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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 16, Iss. 5 — Mar. 3, 2008
  • pp: 3021–3031

Quantified femtosecond laser based opto-perforation of living GFSHR-17 and MTH53a cells

J. Baumgart, W. Bintig, A. Ngezahayo, S. Willenbrock, H. Murua Escobar, W. Ertmer, H. Lubatschowski, and A. Heisterkamp  »View Author Affiliations


Optics Express, Vol. 16, Issue 5, pp. 3021-3031 (2008)
http://dx.doi.org/10.1364/OE.16.003021


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Abstract

Opto-perforation is an interesting alternative to conventional techniques for gene transfer into living cells. The cell membrane is perforated by femtosecond (fs) laser pulses, in order to induce an uptake of macromolecules e.g. DNA. In this study, we successfully transfected a canine cell line (MTH53a) with GFP vector or a vector coding for a GFP-HMGB1 fusion protein. The transfected cells were observed 48 hours after treatment and they were not showing any signs of apoptosis or necrosis. Based on simultaneously measured membrane potential changes during the perforation, we were able to calculate and experimentally verify that the relative volume exchanged is 0.4 times the total cell volume. Thus, for first time a quantitative predication of the amount of uptaken molecules and therefore a quantification of the transfection is possible. Additionally, this method offers new high efficient possibilities for critical transfection approaches involving special cell types, e.g. primary and stem cells.

© 2008 Optical Society of America

OCIS Codes
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(320.7090) Ultrafast optics : Ultrafast lasers
(350.4855) Other areas of optics : Optical tweezers or optical manipulation

ToC Category:
Medical Optics and Biotechnology

History
Original Manuscript: January 2, 2008
Revised Manuscript: February 15, 2008
Manuscript Accepted: February 16, 2008
Published: February 20, 2008

Virtual Issues
Vol. 3, Iss. 4 Virtual Journal for Biomedical Optics

Citation
J. Baumgart, W. Bintig, A. Ngezahayo, S. Willenbrock, H. Murua Escobar, W. Ertmer, H. Lubatschowski, and A. Heisterkamp, "Quantified femtosecond laser based opto-perforation of living GFSHR-17 and MTH53 a cells," Opt. Express 16, 3021-3031 (2008)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-16-5-3021


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