Hybrid detectors improved time-lapse confocal microscopy of PML
and 53BP1 nuclear body co-localization in DNA lesions

J 2013

Hybrid detectors improved time-lapse confocal microscopy of PML and 53BP1 nuclear body co-localization in DNA lesions

FOLTÁNKOVÁ, Veronika, Pavel MATULA, Dmitry SOROKIN, Stanislav KOZUBEK, Eva BÁRTOVÁ et. al.

Basic information

Original name

Hybrid detectors improved time-lapse confocal microscopy of PML and 53BP1 nuclear body co-localization in DNA lesions

Authors

FOLTÁNKOVÁ, Veronika (203 Czech Republic), Pavel MATULA (203 Czech Republic, guarantor, belonging to the institution), Dmitry SOROKIN (643 Russian Federation, belonging to the institution), Stanislav KOZUBEK (203 Czech Republic) and Eva BÁRTOVÁ (203 Czech Republic)

Edition

Microscopy and Microanalysis, Saarbrücken, Cambridge University Press, 2013, 1431-9276

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

Genetics and molecular biology

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 2.161

RIV identification code

RIV/00216224:14330/13:00065961

Organization unit

Faculty of Informatics

DOI

http://dx.doi.org/10.1017/S1431927612014353

UT WoS

000316221500013

Keywords in English

DNA damage;chromatin; 53BP1; PML bodies; hybrid detectors; confocal microscopy

Abstract

V originále

We used hybrid detectors (HyDs) to monitor the trajectories and interactions of promyelocytic leukemia (GFP-PML) nuclear bodies (NBs) and mCherry-53BP1-positive DNA lesions. 53BP1 protein accumulates in NBs that occur spontaneously in the genome or in gamma-irradiationinduced foci. When we induced local DNA damage by ultraviolet irradiation, we also observed accumulation of 53BP1 proteins into discrete bodies, instead of the expected dispersed pattern. In comparison with photomultiplier tubes (PMTs), which are used for standard analysis by confocal laser scanning microscopy, HyDs significantly eliminated photobleaching of GFP and mCherry fluorochromes during image acquisition. The low laser intensities used for HyD-based confocal analysis enabled us to observe NBs for the longer time periods, necessary for studies of the trajectories and interactions of PML and 53BP1 NBs. To further characterize protein interactions, we used resonance scanning and a novel bioinformatics approach to register and analyze the movements of individual PML and 53BP1 NBs. The combination of improved HyD-based confocal microscopy with a tailored bioinformatics approach enabled us to reveal damage-specific properties of PML and 53BP1 NBs.

Links

GBP302/12/G157, research and development project

Name: Dynamika a organizace chromosomů během buněčného cyklu a při diferenciaci v normě a patologii

Investor: Czech Science Foundation

Citovat

FOLTÁNKOVÁ, Veronika, Pavel MATULA, Dmitry SOROKIN, Stanislav KOZUBEK and Eva BÁRTOVÁ. Hybrid detectors improved time-lapse confocal microscopy of PML and 53BP1 nuclear body co-localization in DNA lesions. Microscopy and Microanalysis. Saarbrücken: Cambridge University Press, 2013, Volume 19, Issue 2, p. 360-369. ISSN 1431-9276. Available from: https://dx.doi.org/10.1017/S1431927612014353.

@article{1076529,
   author = {Foltánková, Veronika and Matula, Pavel and Sorokin, Dmitry and Kozubek, Stanislav and Bártová, Eva},
   article_location = {Saarbrücken},
   article_number = {Issue 2},
   doi = {http://dx.doi.org/10.1017/S1431927612014353},
   keywords = {DNA damage;chromatin; 53BP1; PML bodies; hybrid detectors; confocal microscopy},
   language = {eng},
   issn = {1431-9276},
   journal = {Microscopy and Microanalysis},
   note = {Accepted.},
   title = {Hybrid detectors improved time-lapse confocal microscopy of PML and 53BP1 nuclear body co-localization in DNA lesions},
   url = {http://journals.cambridge.org/action/displayAbstract?fromPage=online&aid=8864223},
   volume = {Volume 19},
   year = {2013}
}

TY  - JOUR
ID  - 1076529
AU  - Foltánková, Veronika - Matula, Pavel - Sorokin, Dmitry - Kozubek, Stanislav - Bártová, Eva
PY  - 2013
TI  - Hybrid detectors improved time-lapse confocal microscopy of PML and 53BP1 nuclear body co-localization in DNA lesions
JF  - Microscopy and Microanalysis
VL  - Volume 19
IS  - Issue 2
SP  - 360-369
EP  - 360-369
PB  - Cambridge University Press
SN  - 14319276
N1  - Accepted.
KW  - DNA damage;chromatin
KW  - 53BP1
KW  - PML bodies
KW  - hybrid detectors
KW  - confocal microscopy
UR  - http://journals.cambridge.org/action/displayAbstract?fromPage=online&aid=8864223
L2  - http://journals.cambridge.org/action/displayAbstract?fromPage=online&aid=8864223
N2  - We used hybrid detectors (HyDs) to monitor the trajectories and interactions of promyelocytic leukemia (GFP-PML) nuclear bodies (NBs) and mCherry-53BP1-positive DNA lesions. 53BP1 protein accumulates in NBs that occur spontaneously in the genome or in gamma-irradiationinduced foci. When we induced local DNA damage by ultraviolet irradiation, we also observed accumulation of 53BP1 proteins into discrete bodies, instead of the expected dispersed pattern. In comparison with photomultiplier tubes (PMTs), which are used for standard analysis by confocal laser scanning microscopy, HyDs significantly eliminated photobleaching of GFP and mCherry fluorochromes during image acquisition. The low laser intensities used for HyD-based confocal analysis enabled us to observe NBs for the longer time periods, necessary for studies of the trajectories and interactions of PML and 53BP1 NBs. To further characterize protein interactions, we used resonance scanning and a novel bioinformatics approach to register and analyze the movements of individual PML and 53BP1 NBs. The combination of improved HyD-based confocal microscopy with a tailored bioinformatics approach enabled us to reveal damage-specific properties of PML and 53BP1 NBs.
ER  -

FOLTÁNKOVÁ, Veronika, Pavel MATULA, Dmitry SOROKIN, Stanislav KOZUBEK and Eva BÁRTOVÁ. Hybrid detectors improved time-lapse confocal microscopy of PML and 53BP1 nuclear body co-localization in DNA lesions. \textit{Microscopy and Microanalysis}. Saarbrücken: Cambridge University Press, 2013, Volume 19, Issue 2, p.~360-369. ISSN~1431-9276. Available from: https://dx.doi.org/10.1017/S1431927612014353.

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