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

Phototransfection of mammalian cells using femtosecond laser pulses: optimization and applicability to stem cell differentiation

[+] Author Affiliations
Patience Mthunzi

University of St. Andrews, School of Physics and Astronomy, North Haugh, St. Andrews, Fife, KY16 9SS Scotland, United Kingdom and Biophotonics: National Laser Centre, Council for Scientific and Industrial Research (CSIR), P.O. Box 395, Pretoria, 0001 South Africa

Kishan Dholakia

University of St. Andrews, School of Physics and Astronomy, North Haugh, St. Andrews, Fife, KY16 9SS Scotland, United Kingdom

Frank Gunn-Moore

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

J. Biomed. Opt. 15(4), 041507 (July 06, 2010). doi:10.1117/1.3430733
History: Received October 19, 2009; Revised January 19, 2010; Accepted February 03, 2010; Published July 06, 2010; Online July 06, 2010
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Recently, femtosecond laser pulses have been utilized for the targeted introduction of genetic matter into mammalian cells. This rapidly expanding and developing novel optical technique using a tightly focused laser light beam is called phototransfection. Extending previous studies [Stevenson et al., Opt. Express, 7125–7133 (2006)], we show that femtosecond lasers can be used to phototransfect a range of different cell lines, and specifically that this novel technology can also transfect mouse embryonic stem cell colonies with 25% efficiency. Notably, we show the ability of differentiating these cells into the extraembryonic endoderm using phototransfection. Furthermore, we present two new findings aimed at optimizing the phototransfection method and improving applicability: first, the influence of the cell passage number on the transfection efficiency is explored and, second, the ability to enhance the transfection efficiency via whole culture treatments. Our results should encourage wider uptake of this methodology.

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© 2010 Society of Photo-Optical Instrumentation Engineers

Citation

Patience Mthunzi ; Kishan Dholakia and Frank Gunn-Moore
"Phototransfection of mammalian cells using femtosecond laser pulses: optimization and applicability to stem cell differentiation", J. Biomed. Opt. 15(4), 041507 (July 06, 2010). ; http://dx.doi.org/10.1117/1.3430733


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