Towards high-throughput automated targeted femtosecond laser-based transfection of adherent cells

Maciej Antkowiak; Maria Leilani Torres-Mapa; Frank Gunn-Moore; Kishan Dholakia

doi:10.1117/12.874645

11 February 2011 Towards high-throughput automated targeted femtosecond laser-based transfection of adherent cells

Maciej Antkowiak, Maria Leilani Torres-Mapa, Frank Gunn-Moore, Kishan Dholakia

Proceedings Volume 7925, Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications XI; 79250B (2011) https://doi.org/10.1117/12.874645
Event: SPIE LASE, 2011, San Francisco, California, United States

Abstract

Femtosecond laser induced cell membrane poration has proven to be an attractive alternative to the classical methods of drug and gene delivery. It is a selective, sterile, non-contact technique that offers a highly localized operation, low toxicity and consistent performance. However, its broader application still requires the development of robust, high-throughput and user-friendly systems. We present a system capable of unassisted enhanced targeted optoinjection and phototransfection of adherent mammalian cells with a femtosecond laser. We demonstrate the advantages of a dynamic diffractive optical element, namely a spatial light modulator (SLM) for precise three dimensional positioning of the beam. It enables the implementation of a "point-and-shoot" system in which using the software interface a user simply points at the cell and a predefined sequence of precisely positioned doses can be applied. We show that irradiation in three axial positions alleviates the problem of exact beam positioning on the cell membrane and doubles the number of viably optoinjected cells when compared with a single dose. The presented system enables untargeted raster scan irradiation which provides transfection of adherent cells at the throughput of 1 cell per second.

Citation Download Citation

Maciej Antkowiak, Maria Leilani Torres-Mapa, Frank Gunn-Moore, and Kishan Dholakia "Towards high-throughput automated targeted femtosecond laser-based transfection of adherent cells", Proc. SPIE 7925, Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications XI, 79250B (11 February 2011); https://doi.org/10.1117/12.874645

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