Raabe I, Vogel S K, Peychl J, Tolić-Nørrelykke I M
Max Planck Institute of Molecular Cell Biology & Genetics, Pfotenhauerstrasse 108, Dresden, Germany.
J Microsc. 2009 Apr;234(1):1-8. doi: 10.1111/j.1365-2818.2009.03142.x.
Living cells are highly organized in space and time, which makes spatially and temporally confined manipulations an indispensable tool in cell biology. Laser-based nanosurgery is an elegant method that allows precise ablation of intracellular structures. Here, we show cutting of fluorescently labelled microtubules and mitotic spindles in fission yeast, performed with a picosecond laser coupled to a confocal microscope. Diverse effects from photo-bleaching to partial and complete breakage are obtained by varying the exposure time, while simultaneously imaging the structures of interest. Using this system we developed an efficient technique to generate enucleated cells without perturbing the distribution of other organelles. This enucleation method can be used to study the cytoskeleton in a nucleus-free environment, as well as the role of the nucleus in cell growth and a variety of cellular functions.
活细胞在空间和时间上高度有序，这使得在空间和时间上受限的操作成为细胞生物学中不可或缺的工具。基于激光的纳米手术是一种优雅的方法，可实现对细胞内结构的精确消融。在此，我们展示了利用与共聚焦显微镜耦合的皮秒激光对裂殖酵母中荧光标记的微管和有丝分裂纺锤体进行切割。通过改变曝光时间，同时对感兴趣的结构进行成像，可获得从光漂白到部分和完全断裂的各种效果。利用该系统，我们开发了一种高效技术，可在不干扰其他细胞器分布的情况下生成去核细胞。这种去核方法可用于在无核环境中研究细胞骨架，以及细胞核在细胞生长和各种细胞功能中的作用。
