Wickenhauser Claudia, Bethmann Daniel, Feng Zipei, Jensen Shawn M, Ballesteros-Merino Carmen, Massa Chiara, Steven Andre, Bauer Marcus, Kaatzsch Peter, Pazaitis Nikolaos, Toma Georgiana, Bifulco Carlo B, Fox Bernard A, Seliger Barbara
Medical Faculty, Institute of Pathology, Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.
Robert W. Franz Cancer Center, Earle A. Chiles Research Institute, Providence Portland Medical Center, Portland, OR, USA.
Methods Mol Biol. 2019;1913:13-31. doi: 10.1007/978-1-4939-8979-9_2.
Histomorphology has significantly changed over the last decades due to technological achievements in immunohistochemistry (IHC) for the visualization of specific proteins and in molecular pathology, particularly in the field of in situ hybridization of small oligonucleotides and amplification of DNA and RNA amplicons. With an increased availability of suitable methods, the demands regarding the observer of histomorphological slides were the supply of complex quantitative data as well as more information about protein expression and cell-cell interactions in tissue sections. Advances in fluorescence-based multiplexed IHC techniques, such as multispectral imaging (MSI), allow the quantification of multiple proteins at the same tissue section. In histopathology, it is a well-known technique for over a decade yet harboring serious problems concerning quantitative preciseness and tissue autofluorescence of multicolor staining when using formalin-fixed, paraffin-embedded (FFPE) tissue specimen. In recent years, milestones in tissue preparation, fluorescent dyes, hardware imaging, and software analysis were achieved including automated tissue segmentation (e.g., tumor vs. stroma) as well as in cellular and subcellular multiparameter analysis.This chapter covers the role that MSI plays in anatomic pathology for the analysis of FFPE tissue sections, discusses the technical aspects of MSI, and provides a review of its application in the characterization of immune cell infiltrates and beyond regarding its prognostic and predictive value and its use for guidance of clinical decisions for immunotherapeutic strategies.
在过去几十年中，由于免疫组织化学（IHC）在特定蛋白质可视化方面以及分子病理学领域，特别是在小寡核苷酸原位杂交和DNA及RNA扩增子扩增方面的技术成就，组织形态学发生了显著变化。随着合适方法的日益增多，对组织形态学切片观察者的要求是提供复杂的定量数据以及更多关于组织切片中蛋白质表达和细胞间相互作用的信息。基于荧光的多重免疫组化技术的进展，如多光谱成像（MSI），使得在同一组织切片上对多种蛋白质进行定量成为可能。在组织病理学中，这是一项已为人熟知十多年的技术，但在使用福尔马林固定、石蜡包埋（FFPE）组织标本时，在多色染色的定量准确性和组织自发荧光方面仍存在严重问题。近年来，在组织制备、荧光染料、硬件成像和软件分析方面取得了里程碑式的进展，包括自动组织分割（如肿瘤与基质）以及细胞和亚细胞多参数分析。本章涵盖了MSI在解剖病理学中对FFPE组织切片分析所起的作用，讨论了MSI的技术方面，并综述了其在免疫细胞浸润特征分析及其他方面的应用，包括其预后和预测价值以及在免疫治疗策略临床决策指导中的应用。
