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Journal of Biomedical Optics

Phototransfection of mammalian cells using femtosecond laser pulses: optimization and applicability to stem cell differentiation
Author(s): Patience Mthunzi; Kishan Dholakia; Frank James Gunn-Moore
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Paper Abstract

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 14, 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.

Paper Details

Date Published: 1 July 2010
PDF: 7 pages
J. Biomed. Opt. 15(4) 041507 doi: 10.1117/1.3430733
Published in: Journal of Biomedical Optics Volume 15, Issue 4
Show Author Affiliations
Patience Mthunzi, Univ. of St. Andrews (United Kingdom)
Kishan Dholakia, Univ. of St. Andrews (United Kingdom)
Frank James Gunn-Moore, Univ. of St. Andrews (United Kingdom)


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