Badri Ti, Eguren-Santamaria Iñaki, Fernandez-Pierola Eva, Sanmamed Miguel F
Immunobiology Department, Yale School of Medicine, New Haven, CT, United States.
Immunology and Immunotherapy Program CIMA, Universidad de Navarra, Pamplona, Spain.
Methods Cell Biol. 2023;174:31-41. doi: 10.1016/bs.mcb.2022.10.006. Epub 2022 Dec 14.
The human tumor microenvironment requires use of high-dimensional single-cell tools to uncover its cellular complexity and functional variety. For decades, flow cytometry has been the technology of choice to explore immune cell diversity in different pathological contexts. Recently, a new format for flow cytometry - termed mass cytometry - has been developed. It allows for simultaneous interrogation of more than 40 different molecular markers, without the need for spectral compensation, which significantly augments the ability of cytometry to evaluate complex cellular systems and processes. Currently, different multiparametric single-cell analysis approaches are being widely adopted to interrogate the human tumor microenvironment. However, important challenges must be addressed when solid tissues are analyzed by single-cell technologies. This protocol describes the challenge and better use of single-cell mass cytometry to dissect tumor infiltrating leukocytes from surgically resected tumoral lung tissues.
人类肿瘤微环境需要使用高维单细胞工具来揭示其细胞复杂性和功能多样性。几十年来,流式细胞术一直是探索不同病理背景下免疫细胞多样性的首选技术。最近,一种新的流式细胞术形式——质谱流式细胞术——已经被开发出来。它允许同时检测40多种不同的分子标记,而无需光谱补偿,这显著增强了流式细胞术评估复杂细胞系统和过程的能力。目前,不同的多参数单细胞分析方法正被广泛用于研究人类肿瘤微环境。然而,当通过单细胞技术分析实体组织时,必须应对一些重要挑战。本方案描述了使用单细胞质谱流式细胞术剖析手术切除的肿瘤肺组织中肿瘤浸润白细胞的挑战及更好的应用方法。