium iodide , acridine orange , ethidium bromide , coulter counter , biology , stain , microbiology and biotechnology , chromatography , staining , chemistry , spore , microsporidia , biochemistry , dna , apoptosis , genetics , programmed cell death
Two methods (manual and automated) for quantitation of viable versus dead Encephalitozoon cuniculi are reported. The manual method uses ethidium bromide and acridine orange to stain dead and viable organisms, respectively. The stained organisms are visually differentiated with the aid of a fluorescence microscope. The automated method uses propidium iodide to stain dead parasites, which are differentiated from viable unstained parasites with the aid of a flow cytometer. An automated cell counter (Coulter Counter) was used to count rapidly large numbers of samples and to improve the sensitivity of counting low concentrations of parasites. These methods will enhance investigators' abilities to conduct quantitative experiments on host defense mechanisms against E. cuniculi.