Recently, femtosecond laser pulses have been utilized for the targeted introduction of genetic matter into mammalian cells. This rapidly expanding and developing novel optical technique using a tightly focused laser light beam is called phototransfection. Extending previous studies [Stevenson et al., Opt. Express 14, 7125-7133 (2006)], we show that femtosecond lasers can be used to phototransfect a range of different cell lines, and specifically that this novel technology can also transfect mouse embryonic stem cell colonies with ∼25% efficiency. Notably, we show the ability of differentiating these cells into the extraembryonic endoderm using phototransfection. Furthermore, we present two new findings aimed at optimizing the phototransfection method and improving applicability: first, the influence of the cell passage number on the transfection efficiency is explored and, second, the ability to enhance the transfection efficiency via whole culture treatments. Our results should encourage Wider uptake of this methodology.
Bibliographical noteFunding Information:
University of St. Andrews School of Physics and Astronomy North Haugh, St. Andrews Fife, KY16 9SS Scotland United Kingdom and Biophotonics: National Laser Centre Council for Scientific and Industrial Research (CSIR) P.O. Box 395 Pretoria, 0001 South Africa
The authors thank the Scottish University Physics Alliance, the Council for Scientific and Industrial Research South Africa, and the UK Engineering and Physical Sciences Research Council for financial support. The authors also thank Dr. Judith Sleeman (University of St. Andrews, School of Biology, Scotland) for providing the E14g2a cells, Dr. Josh Brickman and Maurice Canham (University of Edinburgh, Scottish Centre for Regenerative Medicines, Scotland) for providing the Gata-6 DNA plasmid. Professor Kishan Dholakia is a Royal Society-Wolfson Merit Award Holder.
All Science Journal Classification (ASJC) codes
- Electronic, Optical and Magnetic Materials
- Atomic and Molecular Physics, and Optics
- Biomedical Engineering