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结合多光子显微镜进行单细胞研究的光学操控

Optical manipulation in combination with multiphoton microscopy for single-cell studies.

作者信息

Goksör Mattias, Enger Jonas, Hanstorp Dag

机构信息

Department of Experimental Physics, Chalmers University of Technology and Göteborg University, SE-412 96 Göteborg, Sweden.

出版信息

Appl Opt. 2004 Sep 1;43(25):4831-7. doi: 10.1364/ao.43.004831.

Abstract

We demonstrate how optical tweezers can be incorporated into a multiphoton microscope to achieve three-dimensional imaging of trapped cells. The optical tweezers, formed by a cw 1064 nm Nd:YVO4 laser, were used to trap live yeast cells in suspension while the 4',6-diamidino-2-phenylindole-stained nucleus was imaged in three dimensions by use of a pulsed femtosecond laser. The trapped cell was moved in the axial direction by changing the position of an external lens, which was used to control the divergence of the trapping laser beam. This gives us a simple method to use optical tweezers in the laser scanning of confocal and multiphoton microscopes. It is further shown that the same femtosecond laser as used for the multiphoton imaging could also be used as laser scissors, allowing us to drill holes in the membrane of trapped spermatozoa.

摘要

我们展示了如何将光镊整合到多光子显微镜中,以实现对捕获细胞的三维成像。由连续波1064 nm Nd:YVO4激光形成的光镊,用于捕获悬浮液中的活酵母细胞,同时利用脉冲飞秒激光对4',6-二脒基-2-苯基吲哚染色的细胞核进行三维成像。通过改变外部透镜的位置使捕获的细胞在轴向移动,该外部透镜用于控制捕获激光束的发散。这为我们在共聚焦和多光子显微镜的激光扫描中使用光镊提供了一种简单方法。进一步表明,用于多光子成像的同一飞秒激光也可用作激光剪刀,使我们能够在捕获的精子细胞膜上钻孔。

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