A laser microbeam system has been set up for microsurgery on cell. The relations of laser wavelength, pulse duration and pulse energy to punching effects and self-healing are studied. The experimental results demonstrate that picosecond pulse laser microbeam offers many advantages in cell microsurgery. The mechanism of punching by picosecond microbeam is high field puncture instead of heat effect, and is irrelevant to cell kinds and colors. The diameter and depth of microsurgery can therefore be easily controlled by adjusting the laser pulse energy. The diameter of the minimum aperture is about 0.1 micrometers , much smaller than the theoretical limit ((lambda) /2) for optical microscope due to self- focusing effect. With ultrashort pulse laser microbeam, we can easily cut off any part of a cell. An example is that eight nuclei in the center of unicellular parasite Pneumocystis Carinii can be destroyed one by one by ultrashort pulse laser microbeam without cell wall injury. The holes can also be punched by ultrashort pulse laser microbeam from cell wall to cell nucleus. In a fraction of a second to several seconds after punching, the hole on cell wall or cell membrane can self-heal. Exogenous DNA can be introduced into the cell before its self- healing.
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