Single-cell microsurgery: Its problems and possible solutions

Resumo

Generally accepted methods of single-cell microsurgery in terms of their compliance with the task of preserving cell integrity and minimizing possible damage in various microsurgery manipulations on embryos and somatic cells, namely, during microsurgical transfer of nuclei for cloning, production of monozygotic twins, and microinjections into the cell, are analyzed in detail. The problems regarding the cell contacts with microtools, perforation as a factor of cell damage (when a microtool penetrates a cell), the influence of light on the cell, and retention of the cell during microsurgery (and the resulting effect of negative pressure on the cell) are discussed. The question of how long a single cell can persist in a microchamber in vitro is considered, as well as the impact of mineral oil used as a barrier against solvent evaporation. The effect of the physicochemical composition of the glass that is used for microtools is addressed for the first time, as well as the impact of thermal shock and fresh culture medium on a living cell.