Photoporation and cell transfection using a violet diode laser

L. Paterson, B. Agate, M. Comrie, R. Ferguson, T. K. Lake, J. E. Morris, A. E. Carruthers, C. T.A. Brown, W. Sibbett, P. E. Bryant, F. Gunn-Moore, A. C. Riches, K. Dholakia

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Abstract

The introduction and subsequent expression of foreign DNA inside living mammalian cells (transfection) is achieved by photoporation with a violet diode laser. We direct a compact 405 nm laser diode source into an inverted optical microscope configuration and expose cells to 0.3 mW for 40 ms. The localized optical power density of ∼1200 MW/m2 is six orders of magnitude lower than that used in femtosecond photoporation (∼104 TW/m 2). The beam perforates the cell plasma membrane to allow uptake of plasmid DNA containing an antibiotic resistant gene as well as the green fluorescent protein (GFP) gene. Successfully transfected cells then expand into clonal groups which are used to create stable cell lines. The use of the violet diode laser offers a new and simple poration technique compatible with standard microscopes and is the simplest method of laser-assisted cell poration reported to date.

Original languageEnglish
Pages (from-to)595-600
Number of pages6
JournalOptics Express
Volume13
Issue number2
DOIs
Publication statusPublished - 2005 Jan

All Science Journal Classification (ASJC) codes

  • Atomic and Molecular Physics, and Optics

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    Paterson, L., Agate, B., Comrie, M., Ferguson, R., Lake, T. K., Morris, J. E., Carruthers, A. E., Brown, C. T. A., Sibbett, W., Bryant, P. E., Gunn-Moore, F., Riches, A. C., & Dholakia, K. (2005). Photoporation and cell transfection using a violet diode laser. Optics Express, 13(2), 595-600. https://doi.org/10.1364/OPEX.13.000595