Quantitative Measurement of Plasmodium-Infected Erythrocytes in Murine Models of Malaria by Flow Cytometry Using Bidimensional Assessment of SYTO-16 Fluorescence
Quantitative Measurement of Plasmodium-Infected Erythrocytes in Murine Models of Malaria by Flow Cytometry Using Bidimensional Assessment of SYTO-16 Fluorescence
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DOI：
10.1002/cyto.a.20647
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发表时间：
2009-03-01
期刊：
影响因子：
3.7
通讯作者：
Angulo-Barturen, Inigo
中科院分区：
文献类型：
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作者：
Jimenez-Diaz, Maria Belen;Mulet, Teresa;Angulo-Barturen, Inigo
流式细胞术是一个强大的工具，用于测量寄生虫血症的鼠疟疾模型用于测试新的抗疟药。测量非渗透性核酸染料YOYO-1的发射（在488 nm激发后在530和585 nm处）允许明确检测低寄生虫血症（>= 0.01%），但需要延长样品的固定和透化。因此，我们测试了这个问题是否可以通过使用细胞渗透染料SYTO-16与这种相同的二维方法来克服。在TER-119 mAb存在或不存在的情况下（对于移植的小鼠），在96孔板形式中用SYTO-16对来自感染约氏疟原虫、温氏疟原虫或夏氏疟原虫的CD 1小鼠或来自移植有人红细胞并感染恶性疟原虫的NODscid β 2 m-/-的血液样品进行染色，并在Trucount（TM）管中获得。用SYTO-16进行的二维分析在定量上与YOYO-1相当。此外，通过将SYTO-16与TER-119-PE抗小鼠红细胞mAb和Trucount管的使用相结合，实现了在五个数量级范围内测量恶性疟原虫感染的红细胞的浓度。使用SYTO-16的二维分析可用于准确测量疟原虫属的浓度。感染红细胞的小鼠中，而无需复杂的样品制备。(c)2008年国际细胞计数促进学会
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