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Rosetting, i.e. the capacity of red blood cells (iRBCs) infected with mature parasite stages to bind two or more uninfected red blood cells (RBCs) is a virulence factor of Plasmodium falciparum. This protocol describes an in vitro assay to monitor rosette formation by P. falciparum-infected red blood cells, including procedures for rosette enrichment, maintenance of rosetting phenotype and assays for rosetting with RBC labeled using lipophilic fluorescent probes.
Materials and Reagents
I. Human red blood cells and parasitized red blood cells
II. Culture medium reagents
III. Other materials
Use disposable professional gloves throughout.
This protocol was adapted from the following publications: Vigan-Womas et al. (2011) and Vigan-Womas et al. (2012). This work was supported by the Agence Nationale de la Recherche, contract ANR-07-MIME-021-0 (www.agence-nationale-recherche.fr/), and the 7th European Framework Program, FP7/2007-2013, (http://cordis.europa.eu/fp7/home_en.html) contract 242095, Evimalar.
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