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Proceedings Article

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

[+] Author Affiliations
Takeo Minamikawa, Yoshinori Murakami, Naokazu Matsumura, Hirohiko Niioka, Shuichiro Fukushima, Tsutomu Araki, Mamoru Hashimoto

Osaka Univ. (Japan)

Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032H (February 10, 2011); doi:10.1117/12.874677
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From Conference Volume 7903

  • Multiphoton Microscopy in the Biomedical Sciences XI
  • Ammasi Periasamy; Karsten König; Peter T. C. So
  • San Francisco, California, USA | January 22, 2011

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.

© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

Takeo Minamikawa ; Yoshinori Murakami ; Naokazu Matsumura ; Hirohiko Niioka ; Shuichiro Fukushima, et al.
"Photo-induced cell damage analysis for multi-focus CARS microscopy", Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032H (February 10, 2011); doi:10.1117/12.874677; http://dx.doi.org/10.1117/12.874677


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