Influence of cell fixation on chromatin topography

KOZUBEK, Stanislav, Emilie LUKÁŠOVÁ, Jana AMRICHOVÁ, Michal KOZUBEK, Andrea LIŠKOVÁ and Jana ŠLOTOVÁ. Influence of cell fixation on chromatin topography. Analytical Biochemistry. San Diego: Academic Press, 2000, vol. 282, No 1, p. 29-38. ISSN 0003-2697.

Basic information

Original name

Influence of cell fixation on chromatin topography

Authors

KOZUBEK, Stanislav (203 Czech Republic), Emilie LUKÁŠOVÁ (203 Czech Republic), Jana AMRICHOVÁ (203 Czech Republic), Michal KOZUBEK (203 Czech Republic), Andrea LIŠKOVÁ (203 Czech Republic) and Jana ŠLOTOVÁ

Edition

Analytical Biochemistry, San Diego, Academic Press, 2000, 0003-2697

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Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10610 Biophysics

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

abstract

Impact factor

Impact factor: 1.976

RIV identification code

RIV/00216224:14330/00:00002778

Organization unit

Faculty of Informatics

UT WoS

000087777300005

Keywords in English

chromatin structure; cell fixation; confocal laser scanning microscopy; image analysis; interphase nucleus

Tags

cell fixation, chromatin structure, confocal laser scanning microscopy, Image analysis, interphase nucleus

Tags

International impact, Reviewed

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Abstract

V originále

Using in situ hybridization techniques, a fixation step must precede denaturation to prevent disintegration of the chromosomes. The analysis of nuclei fixed by paraformaldehyde, preserving the native structure (three-dimensional or 3D fixation and analysis) has become possible with the development of confocal microscopy; however, the analysis of those fixed by methanol and acetic acid, dehydrating the nuclei (two-dimensional or 2D fixation and analysis), remains a very valuable tool for practical use in diagnostics and also in many cases for research. We compared both types of fixation and analyses using different cell lines and different DNA probes. Fixation of cells by methanol and acetic acid leads to the enlargement of contact of nuclei with the slide surface, resulting in a substantial increase of nuclear diameter, flattening of the nucleus, and consequently to a distortion of gene topology.

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Links

GA202/97/0874, research and development project	Name: Struktura interfázního jádra a její změny po ozáření Investor: Czech Science Foundation, The structure of interphase nucleus and its changes after irradiation
MSM 143300002, plan (intention)	Name: Využití počítačové analýzy obrazu v optické mikroskopii Investor: Ministry of Education, Youth and Sports of the CR, Application of computer image analysis in optical microscopy
NM15, research and development project	Name: Užití interfázní FISH techniky pro vyhledávání lidí se zvýšeným rizikem vzniku zhoubných hemoblastóz Investor: Ministry of Health of the CR, The use interphase FISH for the determination of individuals with elevated risk of the induction of hemoblastoses
VS97031, research and development project	Name: Využití analýzy obrazu při studiu mechanismů vzniku, v diagnostice a pro prevenci závažných onemocnění člověka Investor: Ministry of Education, Youth and Sports of the CR, Image analysis in the study of mechanisms of induction, in diagnosis and for prevention of deleterious human diseases