Transient transfection of mammalian cells using a violet diode laser

Maria Leilani Torres-Mapa, Liselotte Angus, Martin Ploschner, Kishan Dholakia, Frank J. Gunn-Moore

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

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 similar to 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. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3430730]

Original languageEnglish
Article number041506
Number of pages7
JournalJournal of Biomedical Optics
Volume15
Issue number4
DOIs
Publication statusPublished - 2010

Keywords

  • photoporation
  • phototransfection
  • gene transfection
  • gene knockdown
  • violet diode
  • TARGETED TRANSFECTION
  • FEMTOSECOND LASER
  • VISIBLE-LIGHT
  • GENE-TRANSFER
  • OPTOPORATION
  • GENERATION
  • MEMBRANE

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