Transfection of cells using a violet diode laser for photoporation

Ben Agate, Lynn Paterson, Tanya K. Lake, J. E. Morris, A. E. Carruthers, Christian T. A. Brown, Wilson Sibbett, Kishan Dholakia, Muriel Comrie, R. Ferguson, P. E. Bryant, Andrew C. Riches, Frank J. Gunn-Moore

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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/m2). The tightly focused laser beam (with a localised focal volume of ~10-19 m3 ) 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
Title of host publication2005 Conference on Lasers and Electro-Optics Europe
PublisherOptical Society of America
Pages634
Number of pages1
ISBN (Print)0780389743, 9780780389748
DOIs
Publication statusPublished - 2005
Event2005 Conference on Lasers and Elctro-Optics Europe - Munich, Germany
Duration: 12 Jun 200517 Jun 2005

Conference

Conference2005 Conference on Lasers and Elctro-Optics Europe
CountryGermany
CityMunich
Period12/06/0517/06/05

ASJC Scopus subject areas

  • Control and Systems Engineering
  • Electrical and Electronic Engineering

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  • Cite this

    Agate, B., Paterson, L., Lake, T. K., Morris, J. E., Carruthers, A. E., Brown, C. T. A., Sibbett, W., Dholakia, K., Comrie, M., Ferguson, R., Bryant, P. E., Riches, A. C., & Gunn-Moore, F. J. (2005). Transfection of cells using a violet diode laser for photoporation. In 2005 Conference on Lasers and Electro-Optics Europe (pp. 634). Optical Society of America. https://doi.org/10.1109/CLEOE.2005.1568410