,Optimized protocol of RBCs lysis for immunophenotypic analysis in the peripheral blood of tree shre

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Tree shrew,an emerging experimental animal with its unique characters [1,2],attracts more and more attentions of scientists in studying animal models of human diseases,including infectious diseases [3],metabolic diseases [4],nerve diseases [5] and cancers [6].However,development and evaluation of animal models of tree shrews were halted due to lack of commercially available reagents and optimal protocols.Immunophenotyping the peripheral blood (PB) via flow cytometry is one of the fundamental analyses of host immune responses to pathogens or antigens,hematopoietic diseases,immune-related diseases,and stem cells in clinics and animal models.Pre-analytic lysis of red blood cells (RBCs) is considered to not only save antibodies and but also be critical for immunophenotypic analysis of the PB [7],although it is not necessary for some antibodies in clinical analysis.In the present study,the effectiveness of four reagents on RBCs lysis was examined and compared in the PB of tree shrew,followed by the optimization of protocols and conditions for the RBCs lysis,in order to obtain an optimal condition that gives effective lysis of RBCs and mostly viable cells for immunophenotypic analysis by flow cytometry.
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