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J Biomed Opt. 2010 Jul-Aug;15(4):041506. doi: 10.1117/1.3430730.

Transient transfection of mammalian cells using a violet diode laser.

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1
University of St. Andrews, Scottish University Physics Alliance (SUPA), School of Physics and Astronomy, North Haugh, St. Andrews, Scotland, United Kingdom. mlt9@st-andrews.ac.uk

Abstract

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 approximately 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.

PMID:
20799784
DOI:
10.1117/1.3430730
[Indexed for MEDLINE]
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