期刊
CYTOMETRY PART A
卷 75A, 期 3, 页码 225-235出版社
WILEY
DOI: 10.1002/cyto.a.20647
关键词
malaria; flow cytometry; SYTO-16; bivariate analysis; Trucount (TM); P. falciparum; mice
资金
- Medicines for Malaria Venture (Geneva, Switzerland)
- Antonio Martinez
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 CD l mice infected with Plasmodium yoelii, Plasmodium vinckei, or Plasmodium chabaudi or from NODscid beta 2m-/- 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 (TM) 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. (c) 2008 International Society for Advancement of Cytometry
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