Optical transfection of mammalian cells

David Stevenson; Ben Agate; Lynn Paterson; Tanya Lake; Muriel Comrie; Tom Brown; Andrew Riches; Peter Bryant; Wilson Sibbett; Frank Gunn-Moore; Kishan Dholakia

doi:10.1117/12.662325

14 April 2006 Optical transfection of mammalian cells

David Stevenson, Ben Agate, Lynn Paterson, Tanya Lake, Muriel Comrie, Tom Brown, Andrew Riches, Peter Bryant, Wilson Sibbett, Frank Gunn-Moore, Kishan Dholakia

Author Affiliations +

Proceedings Volume 6191, Biophotonics and New Therapy Frontiers; 61910D (2006) https://doi.org/10.1117/12.662325
Event: SPIE Photonics Europe, 2006, Strasbourg, France

Abstract

The introduction of naked DNA or other membrane impermeable substances into a cell (transfection) is a ubiquitous problem in cell biology. This problem is particularly challenging when it is desired to load membrane impermeable substances into specific cells, as most transfection technologies (such as liposomal transfection) are based on treating a global population of cells. The technique of optical transfection, using a focused laser to open a small transient hole in the membrane of a biological cell (photoporation) to load membrane impermeable DNA into it, allows individual cells to be targeted for transfection, while leaving neighbouring cells unaffected. Unlike other techniques used to perform single cell transfection, such as microinjection, optical transfection can be performed in an entirely closed system, thereby maintaining sterility of the sample during treatment. Here, we are investigating the introduction and subsequent expression of foreign DNA into living mammalian cells by laser-assisted photoporation with a femtosecond-pulsed titanium sapphire laser at 800 nm, in cells that are adherent.

Citation Download Citation

David Stevenson, Ben Agate, Lynn Paterson, Tanya Lake, Muriel Comrie, Tom Brown, Andrew Riches, Peter Bryant, Wilson Sibbett, Frank Gunn-Moore, and Kishan Dholakia "Optical transfection of mammalian cells", Proc. SPIE 6191, Biophotonics and New Therapy Frontiers, 61910D (14 April 2006); https://doi.org/10.1117/12.662325

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