优化随机光学重建显微镜成像病理组织染色质结构

Q1 Health Professions

Current Protocols in Cytometry Pub Date : 2020-08-06 DOI:10.1002/cpcy.78

Jianquan Xu, Hongqiang Ma, Yang Liu

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引用次数: 2

摘要

在分子尺度上直接可视化高阶染色质结构对于理解染色质组织在许多生物过程中对基因表达的影响具有重要意义。了解病理过程中染色质结构的变化需要使用体内模型和临床样本，而福尔马林固定石蜡包埋(FFPE)组织是最广泛的保存形式。在这里，我们描述了PathSTORM的细节，这是一种优化的随机光学重建显微镜(STORM)方案，用于病理组织中密集排列的高阶染色质组织的高质量超分辨率成像。我们详细讨论了DNA和组蛋白荧光染色的方法，以及在病理组织样本中获得高质量STORM图像的关键技术因素。©2020 Wiley期刊llc基本协议1:病理组织中染色质的荧光染色基本协议2:STORM数据处理支持协议1:漂移校正支持协议2:图像重建支持协议3:苏木精;伊红(H&E)染色

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Optimized Stochastic Optical Reconstruction Microscopy for Imaging Chromatin Structure in Pathological Tissue

Direct visualization of higher‐order chromatin structure at the molecular scale is of great importance for understanding the impact of chromatin organization on gene expression in many biological processes. Understanding the changes in chromatin structure during pathological processes requires the use of in vivo models and clinical samples, and formalin‐fixed, paraffin‐embedded (FFPE) tissue is the most widespread form of preservation. Here we describe the details of PathSTORM, an optimized stochastic optical reconstruction microscopy (STORM) protocol for high‐quality super‐resolution imaging of densely packed higher‐order chromatin organization in pathological tissue. We discuss detailed methods for fluorescence staining of DNA and histone proteins, as well as the key technical factors for obtaining high‐quality STORM images in pathological tissue samples. © 2020 Wiley Periodicals LLC

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来源期刊

Current Protocols in Cytometry Health Professions-Medical Laboratory Technology

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期刊介绍： Published in affiliation with the International Society for Advancement of Cytometry, Current Protocols in Cytometry is a "best practices" collection that distills and organizes the absolute latest techniques from the top cytometry labs and specialists worldwide. It is the most complete set of peer-reviewed protocols for flow and image cytometry available.