Employment of laser-induced fusion of living cells for the study of spatial structure of chromatin

Jan Jezek; Stanislav Palsa; Emilie Lukasova; Stanislav Kozubek; Petr Jakl; Mojmir Sery; Alexandr Jonas; Miroslav Liska; Pavel Zemanek

doi:10.1117/12.545162

21 November 2003 Employment of laser-induced fusion of living cells for the study of spatial structure of chromatin

Jan Jezek, Stanislav Palsa, Emilie Lukasova, Stanislav Kozubek, Petr Jakl, Mojmir Sery, Alexandr Jonas, Miroslav Liska, Pavel Zemanek

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Proceedings Volume 5259, 13th Polish-Czech-Slovak Conference on Wave and Quantum Aspects of Contemporary Optics; (2003) https://doi.org/10.1117/12.545162
Event: 13th Polish-Czech-Slovak Conference on Wave and Quantum Aspects of contemporary Optics, 2002, Krzyzowa, Poland

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.

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Jan Jezek, Stanislav Palsa, Emilie Lukasova, Stanislav Kozubek, Petr Jakl, Mojmir Sery, Alexandr Jonas, Miroslav Liska, and Pavel Zemanek "Employment of laser-induced fusion of living cells for the study of spatial structure of chromatin", Proc. SPIE 5259, 13th Polish-Czech-Slovak Conference on Wave and Quantum Aspects of Contemporary Optics, (21 November 2003); https://doi.org/10.1117/12.545162

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KEYWORDS

Optical tweezers

Laser welding

Microscopes

Telescopes

Ultraviolet radiation

Lenses

Objectives

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