A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development

Microscopic examination of Giemsa-stained thin blood smears remains the gold standard method used to quantify and stage malaria parasites. However, this technique is tedious, and requires trained microscopists. We have developed a fast and simple flow cytometry method to quantify and stage, various...

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Main Authors:	Malleret, Benoît, Claser, Carla, Ong, Alice Soh Meoy, Suwanarusk, Rossarin, Sriprawat, Kanlaya, Howland, Shanshan Wu, Russell, Bruce, Nosten, Francois, Rénia, Laurent
Format:	Online
Language:	English
Published:	Nature Publishing Group 2011
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3216599/

id	pubmed-3216599
recordtype	oai_dc
spelling	pubmed-32165992011-12-22 A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development Malleret, Benoît Claser, Carla Ong, Alice Soh Meoy Suwanarusk, Rossarin Sriprawat, Kanlaya Howland, Shanshan Wu Russell, Bruce Nosten, Francois Rénia, Laurent Article Microscopic examination of Giemsa-stained thin blood smears remains the gold standard method used to quantify and stage malaria parasites. However, this technique is tedious, and requires trained microscopists. We have developed a fast and simple flow cytometry method to quantify and stage, various malaria parasites in red blood cells in whole blood or in vitro cultured Plasmodium falciparum. The parasites were stained with dihydroethidium and Hoechst 33342 or SYBR Green I and leukocytes were identified with an antibody against CD45. Depending on the DNA stains used, samples were analyzed using different models of flow cytometers. This protocol, which does not require any washing steps, allows infected red blood cells to be distinguished from leukocytes, as well as allowing non-infected reticulocytes and normocytes to be identified. It also allows assessing the proportion of parasites at different developmental stages. Lastly, we demonstrate how this technique can be applied to antimalarial drug testing. Nature Publishing Group 2011-10-14 /pmc/articles/PMC3216599/ /pubmed/22355635 http://dx.doi.org/10.1038/srep00118 Text en Copyright © 2011, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-sa/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-ShareALike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Malleret, Benoît Claser, Carla Ong, Alice Soh Meoy Suwanarusk, Rossarin Sriprawat, Kanlaya Howland, Shanshan Wu Russell, Bruce Nosten, Francois Rénia, Laurent
spellingShingle	Malleret, Benoît Claser, Carla Ong, Alice Soh Meoy Suwanarusk, Rossarin Sriprawat, Kanlaya Howland, Shanshan Wu Russell, Bruce Nosten, Francois Rénia, Laurent A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development
author_facet	Malleret, Benoît Claser, Carla Ong, Alice Soh Meoy Suwanarusk, Rossarin Sriprawat, Kanlaya Howland, Shanshan Wu Russell, Bruce Nosten, Francois Rénia, Laurent
author_sort	Malleret, Benoît
title	A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development
title_short	A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development
title_full	A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development
title_fullStr	A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development
title_full_unstemmed	A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development
title_sort	rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development
description	Microscopic examination of Giemsa-stained thin blood smears remains the gold standard method used to quantify and stage malaria parasites. However, this technique is tedious, and requires trained microscopists. We have developed a fast and simple flow cytometry method to quantify and stage, various malaria parasites in red blood cells in whole blood or in vitro cultured Plasmodium falciparum. The parasites were stained with dihydroethidium and Hoechst 33342 or SYBR Green I and leukocytes were identified with an antibody against CD45. Depending on the DNA stains used, samples were analyzed using different models of flow cytometers. This protocol, which does not require any washing steps, allows infected red blood cells to be distinguished from leukocytes, as well as allowing non-infected reticulocytes and normocytes to be identified. It also allows assessing the proportion of parasites at different developmental stages. Lastly, we demonstrate how this technique can be applied to antimalarial drug testing.
publisher	Nature Publishing Group
publishDate	2011
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3216599/
_version_	1611487905461567488

A rapid and robust tri-color flow cytometry assay for monitoring malaria parasite development

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