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

Patience Mthunzi, Kishan Dholakia, Frank Gunn-Moore

Research output: Contribution to journalArticle

40 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 languageEnglish
Article number041507
JournalJournal of Biomedical Optics
Volume15
Issue number4
DOIs
Publication statusPublished - 2010 Jul 1

Fingerprint

stem cells
Stem cells
Ultrashort pulses
Cells
optimization
pulses
lasers
cells
cultured cells
light beams
mice
laser beams
methodology
Lasers

All Science Journal Classification (ASJC) codes

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

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",
year = "2010",
month = "7",
day = "1",
doi = "10.1117/1.3430733",
language = "English",
volume = "15",
journal = "Journal of Biomedical Optics",
issn = "1083-3668",
publisher = "SPIE",
number = "4",

}

Phototransfection of mammalian cells using femtosecond laser pulses : Optimization and applicability to stem cell differentiation. / Mthunzi, Patience; Dholakia, Kishan; Gunn-Moore, Frank.

In: Journal of Biomedical Optics, Vol. 15, No. 4, 041507, 01.07.2010.

Research output: Contribution to journalArticle

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

PY - 2010/7/1

Y1 - 2010/7/1

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

VL - 15

JO - Journal of Biomedical Optics

JF - Journal of Biomedical Optics

SN - 1083-3668

IS - 4

M1 - 041507

ER -