Photo-induced cell damage analysis for multi-focus CARS microscopy

Takeo Minamikawa; Yoshinori Murakami; Naokazu Matsumura; Hirohiko Niioka; Shuichiro Fukushima; Tsutomu Araki; Mamoru Hashimoto

doi:10.1117/12.874677

11 February 2011 Photo-induced cell damage analysis for multi-focus CARS microscopy

Takeo Minamikawa, Yoshinori Murakami, Naokazu Matsumura, Hirohiko Niioka, Shuichiro Fukushima, Tsutomu Araki, Mamoru Hashimoto

Author Affiliations +

Proceedings Volume 7903, Multiphoton Microscopy in the Biomedical Sciences XI; 79032H (2011) https://doi.org/10.1117/12.874677
Event: SPIE BiOS, 2011, San Francisco, California, United States

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.

Citation Download Citation

Takeo Minamikawa, Yoshinori Murakami, Naokazu Matsumura, Hirohiko Niioka, Shuichiro Fukushima, Tsutomu Araki, and Mamoru Hashimoto "Photo-induced cell damage analysis for multi-focus CARS microscopy", Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032H (11 February 2011); https://doi.org/10.1117/12.874677

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