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

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.

Keywords: DNA damage; DNA structure; chromatin; genomic integrity; super resolution microscopy.

Publication types

  • Review

MeSH terms

  • Animals
  • Chromatin / metabolism
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism*
  • DNA Breaks, Double-Stranded*
  • DNA Repair / physiology*
  • Fluorescent Dyes / chemistry
  • Histones / metabolism
  • Humans
  • Microscopy, Fluorescence / methods*
  • Molecular Probes / chemistry
  • Single Molecule Imaging / methods*

Substances

  • Chromatin
  • Fluorescent Dyes
  • Histones
  • Molecular Probes
  • DNA