In vivo assessment of rodent Plasmodium parasitemia and merozoite invasion by flow cytometry

Patrick M. Lelliott*, Brendan J. McMorran, Simon J. Foote, Gaetan Burgio

*Corresponding author for this work

    Research output: Contribution to journalArticle

    7 Citations (Scopus)

    Abstract

    During blood stage infection, malaria parasites invade, mature, and replicate within red blood cells (RBCs). This results in a regular growth cycle and an exponential increase in the proportion of malaria infected RBCs, known as parasitemia. We describe a flow cytometry based protocol which utilizes a combination of the DNA dye Hoechst, and the mitochondrial membrane potential dye, JC-1, to identify RBCs which contain parasites and therefore the parasitemia, of in vivo blood samples from Plasmodium chabaudi adami DS infected mice. Using this approach, in combination with fluorescently conjugated antibodies, parasitized RBCs can be distinguished from leukocytes, RBC progenitors, and RBCs containing Howell-Jolly bodies (HJ-RBCs), with a limit of detection of 0.007% parasitemia. Additionally, we outline a method for the comparative assessment of merozoite invasion into two different RBC populations. In this assay RBCs, labeled with two distinct compounds identifiable by flow cytometry, are transfused into infected mice. The relative rate of invasion into the two populations can then be assessed by flow cytometry based on the proportion of parasitized RBCs in each population over time. This combined approach allows the accurate measurement of both parasitemia and merozoite invasion in an in vivo model of malaria infection.

    Original languageEnglish
    Article numbere52736
    Pages (from-to)1-8
    Number of pages8
    JournalJournal of Visualized Experiments
    Volume2015
    Issue number98
    DOIs
    Publication statusPublished - 5 Apr 2015

    Keywords

    • Chabaudi
    • Flow cytometry
    • In vivo
    • Infection
    • Invasion
    • Issue 98
    • JC-1
    • Malaria
    • Merozoite
    • Parasitemia
    • Plasmodium

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