Femtosecond optical transfection of individual mammalian cells

MacIej Antkowiak, Maria L. Torres-Mapa, David J. Stevenson, Kishan Dholakia, Frank J. Gunn-Moore

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Laser-mediated gene transfection into mammalian cells has recently emerged as a powerful alternative to more traditional transfection techniques. In particular, the use of a femtosecond-pulsed laser operating in the near-infrared (NIR) region has been proven to provide single-cell selectivity, localized delivery, low toxicity and consistent performance. This approach can easily be integrated with advanced multimodal live-cell microscopy and micromanipulation techniques. The efficiency of this technique depends on an understanding by the user of both biology and physics. Therefore, in this protocol we discuss the subtleties that apply to both fields, including sample preparation, alignment and calibration of laser optics and their integration into a microscopy platform. The entire protocol takes ∼5 d to complete, from the initial setup of the femtosecond optical transfection system to the final stage of fluorescence imaging to assay for successful expression of the gene of interest.

Original languageEnglish
Pages (from-to)1216-1233
Number of pages18
JournalNature Protocols
Volume8
Issue number6
DOIs
Publication statusPublished - 2013 Jun 1

Fingerprint

Transfection
Microscopic examination
Lasers
Genes
Cells
Laser optics
Microscopy
Pulsed lasers
Optical systems
Micromanipulation
Toxicity
Assays
Optical Devices
Physics
Fluorescence
Optical Imaging
Calibration
Infrared radiation
Imaging techniques
Gene Expression

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Antkowiak, M., Torres-Mapa, M. L., Stevenson, D. J., Dholakia, K., & Gunn-Moore, F. J. (2013). Femtosecond optical transfection of individual mammalian cells. Nature Protocols, 8(6), 1216-1233. https://doi.org/10.1038/nprot.2013.071
Antkowiak, MacIej ; Torres-Mapa, Maria L. ; Stevenson, David J. ; Dholakia, Kishan ; Gunn-Moore, Frank J. / Femtosecond optical transfection of individual mammalian cells. In: Nature Protocols. 2013 ; Vol. 8, No. 6. pp. 1216-1233.
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Antkowiak, M, Torres-Mapa, ML, Stevenson, DJ, Dholakia, K & Gunn-Moore, FJ 2013, 'Femtosecond optical transfection of individual mammalian cells', Nature Protocols, vol. 8, no. 6, pp. 1216-1233. https://doi.org/10.1038/nprot.2013.071

Femtosecond optical transfection of individual mammalian cells. / Antkowiak, MacIej; Torres-Mapa, Maria L.; Stevenson, David J.; Dholakia, Kishan; Gunn-Moore, Frank J.

In: Nature Protocols, Vol. 8, No. 6, 01.06.2013, p. 1216-1233.

Research output: Contribution to journalArticle

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Antkowiak M, Torres-Mapa ML, Stevenson DJ, Dholakia K, Gunn-Moore FJ. Femtosecond optical transfection of individual mammalian cells. Nature Protocols. 2013 Jun 1;8(6):1216-1233. https://doi.org/10.1038/nprot.2013.071