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Research Papers

Spatially sculpted laser scissors for study of DNA damage and repair

[+] Author Affiliations
Jared Stephens, Samarendra K. Mohanty, Suzanne Genc

University of California, Irvine, Beckman Laser Institute and Medical Clinic, 1002 Health Sciences Road East, Irvine, California 92617

Xiangduo Kong, Kyoko Yokomori

University of California, Irvine, School of Medicine, Department of Biological Chemistry, 240D Med Sci I, Irvine, California 92697

Michael W. Berns

University of California, Irvine, Beckman Laser Institute and Medical Clinic, 1002 Health Sciences Road East, Irvine, California 92617 andUniversity of California, Irvine, Department of Biomedical Engineering, 3120 Natural Sciences II, Irvine, California 92697

J. Biomed. Opt. 14(5), 054004 (September 22, 2009). doi:10.1117/1.3213601
History: Received March 25, 2009; Revised June 09, 2009; Accepted July 06, 2009; Published September 22, 2009
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We present a simple and efficient method for controlled linear induction of DNA damage in live cells. By passing a pulsed laser beam through a cylindrical lens prior to expansion, an elongated elliptical beam profile is created with the ability to expose controlled linear patterns while keeping the beam and the sample stationary. The length and orientation of the beam at the sample plane were reliably controlled by an adjustable aperture and rotation of the cylindrical lens, respectively. Localized immunostaining by the DNA double strand break (DSB) markers phosphorylated H2AX (γH2AX) and Nbs1 in the nuclei of HeLa cells exposed to the “line scissors” was shown via confocal imaging. The line scissors method proved more efficient than the scanning mirror and scanning stage methods at induction of DNA DSB damage with the added benefit of having a greater potential for high throughput applications.

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© 2009 Society of Photo-Optical Instrumentation Engineers

Citation

Jared Stephens ; Samarendra K. Mohanty ; Suzanne Genc ; Xiangduo Kong ; Kyoko Yokomori, et al.
"Spatially sculpted laser scissors for study of DNA damage and repair", J. Biomed. Opt. 14(5), 054004 (September 22, 2009). ; http://dx.doi.org/10.1117/1.3213601


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