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Separation of Isospora (Toxoplasma) gondii cysts and cystozoites from mouse brain tissue by continuous density - gradient centrifugation

Published online by Cambridge University Press:  06 April 2009

A. W. C. A. Cornelissen
Affiliation:
Department of Tropical Veterinary Medicine and Protozoology, State University Utrecht, Biltstraat 172, 3572 BP Utrecht, The Netherlands
J. P. Overdulve
Affiliation:
Department of Tropical Veterinary Medicine and Protozoology, State University Utrecht, Biltstraat 172, 3572 BP Utrecht, The Netherlands
J. M. Hoenderboom
Affiliation:
Department of Tropical Veterinary Medicine and Protozoology, State University Utrecht, Biltstraat 172, 3572 BP Utrecht, The Netherlands

Summary

A simple, quick and reproducible method consisting of density-gradient centrifugation of homogenized infected mouse brain tissue on Percoll is described for the isolation and purification of cysts of Isospora (Toxoplasma) gondii. A 100% recovery of cysts, with 74·2% in a single fraction with a specific gravity of 1·056, was obtained by overlaying homogenates of infected mouse brains on a pre-formed Percoll gradient and centrifugation at low g forces. With this procedure recovery was independent of the age of the cysts. Titration of purified cystozoites showed there to be no loss of infectivity.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1981

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