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Proceedings Paper

Photo-induced cell damage analysis for multi-focus CARS microscopy
Author(s): Takeo Minamikawa; Yoshinori Murakami; Naokazu Matsumura; Hirohiko Niioka; Shuichiro Fukushima; Tsutomu Araki; Mamoru Hashimoto
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Paper Abstract

We investigated photo-induced cell damage for multi-focus CARS (coherent anti-Stokes Raman scattering) microscopy. In general, using a near-infrared pulse light source, photo-induced damage is dominantly caused via multi-photon induced phenomena, and the peak power of the excitation light is limited for the non-invasive imaging. We obtained cell viability images during single- or multi-focus (7 foci) exposure of which wavelength and pulse duration were 709 nm and 5 ps. The laser power of one focal spot was respectively set to 27.8 mW and 14.5 mW for single- and multi-focus excitation because those excitation beams induce the comparable signals for third-order nonlinear phenomena. The cell viability was observed using DAPI fluorophore that mainly stains DNA of dead cells. As a result, we found that the single-focus excitation with 27.8 mW/spot caused cell damage within 6 min. In contrast, photo-induced damage was not detected until 20 min for the multi-focus excitation with 14.5 mW/spot and 7 foci. The results suggest that the photo-induced damage is a serious problem on the single-focus excitation, and the multi-focus excitation method is preferable for CARS imaging.

Paper Details

Date Published: 11 February 2011
PDF: 7 pages
Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032H (11 February 2011); doi: 10.1117/12.874677
Show Author Affiliations
Takeo Minamikawa, Osaka Univ. (Japan)
Yoshinori Murakami, Osaka Univ. (Japan)
Naokazu Matsumura, Osaka Univ. (Japan)
Hirohiko Niioka, Osaka Univ. (Japan)
Shuichiro Fukushima, Osaka Univ. (Japan)
Tsutomu Araki, Osaka Univ. (Japan)
Mamoru Hashimoto, Osaka Univ. (Japan)

Published in SPIE Proceedings Vol. 7903:
Multiphoton Microscopy in the Biomedical Sciences XI
Ammasi Periasamy; Karsten König; Peter T. C. So, Editor(s)

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