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流式细胞术分析血小板的免疫表型。

Immunophenotypic Analysis of Platelets by Flow Cytometry.

机构信息

Center for Platelet Research Studies, Dana-Farber/Boston Children's Cancer and Blood Disorders Center, Harvard Medical School, Boston, Massachusetts.

School of Biomedical Sciences, University of Western Australia, Perth, Australia.

出版信息

Curr Protoc. 2021 Jun;1(6):e178. doi: 10.1002/cpz1.178.

Abstract

Platelets are small but very abundant blood cells that play a key role in hemostasis, contributing to thrombus formation at sites of injury. The ability of platelets to perform this function, as well as functions in immunity and inflammation, is dependent on the presence of cell surface glycoproteins and changes in their quantity and conformation after platelet stimulation. In this article, we describe the characterization of platelet surface markers and platelet function using platelet-specific fluorescent probes and flow cytometry. Unlike traditional platelet tests, immunophenotypic analysis of platelets by flow cytometry allows the analysis of platelet function in samples with very low platelet counts as often encountered in clinical situations. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Immunophenotyping of platelet surface receptors Alternate Protocol: Fix-first method for immunophenotyping of platelet surface receptors Basic Protocol 2: Determination of platelet activation using P-selectin expression and/or PAC1 binding Basic Protocol 3: Determination of procoagulant platelets using annexin V binding or antibodies specific for coagulation factor V/Va or X/Xa Support Protocol: Preparation of isolated platelets.

摘要

血小板是体积较小但非常丰富的血细胞,在止血中发挥关键作用,有助于在损伤部位形成血栓。血小板发挥这种功能的能力,以及在免疫和炎症中的功能,依赖于细胞表面糖蛋白的存在,以及血小板刺激后其数量和构象的变化。在本文中,我们描述了使用血小板特异性荧光探针和流式细胞术来表征血小板表面标志物和血小板功能。与传统的血小板检测不同,流式细胞术对血小板的免疫表型分析允许在临床情况下经常遇到的血小板计数非常低的样本中分析血小板功能。© 2021 年 Wiley 期刊 LLC。 基本方案 1:血小板表面受体的免疫表型分析 可选方案:用于血小板表面受体免疫表型分析的固定优先法 基本方案 2:使用 P-选择素表达和/或 PAC1 结合测定血小板活化 基本方案 3:使用 Annexin V 结合或针对凝血因子 V/Va 或 X/Xa 的抗体测定促凝血小板 支持方案:分离血小板的制备。

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