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使用光镊进行单细胞分离。

Single Cell Isolation Using Optical Tweezers.

作者信息

Keloth Anusha, Anderson Owen, Risbridger Donald, Paterson Lynn

机构信息

Institute of Biological Chemistry, Biophysics and Bioengineering, School of Engineering and Physical Sciences, Heriot Watt University, Edinburgh EH14 4AS, UK.

出版信息

Micromachines (Basel). 2018 Aug 29;9(9):434. doi: 10.3390/mi9090434.

Abstract

Optical tweezers offer a non-contact method for selecting single cells and translocating them from one microenvironment to another. We have characterized the optical tweezing of yeast and can manipulate single cells at 0.41 ± 0.06 mm/s using a 26.8 ± 0.1 mW from a 785 nm diode laser. We have fabricated and tested three cell isolation devices; a micropipette, a PDMS chip and a laser machined fused silica chip and we have isolated yeast, single bacteria and cyanobacteria cells. The most effective isolation was achieved in PDMS chips, where single yeast cells were grown and observed for 18 h without contamination. The duration of budding in was not affected by the laser parameters used, but the time from tweezing until the first budding event began increased with increasing laser energy (laser power × time). Yeast cells tweezed using 25.0 ± 0.1 mW for 1 min were viable after isolation. We have constructed a micro-consortium of yeast cells, and a co-culture of yeast and bacteria, using optical tweezers in combination with the PDMS network of channels and isolation chambers, which may impact on both industrial biotechnology and understanding pathogen dynamics.

摘要

光镊提供了一种非接触式方法,用于挑选单个细胞并将其从一个微环境转移到另一个微环境。我们已经对酵母的光镊操作进行了表征,并且可以使用来自785 nm二极管激光器的26.8±0.1 mW功率,以0.41±0.06 mm/s的速度操纵单个细胞。我们制造并测试了三种细胞分离装置;一个微量移液器、一个聚二甲基硅氧烷(PDMS)芯片和一个激光加工的熔融石英芯片,并且我们已经分离出了酵母、单个细菌和蓝细菌细胞。在PDMS芯片中实现了最有效的分离,在该芯片中,单个酵母细胞生长并观察了18小时而未受污染。酵母出芽的持续时间不受所用激光参数的影响,但从镊子操作到第一次出芽事件开始的时间随着激光能量(激光功率×时间)的增加而增加。使用25.0±0.1 mW的功率镊子操作1分钟后的酵母细胞在分离后仍具有活力。我们使用光镊结合通道和隔离室的PDMS网络构建了酵母细胞的微型聚生体以及酵母和细菌的共培养物,这可能会对工业生物技术和了解病原体动态产生影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2918/6187562/3d1bf6165556/micromachines-09-00434-g001.jpg

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