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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 4, Iss. 9 — Sep. 1, 2013
  • pp: 1533–1547

Single-cell optoporation and transfection using femtosecond laser and optical tweezers

Muhammad Waleed, Sun-Uk Hwang, Jung-Dae Kim, Irfan Shabbir, Sang-Mo Shin, and Yong-Gu Lee  »View Author Affiliations

Biomedical Optics Express, Vol. 4, Issue 9, pp. 1533-1547 (2013)

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In this paper, we demonstrate a new single-cell optoporation and transfection technique using a femtosecond Gaussian laser beam and optical tweezers. Tightly focused near-infrared (NIR) femtosecond laser pulse was employed to transiently perforate the cellular membrane at a single point in MCF-7 cancer cells. A distinct technique was developed by trapping the microparticle using optical tweezers to focus the femtosecond laser precisely on the cell membrane to puncture it. Subsequently, an external gene was introduced in the cell by trapping and inserting the same plasmid-coated microparticle into the optoporated cell using optical tweezers. Various experimental parameters such as femtosecond laser exposure power, exposure time, puncture hole size, exact focusing of the femtosecond laser on the cell membrane, and cell healing time were closely analyzed to create the optimal conditions for cell viability. Following the insertion of plasmid-coated microparticles in the cell, the targeted cells exhibited green fluorescent protein (GFP) under the fluorescent microscope, hence confirming successful transfection into the cell. This new optoporation and transfection technique maximizes the level of selectivity and control over the targeted cell, and this may be a breakthrough method through which to induce controllable genetic changes in the cell.

© 2013 OSA

OCIS Codes
(000.1430) General : Biology and medicine
(020.4180) Atomic and molecular physics : Multiphoton processes
(020.7010) Atomic and molecular physics : Laser trapping
(140.7090) Lasers and laser optics : Ultrafast lasers
(320.2250) Ultrafast optics : Femtosecond phenomena
(140.3538) Lasers and laser optics : Lasers, pulsed

ToC Category:
Cell Studies

Original Manuscript: May 2, 2013
Revised Manuscript: June 27, 2013
Manuscript Accepted: June 30, 2013
Published: August 7, 2013

Muhammad Waleed, Sun-Uk Hwang, Jung-Dae Kim, Irfan Shabbir, Sang-Mo Shin, and Yong-Gu Lee, "Single-cell optoporation and transfection using femtosecond laser and optical tweezers," Biomed. Opt. Express 4, 1533-1547 (2013)

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