Protocol to study immunodynamics in the tumor microenvironment using a tyramide signal amplification-based immunofluorescent multiplex panel

利用基于酪胺信号放大的免疫荧光多重检测方法研究肿瘤微环境免疫动力学的方案

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作者:Jane Siu-Fan Li ,Philip Chiu-Tsun Tang ,Chun Kit K Choi ,Alex Siu-Wing Chan ,Calvin Sze-Hang Ng ,Ka-Fai To ,Patrick Ming-Kuen Tang

Abstract

Immunodynamics in the tumor microenvironment can be precisely examined by using multiple antigen identification approaches. Here, we present a protocol for capturing expression levels of multiple target proteins in the same specimen at single-cell resolution using a tyramide signal amplification-based immunofluorescent multiplexing system. We describe steps for tumor tissue microarray preparation, multiplex immunohistochemistry staining, image acquisition, and quantification. This protocol can quantify immune cells in tissues from patients or experimental disease models at a protein level. For complete details on the use and execution of this protocol, please refer to Chung et al. (2023),1 Tang et al. (2022),2 and Tang et al. (2022).3. Keywords: Cancer; Clinical Protocol; Immunology; Microscopy.

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