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Proceedings Paper

Employment of laser-induced fusion of living cells for the study of spatial structure of chromatin
Author(s): Jan Jezek; Stanislav Palsa; Emilie Lukasova; Stanislav Kozubek; Petr Jakl; Mojmir Sery; Alexandr Jonas; Miroslav Liska; Pavel Zemanek
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Paper Abstract

We study the transfer of the cell nucleus and individual chromosomes from one living cell to the other one during their fusion. To achieve this, the nuclei of the two fused cells are stained with different fluorescent dyes which serve as identification markers. The fusion itself is done in an inverted optical microscope by combined system that uses optical tweezers to bring two living cells into contact and optical scalpel to punctuate their membranes at the contact point. This process initiates a fusion of both cells into one hybrid cell containing two nuclei. If the fusion product is viable, these nuclei tend to mix together. The dynamics of the fusion process is then visualized by exciting the fluorescently labeled fusion product with a suitable light source. The time evolution of the mutual position of the fused cell nuclei and their final orientation is traced from a video record of the experiment. The spatial distribution of the nuclear material in the resulting hybrid nucleus is studied by analysis of positions of FISH (fluorescent hybridization in situ) signals of specific genetic loci in automated fluorescence microscope (high resolution cytometer). The obtained results are compared to the signals distribution of FISH in the original cells.

Paper Details

Date Published: 21 November 2003
PDF: 5 pages
Proc. SPIE 5259, 13th Polish-Czech-Slovak Conference on Wave and Quantum Aspects of Contemporary Optics, (21 November 2003); doi: 10.1117/12.545162
Show Author Affiliations
Jan Jezek, Institute of Scientific Instruments (Czech Republic)
Stanislav Palsa, Institute of Biophysics (Czech Republic)
Emilie Lukasova, Institute of Biophysics (Czech Republic)
Stanislav Kozubek, Institute of Biophysics (Czech Republic)
Petr Jakl, Institute of Scientific Instruments (Czech Republic)
Mojmir Sery, Institute of Scientific Instruments (Czech Republic)
Brno Univ. of Technology (Czech Republic)
Alexandr Jonas, Institute of Scientific Instruments (Czech Republic)
Miroslav Liska, Brno Univ. of Technology (Czech Republic)
Pavel Zemanek, Institute of Scientific Instruments (Czech Republic)


Published in SPIE Proceedings Vol. 5259:
13th Polish-Czech-Slovak Conference on Wave and Quantum Aspects of Contemporary Optics

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