Special Section on Selected Topics in Biophotonics: Photodynamic Therapy and Optical Micromanipulation for Biophotonics

Transient transfection of mammalian cells using a violet diode laser

[+] Author Affiliations
Maria Leilani Torres-Mapa

University of St. Andrews, Scottish University Physics Alliance (SUPA), School of Physics and Astronomy, North Haugh, St. Andrews KY16 9SS, Scotland, United Kingdom

Liselotte Angus

University of St. Andrews, School of Biology, Fife, St. Andrews, KY16 9TS, Scotland, United Kingdom

Martin Ploschner, Kishan Dholakia

University of St. Andrews, Scottish University Physics Alliance (SUPA), School of Physics and Astronomy, North Haugh, St. Andrews KY16 9SS, Scotland, United Kingdom

Frank J. Gunn-Moore

University of St. Andrews, School of Biology, Fife, St. Andrews, KY16 9TS, Scotland, United Kingdom

J. Biomed. Opt. 15(4), 041506 (July 07, 2010). doi:10.1117/1.3430730
History: Received September 22, 2009; Revised December 18, 2009; Accepted December 29, 2009; Published July 07, 2010; Online July 07, 2010
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We demonstrate the first use of the violet diode laser for transient mammalian cell transfection. In contrast to previous studies, which showed the generation of stable cell lines over a few weeks, we develop a methodology to transiently transfect cells with an efficiency of up to 40%. Chinese hamster ovary (CHO-K1) and human embryonic kidney (HEK293) cells are exposed to a tightly focused 405-nm laser in the presence of plasmid DNA encoding for a mitochondrial targeted red fluorescent protein. We report transfection efficiencies as a function of laser power and exposure time for our system. We also show, for the first time, that a continuous wave laser source can be successfully applied to selective gene silencing experiments using small interfering RNA. This work is a major step towards an inexpensive and portable phototransfection system.

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© 2010 Society of Photo-Optical Instrumentation Engineers

Citation

Maria Leilani Torres-Mapa ; Liselotte Angus ; Martin Ploschner ; Kishan Dholakia and Frank J. Gunn-Moore
"Transient transfection of mammalian cells using a violet diode laser", J. Biomed. Opt. 15(4), 041506 (July 07, 2010). ; http://dx.doi.org/10.1117/1.3430730


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