Document Detail

Quantitative measurement of Plasmodium-infected erythrocytes in murine models of malaria by flow cytometry using bidimensional assessment of SYTO-16 fluorescence.
MedLine Citation:
PMID:  18785271     Owner:  NLM     Status:  MEDLINE    
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 (> or =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(scidbeta2m-/-) 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.
María Belén Jiménez-Díaz; Teresa Mulet; Vanesa Gómez; Sara Viera; Angela Alvarez; Helena Garuti; Yolanda Vázquez; Alejandra Fernández; Javier Ibáñez; Magdalena Jiménez; Domingo Gargallo-Viola; Iñigo Angulo-Barturen
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytometry. Part A : the journal of the International Society for Analytical Cytology     Volume:  75     ISSN:  1552-4930     ISO Abbreviation:  Cytometry A     Publication Date:  2009 Mar 
Date Detail:
Created Date:  2009-02-19     Completed Date:  2009-06-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101235694     Medline TA:  Cytometry A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  225-35     Citation Subset:  IM    
Copyright Information:
2008 International Society for Advancement of Cytometry.
GlaxoSmithKline R & D, ID CEDD Diseases of the Developing World, Tres Cantos, Spain.
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MeSH Terms
Disease Models, Animal
Erythrocytes / cytology,  parasitology*
Flow Cytometry / methods*
Fluorescent Dyes / chemistry*,  metabolism
Malaria / parasitology*
Mice, Inbred Strains
Mice, SCID
Plasmodium / isolation & purification*
Reg. No./Substance:
0/Fluorescent Dyes

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