MICROMANIPULATION OF CHROMOSOMES IN PTK2 CELLS USING LASER MICROSURGERY (OPTICAL SCALPEL) IN COMBINATION WITH LASER-INDUCED OPTICAL FORCE (OPTICAL TWEEZERS)
MICROMANIPULATION OF CHROMOSOMES IN PTK2 CELLS USING LASER MICROSURGERY (OPTICAL SCALPEL) IN COMBINATION WITH LASER-INDUCED OPTICAL FORCE (OPTICAL TWEEZERS)
复制标题
DOI：
10.1006/excr.1993.1015
复制
发表时间：
1993-01-01
影响因子：
3.7
通讯作者：
BERNS, MW
中科院分区：
文献类型：
--
作者：
LIANG, H;WRIGHT, WH;BERNS, MW
光手术刀和光镊相结合，可进行细胞内显微手术和体内显微操作。当仅对中期染色体进行激光显微手术时，解剖的姐妹染色单体片段会漂移到纺锤体的一侧或完全脱离纺锤体。在后期，碎片分离，两条臂通常移动到各自的子细胞。当染色体臂在后期 A 和 B 期间被切断时，远端染色体片段与染色体的其余部分分离，并跟随近端染色体片段向极移动。在中期激光切割的远端染色体片段在整个后期使用光陷阱保持在一起。后期 A 和 B 期间解剖的染色体片段的光学捕获抑制了染色体片段向其极点的移动。结果，捕获的染色体片段（1）并入相对的子细胞，（2）在胞质分裂过程中在卵裂沟中丢失，或（3）最终并入正确的子细胞。这些结果表明，光陷阱可以有效地在整个有丝分裂过程中保留激光切割的染色体片段。这种新工具对于染色体运动和细胞遗传学的研究应该很有用。
An optical scalpel and optical tweezer have been combined to perform intracellular microsurgery and micromanipulationin 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.