High-throughput Evaluation of Protein Migration and Localization after Laser Micro-Irradiation

Sci Rep. 2019 Feb 28;9(1):3148. doi: 10.1038/s41598-019-39760-8.

Abstract

DNA- and histone-related research frequently comprises the quantitative analysis of protein modifications, such as histone phosphorylation. Analysis of accumulation and disappearance of protein foci are used to monitor DNA damage and repair kinetics. If the protein of interest doesn't accumulate in foci, laser micro-irradiation of single nuclei provides an alternative method to monitor DNA repair proteins and histone dynamics at the DNA damage site. We have developed an automated evaluation tool for standardized, high-throughput analysis of micro-irradiated cells featuring single cell background subtraction and detection across multiple fluorescence channels, allowing for robust statistics.

Publication types

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

MeSH terms

  • DNA Breaks, Double-Stranded / radiation effects
  • DNA Damage / genetics
  • DNA Damage / radiation effects
  • DNA Repair / genetics
  • DNA Repair / radiation effects
  • Histone Code / genetics*
  • Histones / chemistry
  • Histones / genetics*
  • Humans
  • Lasers / adverse effects
  • Microscopy, Fluorescence
  • Phosphorylation / radiation effects
  • Protein Processing, Post-Translational / genetics*
  • Proteins / chemistry
  • Proteins / genetics*
  • Single-Cell Analysis

Substances

  • Histones
  • Proteins