Laser assisted transfection and manipulation of cells in a microfluidic setup

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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.