Phototransfection of mammalian cells using femtosecond laser pulses: Optimization and applicability to stem cell differentiation

Patience Mthunzi; Kishan Dholakia; Frank Gunn-Moore

doi:10.1117/1.3430733

Phototransfection of mammalian cells using femtosecond laser pulses: Optimization and applicability to stem cell differentiation

Patience Mthunzi, Kishan Dholakia, Frank Gunn-Moore

Department of Physics

Research output: Contribution to journal › Article › peer-review

46 Citations (Scopus)

Abstract

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.

Original language	English
Article number	041507
Journal	Journal of Biomedical Optics
Volume	15
Issue number	4
DOIs	https://doi.org/10.1117/1.3430733
Publication status	Published - 2010 Jul

Bibliographical note

Funding Information:
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.

Funding 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

All Science Journal Classification (ASJC) codes

Electronic, Optical and Magnetic Materials
Biomaterials
Atomic and Molecular Physics, and Optics
Biomedical Engineering

Access to Document

10.1117/1.3430733

Cite this

@article{8cd94d1cd7114723a1e48813002b151c,

title = "Phototransfection of mammalian cells using femtosecond laser pulses: Optimization and applicability to stem cell differentiation",

abstract = "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.",

author = "Patience Mthunzi and Kishan Dholakia and Frank Gunn-Moore",

note = "Funding Information: 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. Funding 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",

year = "2010",

month = jul,

doi = "10.1117/1.3430733",

language = "English",

volume = "15",

journal = "Journal of Biomedical Optics",

issn = "1083-3668",

publisher = "SPIE",

number = "4",

}

TY - JOUR

T1 - Phototransfection of mammalian cells using femtosecond laser pulses

T2 - Optimization and applicability to stem cell differentiation

AU - Mthunzi, Patience

AU - Dholakia, Kishan

AU - Gunn-Moore, Frank

N1 - Funding Information: 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. Funding 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

PY - 2010/7

Y1 - 2010/7

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=78651386540&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78651386540&partnerID=8YFLogxK

U2 - 10.1117/1.3430733

DO - 10.1117/1.3430733

M3 - Article

C2 - 20799785

AN - SCOPUS:78651386540

SN - 1083-3668

VL - 15

JO - Journal of Biomedical Optics

JF - Journal of Biomedical Optics

IS - 4

M1 - 041507

ER -

Phototransfection of mammalian cells using femtosecond laser pulses: Optimization and applicability to stem cell differentiation

Abstract

Bibliographical note

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this