Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Institute for Immunology and Center for AIDS Research, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
STAR Protoc. 2020 Oct 29;1(3):100154. doi: 10.1016/j.xpro.2020.100154. eCollection 2020 Dec 18.
Recent advances in high-resolution multiparametric flow cytometry enable ever deeper analysis of human lymphocyte subsets that require rigorous methodology development and optimization. Here, we detail methods to characterize glycosylated Sialyl-Lewis (SLe)- or cutaneous lymphocyte-associated antigen (CLA)-expressing CD4+ T cells using two separate multiparametric flow cytometry panels enabling the identification of memory subsets, Th subsets, and expression of diverse activation markers and chemokine receptors. The proposed protocol allows optimal resolution of the measured parameters while minimizing background in a 25-parameter experiment. For complete details on the use and execution of this protocol, please refer to Colomb et al. (2020).
近年来，高分辨率多参数流式细胞术的进展使得对人类淋巴细胞亚群的分析更加深入，这需要严格的方法学开发和优化。在这里，我们详细介绍了使用两个独立的多参数流式细胞术面板来描述表达糖基化 Sialyl-Lewis（SLe）或皮肤淋巴细胞相关抗原（CLA）的 CD4+T 细胞的方法，这两个面板可用于识别记忆亚群、Th 亚群以及多种活化标志物和趋化因子受体的表达。该方案在 25 参数实验中最大限度地提高了测量参数的分辨率，同时最小化了背景。有关此方案的使用和执行的完整详细信息，请参阅 Colomb 等人（2020 年）。
