Micromanipulation of Chromosomes in PTK2 Cells Using Laser Microsurgery (Optical Scalpel) in Combination with Laser-Induced Optical Force (Optical Tweezers)

Abstract

An optical scalpel and optical tweezer have been combined to perform intracellular microsurgery and micromanipulation in vivo. When only laser microsurgery was performed on metaphase chromosomes, the dissected sister chromatid fragments drifted off to either the side of the spindle or completely off the spindle. At anaphase the fragments separated and the two arms generally moved to their respective daughter cells. When the chromosome arm was cut during anaphase A and B, the distal chromosome fragment separated from the rest of the chromosome and moved toward the pole, following the proximal chromosome fragment. Distal chromosome fragments laser-dissected during metaphase were held together throughout anaphase using the optical trap. Optical trapping of dissected chromosome fragments during anaphase A and B inhibited movement of the chromosome fragment to its pole. As a result, the trapped chromosome fragments were (1) incorporated into the opposite daughter cell, (2) lost in the cleavage furrow during cytokinesis, or (3) eventually incorporated into the correct daughter cell. These results indicate that optical traps are effective in holding laser-dissected chromosome fragments throughout mitosis. This new tool should be useful for studies on chromosome movement and cell genetics.