Multiparameter cytometric analysis of complex cellular response

J 2018

Multiparameter cytometric analysis of complex cellular response

ŠIMEČKOVÁ, Šárka, Radek FEDR, Ján REMŠÍK, Z. KAHOUNOVÁ, Eva SLABÁKOVÁ et. al.

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

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10601 Cell biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

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

Abstract

V originále

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

Citovat

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.

@article{1411854,
   author = {Šimečková, Šárka and Fedr, Radek and Remšík, Ján and Kahounová, Z. and Slabáková, Eva and Souček, Karel},
   article_location = {HOBOKEN},
   article_number = {2},
   doi = {http://dx.doi.org/10.1002/cyto.a.23295},
   keywords = {flow cytometry; multiparametric analysis; DNA damage; apoptosis; proliferation; immunophenotyping},
   language = {eng},
   issn = {1552-4922},
   journal = {Cytometry Part A},
   title = {Multiparameter cytometric analysis of complex cellular response},
   url = {http://dx.doi.org/10.1002/cyto.a.23295},
   volume = {93A},
   year = {2018}
}

TY  - JOUR
ID  - 1411854
AU  - Šimečková, Šárka - Fedr, Radek - Remšík, Ján - Kahounová, Z. - Slabáková, Eva - Souček, Karel
PY  - 2018
TI  - Multiparameter cytometric analysis of complex cellular response
JF  - Cytometry Part A
VL  - 93A
IS  - 2
SP  - 239-248
EP  - 239-248
PB  - John Wiley & Sons
SN  - 15524922
KW  - flow cytometry
KW  - multiparametric analysis
KW  - DNA damage
KW  - apoptosis
KW  - proliferation
KW  - immunophenotyping
UR  - http://dx.doi.org/10.1002/cyto.a.23295
N2  - 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
ER  -

ŠIMEČKOVÁ, Šárka, Radek FEDR, Ján REMŠÍK, Z. KAHOUNOVÁ, Eva SLABÁKOVÁ and Karel SOUČEK. Multiparameter cytometric analysis of complex cellular response. \textit{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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