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Sci Rep. 2016 Jan 18;6:19567. doi: 10.1038/srep19567.

Cells and Stripes: A novel quantitative photo-manipulation technique.

Author information

1
Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.
2
Institute of Molecular Genetics of the ASCR, v. v. i., Prague, Czech Republic.
3
Danish Cancer Society Research Center, Copenhagen, Denmark.

Abstract

Laser micro-irradiation is a technology widely used in the DNA damage response, checkpoint signaling, chromatin remodeling and related research fields, to assess chromatin modifications and recruitment of diverse DNA damage sensors, mediators and repair proteins to sites of DNA lesions. While this approach has aided numerous discoveries related to cell biology, maintenance of genome integrity, aging and cancer, it has so far been limited by a tedious manual definition of laser-irradiated subcellular regions, with the ensuing restriction to only a small number of cells treated and analyzed in a single experiment. Here, we present an improved and versatile alternative to the micro-irradiation approach: Quantitative analysis of photo-manipulated samples using innovative settings of standard laser-scanning microscopes. Up to 200 cells are simultaneously exposed to a laser beam in a defined pattern of collinear rays. The induced striation pattern is then automatically evaluated by a simple algorithm, which provides a quantitative assessment of various laser-induced phenotypes in live or fixed cells. Overall, this new approach represents a more robust alternative to existing techniques, and provides a versatile tool for a wide range of applications in biomedicine.

PMID:
26777522
PMCID:
PMC4726120
DOI:
10.1038/srep19567
[Indexed for MEDLINE]
Free PMC Article

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