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利用超分辨率荧光显微镜理解DNA的组织、损伤及修复

Understanding DNA organization, damage, and repair with super-resolution fluorescence microscopy.

作者信息

Miriklis Esther L, Rozario Ashley M, Rothenberg Eli, Bell Toby D M, Whelan Donna R

机构信息

School of Chemistry, Monash University, Clayton, VIC, Australia.

Department of Biochemistry and Molecular Pharmacology, Perlmutter Cancer Center, New York University School of Medicine, New York, NY, United States of America.

出版信息

Methods Appl Fluoresc. 2021 May 5;9(3). doi: 10.1088/2050-6120/abf239.

Abstract

Super-resolution microscopy (SRM) comprises a suite of techniques well-suited to probing the nanoscale landscape of genomic function and dysfunction. Offering the specificity and sensitivity that has made conventional fluorescence microscopy a cornerstone technique of biological research, SRM allows for spatial resolutions as good as 10 nanometers. Moreover, single molecule localization microscopies (SMLMs) enable examination of individual molecular targets and nanofoci allowing for the characterization of subpopulations within a single cell. This review describes how key advances in both SRM techniques and sample preparation have enabled unprecedented insights into DNA structure and function, and highlights many of these new discoveries. Ongoing development and application of these novel, highly interdisciplinary SRM assays will continue to expand the toolbox available for research into the nanoscale genomic landscape.

摘要

超分辨率显微镜(SRM)包含一系列非常适合探究基因组功能与功能障碍纳米尺度状况的技术。SRM具备使传统荧光显微镜成为生物学研究基石技术的特异性和灵敏度,能够实现低至10纳米的空间分辨率。此外,单分子定位显微镜(SMLM)能够检测单个分子靶点和纳米焦点,从而对单个细胞内的亚群进行表征。本综述描述了SRM技术和样品制备方面的关键进展如何实现了对DNA结构和功能前所未有的深入了解,并突出了其中许多新发现。这些新颖的、高度跨学科的SRM检测方法的持续开发和应用将继续扩充可用于纳米尺度基因组研究的工具库。

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