For decades, histopathology with routine hematoxylin and eosin staining has been and remains the gold standard for reaching a morphologic diagnosis in tissue samples from humans and veterinary species. However, within the past decade, there has been exponential growth in advanced techniques for in situ tissue biomarker imaging that bridge the divide between anatomic and molecular pathology. It is now possible to simultaneously observe localization and expression magnitude of multiple protein, nucleic acid, and molecular targets in tissue sections and apply machine learning to synthesize vast, image-derived datasets. As these technologies become more sophisticated and widely available, a team-science approach involving subspecialists with medical, engineering, and physics backgrounds is critical to upholding quality and validity in studies generating these data. The purpose of this manuscript is to detail the scientific premise, tools and training, quality control, and data collection and analysis considerations needed for the most prominent advanced imaging technologies currently applied in tissue sections: immunofluorescence, in situ hybridization, laser capture microdissection, matrix-assisted laser desorption ionization imaging mass spectrometry, and spectroscopic/optical methods. We conclude with a brief overview of future directions for ex vivo and in vivo imaging techniques.

中文翻译：

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超越 H&E：原位组织生物标志物成像的先进技术。

几十年来，常规苏木精和伊红染色的组织病理学一直是并且仍然是对人类和兽医物种的组织样本进行形态学诊断的金标准。然而，在过去的十年中，原位组织生物标志物成像的先进技术呈指数级增长，弥合了解剖学和分子病理学之间的鸿沟。现在可以同时观察组织切片中多个蛋白质、核酸和分子靶标的定位和表达量，并应用机器学习来合成大量的图像衍生数据集。随着这些技术变得更加复杂和广泛应用，涉及具有医学、工程和物理背景的亚专家的团队科学方法对于维护生成这些数据的研究的质量和有效性至关重要。本手稿的目的是详细介绍目前应用于组织切片的最突出的先进成像技术所需的科学前提、工具和培训、质量控制以及数据收集和分析注意事项：免疫荧光、原位杂交、激光捕获显微切割、矩阵-辅助激光解吸电离成像质谱法和光谱/光学方法。最后，我们简要概述了离体和体内成像技术的未来方向。