Cells and Stripes: A novel quantitative photo-manipulation technique

0473046 - ÚMG 2017 RIV GB eng J - Journal Article
Mistrik, M. - Veselá, E. - Fürst, T. - Hanzlíková, Hana - Frydrych, I. - Gursky, J. - Majera, D. - Bártek, Jiří
Cells and Stripes: A novel quantitative photo-manipulation technique.
Scientific Reports. Roč. 6, JAN 18 (2016), č. článku 19567. ISSN 2045-2322. E-ISSN 2045-2322
R&D Projects: GA ČR GA13-17555S; GA MŠMT(CZ) LO1304; GA MŠMT(CZ) ED2.3.30.0004; GA MŠMT(CZ) ED2.3.30.0041
Institutional support: RVO:68378050
Keywords : double-strand breaks * dna-damage-response * fanconi anemia/brca pathway * repair * mdc1 * chromosomes * inhibition * machinery * retention * proteins
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 4.259, year: 2016

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.
Permanent Link: http://hdl.handle.net/11104/0270214