Femtosecond-laser induced membrane optoporation for transfection and cell reprogramming

Hans Georg Breunig; Aisada König; Karsten König

doi:10.1117/12.2607258

3 March 2022 Femtosecond-laser induced membrane optoporation for transfection and cell reprogramming

Hans Georg Breunig, Aisada König, Karsten König

Proceedings Volume 11965, Multiphoton Microscopy in the Biomedical Sciences XXII; 119650G (2022) https://doi.org/10.1117/12.2607258
Event: SPIE BiOS, 2022, San Francisco, California, United States

Conference Poster

Abstract

Short irradiation of a cell with focused femtosecond laser pulses can make the membrane of the cell transiently permeable such that macro molecules from the surrounding microenvironment can enter. This laser-induced sub-micron optoporation is well tolerated and makes it possible to efficiently transfect a cell with desired nucleic acids (DNA/RNA) and, thereby, even to reprogram it. However, for high cell viability and high transfection efficiency individual cell targeting is required, which limits the number of addressable cells per time. We have investigated the laser-assisted cell transfection of adherent mammal and human cells by manually and, to increase the throughput, automatically targeting the laser focus onto the cells. A different strategy is to irradiate continuously flowing cells without individual targeting. This allows to increases the number of addressable cells per time, but at the cost of the efficiency, since not all cells are optimally hit. A precise control of the cell flow as well as the focusing conditions is crucial to maximize the laser-cell interaction in that case. This is possible with a microfluidic setup with flow control where the cells pass a light-sheet like focal region which consists of a scanned Bessel beam of femtosecond-laser pulses. We summarize and review our experimental approaches for laser transfection and non-viral optical cell reprogramming to generate induced pluripotent stem cells through the use of ultrashort near infrared laser pulses.

Citation Download Citation

Hans Georg Breunig, Aisada König, and Karsten König "Femtosecond-laser induced membrane optoporation for transfection and cell reprogramming", Proc. SPIE 11965, Multiphoton Microscopy in the Biomedical Sciences XXII, 119650G (3 March 2022); https://doi.org/10.1117/12.2607258

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