Micromanipulation of mitotic chromosomes in PTK2 cells using laser-induced optical forces ("optical tweezers").
To study the potential use of optical forces to manipulate chromosome movement, we have used a Nd:YAG laser at a wavelength of 1.06 microns focused into a phase contrast microscope. Metaphase and anaphase chromosomes were exposed while being monitored by video microscopy. The results indicated that when optical forces were applied to late-moving metaphase chromosomes on the side closest to the nearest spindle pole, the trapped chromosomes initiated movement to the metaphase plate.  The chromosome velocities were two to eight times the normal rate depending on the chromosome size, geometry, and trapping site. At the initiation of anaphase, a pair of chromatids could be held by the optical trap and kept motionless throughout anaphase while the other pairs of chromatids separated and moved to opposite spindle poles. As a result, the trapped chromosome either was incorporated into one of the daughter cells or was lost in the cleavage furrow, or the two chromatids eventually separated and moved to their respective daughter cells. If the trap was removed at the beginning of anaphase B, the chromosome moved back to the poles. Our experiments demonstrate that the laser-induced optical force trap is a potential new technique to study noninvasively the mitotic spindle of living cells.
Liang
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Beckman Laser Institute and Medical Clinic, University of California, Irvine, California 92715.
Wright
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He
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Berns
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D000704:Analysis of Variance; D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D002875:Chromosomes / Q000378:metabolism; D003584:Cytological Techniques; D007834:Lasers; D007614:Macropodidae; D008297:Male; D008938:Mitosis; D010784:Photomicrography; D051381:Rats