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Quantitative measurement of Plasmodium ‐infected erythrocytes in murine models of malaria by flow cytometry using bidimensional assessment of SYTO‐16 fluorescence
Author(s) -
JiménezDíaz María Belén,
Mulet Teresa,
Gómez Vanesa,
Viera Sara,
Alvarez Angela,
Garuti Helena,
Vázquez Yolanda,
Fernández Alejandra,
Ibáñez Javier,
Jiménez Magdalena,
GargalloViola Domingo,
AnguloBarturen Iñigo
Publication year - 2009
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20647
Subject(s) - plasmodium falciparum , flow cytometry , plasmodium yoelii , plasmodium chabaudi , biology , malaria , cytometry , microbiology and biotechnology , apicomplexa , virology , plasmodium (life cycle) , parasite hosting , parasitemia , immunology , world wide web , computer science
Flow cytometry is a powerful tool for measuring parasitemias in murine malaria models used to test new antimalarials. Measurement of the emission of the nonpermeable nucleic acid dye YOYO‐1 (at 530 and 585 nm after excitation at 488 nm) allowed the unambiguous detection of low parasitemias (≥0.01%) but required prolonged fixation and permeabilization of the sample. Thus, we tested whether this issue could be overcome by use of the cell‐permeant dye SYTO‐16 with this same bidimensional method. Blood samples from CD1 mice infected with Plasmodium yoelii , Plasmodium vinckei , or Plasmodium chabaudi or from NOD scidβ2m‐/‐ engrafted with human erythrocytes and infected with P. falciparum were stained with SYTO‐16 in the presence or absence of TER‐119 mAb (for engrafted mice) in 96‐well plate format and acquired in Trucount™ tubes. Bidimensional analysis with SYTO‐16 was quantitatively equivalent to YOYO‐1. Moreover, by combining SYTO‐16 with the use of TER‐119‐PE antimouse erythrocyte mAb and Trucount tubes, the measurement of the concentration of P. falciparum ‐infected erythrocytes over a range of five orders of magnitude was achieved. Bidimensional analysis using SYTO‐16 can be used to accurately measure the concentration of Plasmodium spp.‐infected erythrocytes in mice without complex sample preparation. © 2008 International Society for Advancement of Cytometry