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首页> 外文会议>Multiphoton microscopy in the biomedical sciences XI >Photo-induced cell damage analysis for multi-focus CARS microscopy
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Photo-induced cell damage analysis for multi-focus CARS microscopy

机译：光诱导的细胞损伤分析，用于多焦点CARS显微镜

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摘要

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.
机译：我们研究了多聚焦CARS（相干抗斯托克斯拉曼散射）显微镜的光诱导细胞损伤。通常，使用近红外脉冲光源，主要是由多光子引起的现象引起的光致损伤，并且对于非侵入性成像而言，激发光的峰值功率受到限制。我们获得了单焦点或多焦点（7个焦点）曝光期间的细胞活力图像，其波长和脉冲持续时间为709 nm和5 ps。对于单焦点和多焦点激励，将一个焦点的激光功率分别设置为27.8 mW和14.5 mW，因为这些激励光束会感应出与三阶非线性现象相当的信号。使用主要染色死细胞DNA的DAPI荧光团观察到细胞活力。结果，我们发现以27.8 mW /点的单焦点激发在6分钟内引起细胞损伤。相比之下，直到20分钟，对于以14.5 mW /点和7个焦点的多焦点激发，都没有检测到光致损伤。结果表明，光致损伤是单焦点激发的一个严重问题，而多焦点激发方法更适合于CARS成像。

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