ŠIMEČKOVÁ, Šárka, Radek FEDR, Ján REMŠÍK, Z. KAHOUNOVÁ, Eva SLABÁKOVÁ and Karel SOUČEK. Multiparameter cytometric analysis of complex cellular response. Cytometry Part A. HOBOKEN: John Wiley & Sons, 2018, 93A, No 2, p. 239-248. ISSN 1552-4922. Available from: https://dx.doi.org/10.1002/cyto.a.23295.
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Basic information
Original name Multiparameter cytometric analysis of complex cellular response
Authors ŠIMEČKOVÁ, Šárka (203 Czech Republic, belonging to the institution), Radek FEDR (203 Czech Republic), Ján REMŠÍK (703 Slovakia, belonging to the institution), Z. KAHOUNOVÁ (203 Czech Republic), Eva SLABÁKOVÁ (203 Czech Republic) and Karel SOUČEK (203 Czech Republic, guarantor).
Edition Cytometry Part A, HOBOKEN, John Wiley & Sons, 2018, 1552-4922.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10601 Cell biology
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
WWW URL
Impact factor Impact factor: 3.433
RIV identification code RIV/00216224:14310/18:00102432
Organization unit Faculty of Science
Doi http://dx.doi.org/10.1002/cyto.a.23295
UT WoS 000426061500014
Keywords in English flow cytometry; multiparametric analysis; DNA damage; apoptosis; proliferation; immunophenotyping
Tags International impact, Reviewed
Changed by Changed by: Mgr. Michal Petr, učo 65024. Changed: 23/4/2024 11:07.
Abstract
Complex analysis of cellular responses after experimental treatment is important for screening, mechanistic understanding of treatment effects, and the identification of sensitive and resistant cell phenotypes. Modern multicolor flow cytometry has demonstrated its power for such analyses. Here, we introduce a multiparametric protocol for complex analysis of cytokinetics by the simultaneous detection of seven fluorescence parameters. This analysis includes the detection of two surface markers for immunophenotyping, analysis of proliferation based on the cell cycle and the measurement of incorporated nucleoside analogue 5-ethynyl-2-deoxyuridine (EdU) in newly synthesized DNA, analysis of DNA damage using an anti-phospho-histone H2A.X (Ser139) antibody, and determination of cell death using a fixable viability probe and intracellular detection of caspase-3 activation. To demonstrate the applicability of this protocol for the analysis of heterogeneous and complex cell responses, we used different treatments and model cell lines. We demonstrated that this protocol has the potential to provide complex and simultaneous analysis of cytokinetics and analyze the heterogeneity of the response at the single-cell level. (c) 2017 International Society for Advancement of Cytometry
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