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Laser assisted transfection and manipulation of cells in a microfluidic setup

激光辅助微流体装置中的细胞转染和操作

基本信息

批准号：
285718390
负责人：
Professor Dr. Karsten König
金额：
--
依托单位：
Lehrstuhl für Biophotonik und Lasertechnologie
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2017
资助国家：
德国
起止时间：
2016-12-31 至 2021-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/285718390?language=en
关键词：
Laser assisted transfection manipulation cells

项目摘要

The goal of the project is to realize a novel automated setup for laser-assisted cell membrane poration and transfection of flowing cells in a microfluidic setup. The microfluidic setup spatially confines the cells and guides them through a focused laser beam with variable focal length. Through the interaction with the radiation, the membranes of the cells become shortly and transiently permeable ("nanohole") while the cells pass the laser beam. In this way macro molecules from the surrounding liquid can reach the cytoplasm. The novel setup combines the advantages of laser-assisted cell transfection with microfluidic and avoids the usually tedious and time-consuming exact focusing of the laser light on single cells. In this way the novel setup will allow to perform laser-assisted cell transfection with high cell throughput and high cell viability. The microfluidic setup allows a precise control of the flow velocity, with fast stop and start of the flow at microsecond switching times, the diagnosis of a successful transfection as well as the sorting out of non-transfected cells for further laser illumination or the complete removal from the cell population. A successful transfection process is diagnosed in a later second run based on fluorescence detection for example from green fluorescent protein (GFP) expression. In case of a successful project flow the setup will be integrated into a lab-on-a-chip system. In that case the femtosecond laser radiation will be guided by optical fibers to the microfluidic system.

该项目的目标是实现一种新型的自动化装置，用于激光辅助细胞膜穿孔和微流体装置中流动细胞的转染。微流体装置在空间上限制细胞，并引导它们通过具有可变焦距的聚焦激光束。通过与辐射的相互作用，当细胞通过激光束时，细胞的膜变得短暂且瞬时地可渗透（“纳米孔”）。通过这种方式，周围液体中的大分子可以到达细胞质。这种新的装置结合了激光辅助细胞转染与微流体的优点，避免了通常繁琐耗时的激光精确聚焦在单个细胞上。以这种方式，新的设置将允许进行具有高细胞通量和高细胞活力的激光辅助细胞转染。微流体设置允许精确控制流速，在微秒切换时间快速停止和开始流动，诊断成功转染以及分选出未转染的细胞用于进一步激光照射或从细胞群中完全去除。在随后的第二次运行中，基于荧光检测，例如来自绿色荧光蛋白（GFP）表达的荧光检测，诊断成功的转染过程。在项目流程成功的情况下，该设置将集成到芯片实验室系统中。在这种情况下，飞秒激光辐射将由光纤引导到微流体系统。