altInjecting Membrane Impermeable Substances into Single Cells

The ability to load membrane impermeable substances into cells is of great interest to cell biologists.  Optical transfection is one of the few technologies that allows single cells to easily be transfected, without affecting neighbouring cells.  The image shows six cells, three of which were transfected with a plasmid encoding for green fluorescent protein (green & blue cells), and three of which that were not.  Cells are co-stained with the blue nuclear dye, DAPI, so only the nuclei of untransfected cells are visible.   Early transfection work employed a Ti:Sapphire femtosecond pulsed laser using a standard Gaussian beam profile.  More recent work has employed the use of a Bessel Beam profile, obviating the need for exact focussing and opening the possibility of a “point-and-shoot” device for the biological community.  Current work is ongoing to apply this technology to an axicon tipped fibre, thereby freeing the operator from bulky objectives.

 

Key References

(If desired, all references and pdfs may be downloaded en masse in an Endnote formatted zip file)

Spatially optimized gene transfection by laser-induced breakdown of optically trapped nanoparticles, Yoshihiko Arita, Maria Leilani Torres-Mapa, Woei Ming Lee, Tomáš Čižmár, Paul Campbell, Frank J. Gunn-Moore & Kishan Dholakia, accepted for publication in Applied Physics Letters.

Optical transfection using an endoscope-like system, Nan Ma, Frank Gunn-Moore and Kishan Dholakia, accepted for publication in the Journal of Biomedical Optics.

Enhancement and optimization of plasmid expression in femtosecond optical transfection, Bavishna B. Praveen, David J. Stevenson, Maciej Antkowiak, Kishan Dholakia and Frank J. Gunn-Moore (article online in advance of print) Journal of Biophotonics (2011) (pdf)

Application of dynamic diffractive optics for enhanced femtosecond laser based cell transfection, Maciej Antkowiak, Maria Leilani Torres-Mapa, Frank Gunn-Moore, Kishan Dholakia. Journal of Biophotonics, 3: 696–705 (2010) (Editor's Choice, October 2010)

Integrated optical transfection system using a microlens fiber combined with microfluidic gene delivery, N. Ma, P. C. Ashok, D. J. Stevenson, F. J. Gunn-Moore, and K. Dholakia, Biomedical Optics Express, 1(2), 694-705 (2010) (article online in advance of print)

Light forces the pace: Optical manipulation for Biophotonics, D.J. Stevenson, F. Gunn-Moore and K. Dholakia, J. Biomed. Opt 15(2), 041503 (2010) (selected for re-publication in SPIE Reviews and the Virtual Journal of Biological Physics Research)

Optical injection of mammalian cells using a microfluidic platform, Robert F. Marchington, Yoshihiko Arita, Xanthi Tsampoula, Frank J. Gunn-Moore, and Kishan Dholakia, Biomedical Optics Express, Vol. 1, Issue 2, pp. 527-536 (2010).

Light beats the spread: “non-diffracting” beams, Michael Mazilu, David J. Stevenson, Frank J. Gunn-Moore and Kishan Dholakia, Laser & Photonics Reviews 4 (4), 529 - 547 (2010).

Quantitative phase study of the dynamic cellular response in femtosecond laser photoporation, Maciej Antkowiak, Maria Leilani Torres-Mapa, Kishan Dholakia, and Frank J. Gunn-Moore. Biomedical Optics Express, Vol. 1, Issue 2, pp. 414-424 (2010).

Photo-transfection of mammalian cells using femtosecond laser pulses: optimisation and applicability to stem cell differentiation Patience Mthunzi, Kishan Dholakia, Frank Gunn-Moore, J. Biomed. Opt 15(2), 041507 (2010) (highlighted in the Virtual Journal of Ultrafast Science)

Transient transfection of mammalian cells using a violet diode laser Maria Leilani Torres-Mapa, Liselotte Veldhuis, Martin Ploschner, Kishan Dholakia, Frank Gunn-Moore, J. Biomed. Opt 15(2), 041506 (2010) (article online in advance of print)

Application of dynamic diffractive optics for enhanced femtosecond based cell transfection M. Antkowiak, L. Torres-Mapa, F. Gunn-Moore, K. Dholakia, article online in advance of print in the Journal of Biophotonics (2010).

Transfection by optical injection David J. Stevenson, Frank J. Gunn-Moore, Paul Campbell, Kishan Dholakia, Chapter 3 in The Handbook of Photonics for Medical Science Ed. by Tuchin V.V. London, CRC Press, Taylor & Francis Group, 2010. pp.87-117. ( This e-mail address is being protected from spambots. You need JavaScript enabled to view it. )

Single cell optical transfection David J. Stevenson, Frank J. Gunn-Moore, Paul Campbell, Kishan Dholakia, Journal of Royal Society Interface 7(47) 863-871. ( This e-mail address is being protected from spambots. You need JavaScript enabled to view it. )

Targeted optical injection of gold nanoparticles into single mammalian cells, C McDougall, D.J. Stevenson, C.T.A. Brown, F. Gunn-Moore, & K. Dholakia. Journal of Biophotonics 2(12) 736–743 (2009) (pdf) (video 1) (video 2) (press highlight: arstechnica.com)

Fibre based cellular transfection X. Tsampoula, K. Taguchi, T. Cižmár, V. Garces-Chavez, N. Ma, S. Mohanty, K. Mohanty, F. Gunn-Moore, and K. Dholakia, Optics Express, 16, 21, 17007-17013 (2008) (pdf)

Enhanced Operation of Femtosecond Lasers and Applications In Cell Transfection
C.T.A. Brown, D.J. Stevenson, X. Tsampoula, C. McDougall, A. Lagatsky, W. Sibbett, F.J. Gunn-Moore and K. Dholakia, Journal of Biophotonics 1(3), 183 - 199 (2008) (pdf) (wmv) (wmv)

Femtosecond cellular transfection using a non-diffracting light beam
X.Tsampoula, V. Garcés-Chávez, M. Comrie, D. Stevenson, M.B. Agate, F.J. Gunn-Moore, C.T.A. Brown and K Dholakia, Appl Phys Lett 91, 053902 (2007). Reprint in Virtual Journal of Biological Physics Research (pdf)

Femtosecond optical transfection of cells: viability and efficiency
D. Stevenson, B. Agate, X. Tsampoula, P. Fischer, C. T. A. Brown, W. Sibbett A. Riches, F. Gunn- Moore, K. Dholakia Opt. Express 14, 7125-7133 (2006) (pdf)

Photoporation and cell transfection using a violet diode laser (Press release)
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, Opt. Express 13, 595-600 (2005) (pdf)

We attempt to maintain, to the best of our ability, the wiki entry on optical transfection.