Two-parameter single-molecule analysis for measurement of chromatin mobility

STAR Protoc. 2020 Dec 13;1(3):100223. doi: 10.1016/j.xpro.2020.100223. eCollection 2020 Dec 18.

Abstract

This protocol provides a two-parameter analysis of single-molecule tracking (SMT) trajectories of Halo-tagged histones in living adherent cell lines and unveils a chromatin mobility landscape composed of five chromatin types, ranging from low to high mobility. When the analysis is applied to Halo-tagged, chromatin-binding proteins, it associates chromatin interaction properties with known functions in a way that previously used SMT parameters did not. For complete information on the use and execution of this protocol, please refer to Lerner et al. (2020).

Keywords: Microscopy; Molecular biology; Single-molecule assays.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Chromatin / chemistry*
  • Chromatin / physiology
  • Chromosomes / metabolism
  • Fluorescent Antibody Technique / methods*
  • Histones / genetics
  • Humans
  • Protein Binding
  • Single Molecule Imaging / methods*

Substances

  • Chromatin
  • Histones