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Plasmodium berghei-infected red cells sorted according to DNA content

Published online by Cambridge University Press:  06 April 2009

R. J. Howard  and
F. L. Battye
R. J. Howard
Affiliation:
Laboratory of Immunoparasitology, The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria 3050, Australia
F. L. Battye
Affiliation:
Laboratory of Immunoparasitology, The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria 3050, Australia
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Summary

A cell-sorting method is described for the analysis and separation of red blood cells in Plasmodium berghei-infected mouse blood based on their DNA content. This method involves a selective uptake of the bis-benzimidazole dye 33258 Hoechst, a DNA-binding dye, by red blood cells containing parasites. Infected blood is incubated at 37 °C with the dye then washed at 4 °C to remove unbound dye. Uninfected cells are then non-fluorescent at the characteristic wavelengths for 33258 Hoechst excitation and emission, whereas parasitized cells display fluorescence intensities in approximately direct proportion to the number of parasite nuclei (i.e. amount of parasite DNA) within the cell and can be sorted accordingly. Providing cells were incubated in a complex nutrient medium during dye uptake at 37°C, the sorted parasite cells produced lethal P. berghei infections when injected into BALB/c mice. The dyelabelling technique is simple and sufficient red blood cells at various stages of infection can be collected for biochemical or immunochemical studies by cell sorting.

Type
Research Article
Information
Parasitology , Volume 78 , Issue 3 , June 1979 , pp. 263 - 270
Copyright
Copyright © Cambridge University Press 1979

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References

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