Haddad Tariq Sami, Friedl Peter, Farahani Navid, Treanor Darren, Zlobec Inti, Nagtegaal Iris
Department of Pathology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands.
Department of Cell Biology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands.
Nat Protoc. 2021 Nov;16(11):4945-4962. doi: 10.1038/s41596-021-00611-4. Epub 2021 Oct 29.
Analysis of three-dimensional patient specimens is gaining increasing relevance for understanding the principles of tissue structure as well as the biology and mechanisms underlying disease. New technologies are improving our ability to visualize large volume of tissues with subcellular resolution. One resource often overlooked is archival tissue maintained for decades in hospitals and research archives around the world. Accessing the wealth of information stored within these samples requires the use of appropriate methods. This tutorial introduces the range of sample preparation and microscopy approaches available for three-dimensional visualization of archival tissue. We summarize key aspects of the relevant techniques and common issues encountered when using archival tissue, including registration and antibody penetration. We also discuss analysis pipelines required to process, visualize and analyze the data and criteria to guide decision-making. The methods outlined in this tutorial provide an important and sustainable avenue for validating three-dimensional tissue organization and mechanisms of disease.
对三维患者标本进行分析对于理解组织结构原理以及疾病背后的生物学和机制越来越重要。新技术正在提高我们以亚细胞分辨率可视化大量组织的能力。一个经常被忽视的资源是在世界各地医院和研究档案中保存了数十年的存档组织。获取这些样本中存储的丰富信息需要使用适当的方法。本教程介绍了可用于存档组织三维可视化的样本制备和显微镜方法的范围。我们总结了相关技术的关键方面以及使用存档组织时遇到的常见问题，包括配准和抗体穿透。我们还讨论了处理、可视化和分析数据所需的分析流程以及指导决策的标准。本教程中概述的方法为验证三维组织结构和疾病机制提供了一条重要且可持续的途径。
