Mr. Norman M. Baker Profile

Norman M. Baker

Graduate Student at Univ of California San Diego

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Proceedings Article | 11 September 2006 Paper

Mitotic spindle studied using picosecond laser scissors

N. Baker, E. Botvinick, Linda Shi, M. Berns, George Wu

Proceedings Volume 6326, 63262C (2006) https://doi.org/10.1117/12.680965

KEYWORDS: Spindles, Laser ablation, Microscopes, Picosecond phenomena, Microscopy, Mirrors, Luminescence, Cell death, Ultraviolet radiation, Nd:YAG lasers

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In previous studies we have shown that the second harmonic 532 nm, from a picosecond frequency doubled Nd:YAG laser, can cleanly and selectively disrupt spindle fiber microtubules in live cells (Botvinick et al 2004, Biophys. J. 87:4303-4212). In the present study we have ablated different locations and amounts of the metaphase mitotic spindle, and followed the cells in order to observe the fate of the irradiated spindle and the ability of the cell to continue through mitosis. Cells of the rat kangaroo line (PTK2) were stably transfected by ECFP-tubulin and, using fluorescent microscopy and the automated RoboLase microscope, (Botvinick and Berns, 2005, Micros. Res. Tech. 68:65-74) brightly fluorescent individual cells in metaphase were irradiated with 0.2447 nJ/micropulse corresponding to an irradiance of 1.4496*10^7 J/(ps*cm^2) . Upon irradiation the exposed part of the mitotic spindle immediately lost fluorescence and the following events were observed in the cells over time: (1) immediate contraction of the spindle pole towards the cut, (2) recovery of connection between pole and cut microtubule, (3) completion of mitosis. This system should be very useful in studying internal cellular dynamics of the mitotic spindle.

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