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Automated adherent cell elimination by a high-speed laser mediated by a light-responsive polymer

机译:由光响应性聚合物介导的高速激光自动消除粘附细胞

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摘要

Conventional cell handling and sorting methods require manual labor, which decreases both cell quality and quantity. To purify adherent cultured cells, cell purification technologies that are high throughput without dissociation and can be utilized in an on-demand manner are expected. Here, we developed a Laser-induced, Light-responsive-polymer-Activated, Cell Killing (LiLACK) system that enables high-speed and on-demand adherent cell sectioning and purification. This system employs a visible laser beam, which does not kill cells directly, but induces local heat production through the trans-cis-trans photo-isomerization of azobenzene moieties. Using this system in each passage for sectioning, human induced pluripotent stem cells (hiPSCs) maintained their pluripotency and self-renewal during long-term culture. Furthermore, combined with deep machine-learning analysis on fluorescent and phase contrast images, a label-free and automatic cell processing system has been developed by eliminating unwanted spontaneously differentiated cells in undifferentiated hiPSC culture conditions.
机译:传统的细胞处理和分选方法需要人工,这降低了细胞的质量和数量。为了纯化贴壁培养的细胞,期望具有高产量而无需解离并且可以按需使用的细胞纯化技术。在这里,我们开发了一种激光诱导的,光响应聚合物激活的细胞杀灭(LiLACK)系统,该系统可实现高速和按需的贴壁细胞切片和纯化。该系统使用可见的激光束,该激光束不会直接杀死细胞,而是通过偶氮苯部分的顺-反-顺光异构化引起局部热量产生。在每个切片中使用此系统进行切片,人类诱导的多能干细胞(hiPSC)在长期培养过程中保持了其多能性和自我更新。此外,结合对荧光和相衬图像的深度机器学习分析,通过消除在未分化的hiPSC培养条件下不需要的自发分化的细胞,开发了一种无标记的自动细胞处理系统。

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