High-throughput detection of DNA double-strand breaks using image cytometry

Tyler L Fowler; Alison M Bailey; Bryan P Bednarz; Randall J Kimple

doi:10.2144/000114248

High-throughput detection of DNA double-strand breaks using image cytometry

Biotechniques. 2015 Jan 1;58(1):37-9. doi: 10.2144/000114248. eCollection 2015 Jan.

Authors

Tyler L Fowler¹, Alison M Bailey², Bryan P Bednarz³, Randall J Kimple⁴

Affiliations

¹ Departments of Medical Physics, School of Medicine and Public Health, University of Wisconsin, Madison, WI.
² Departments of Human Oncology, School of Medicine and Public Health, University of Wisconsin, Madison, WI.
³ Departments of Medical Physics, School of Medicine and Public Health, University of Wisconsin, Madison, WI; University of Wisconsin Carbone Comprehensive Cancer Center, School of Medicine and Public Health, University of Wisconsin, Madison, WI.
⁴ Departments of Human Oncology, School of Medicine and Public Health, University of Wisconsin, Madison, WI; University of Wisconsin Carbone Comprehensive Cancer Center, School of Medicine and Public Health, University of Wisconsin, Madison, WI.

Abstract

Assessment of γH2AX expression for studying DNA double-strand break formation is often performed by manual counting of foci using immunofluorescence microscopy, an approach that is laborious and subject to significant foci selection bias. Here we present a novel high-throughput method for detecting DNA double-strand breaks using automated image cytometry assessment of cell average γH2AX immunofluorescence. Our technique provides an expedient, high-throughput, objective, and cost-effective method for γH2AX analysis.

Keywords: DNA; double-strand breaks; image cytometry; γH2AX.

Publication types

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

MeSH terms

Cell Culture Techniques
DNA Breaks, Double-Stranded*
DNA Repair
HeLa Cells
Humans
Image Cytometry / methods*

Abstract

Publication types

MeSH terms

Grants and funding