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首页> 外文会议>Conference on Multiphoton Microscopy in the Biomedical Sciences III Jan 26-28, 2003 San Jose, California, USA >The effects of femtosecond pulse dispersion pre-compensation on average power damage thresholds for live cell imaging: implications for relative roles of linear and nonlinear absorption in live cell imaging
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The effects of femtosecond pulse dispersion pre-compensation on average power damage thresholds for live cell imaging: implications for relative roles of linear and nonlinear absorption in live cell imaging

机译：飞秒脉冲色散预补偿对活细胞成像平均功率损伤阈值的影响：对活细胞成像中线性和非线性吸收的相对作用的暗示

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摘要

Multiphoton microscopy is becoming an increasingly popular modality of laser scanning microscopy for imaging living specimens. In order to improve the signal to noise ratio under the challenging imaging conditions typical of biomedical research, researchers may be forced to resort to measures which substantially increase the amount of energy to which specimens are exposed. Several mechanisms of damage to living cells compete to limit the power window for minimally invasive imaging in multiphoton microscopy; these mechanisms include heating due to linear absorption, phototoxicity related to multiphoton absorption, and optical breakdown due to multiphoton ionization. The relative contribution of each of these factors may change significantly depending on the specimen and imaging parameters. The present study investigates the dominant factors in limiting cell viability at moderate to high-energy fluence levels that may be necessary under non-ideal imaging conditions. The results of this study suggest that heating associated with optical breakdown is an important factor in limiting cell viability under difficult imaging conditions, and that this heating scales in a manner proportional to the energy fluence associated with a given set of scanning parameters. Also significant is the finding that cell viability does not appear to scale proportionally to pulse width in a manner consistent with 2-photon absorption at ultrashort pulse widths and moderate energy fluence levels. These results suggest that the use of a pre-compensation scheme to offset the positive group velocity dispersion due to laser scanning microscope optics is a practical means to increase the signal to noise ratio while minimally impacting cell damage thresholds at ultrafast (sub 200 femtosecond) pulse widths.
机译：多光子显微镜正成为用于对活体标本成像的激光扫描显微镜的一种越来越流行的方式。为了在具有挑战性的生物医学研究典型成像条件下提高信噪比，研究人员可能不得不采取措施，大幅增加标本所暴露的能量。几种损伤活细胞的机制竞争着限制多光子显微镜中微创成像的功率窗口。这些机制包括由于线性吸收引起的加热，与多光子吸收有关的光毒性以及由于多光子电离引起的光学击穿。这些因素各自的相对贡献可能会根据样本和成像参数而发生显着变化。本研究调查了在非理想成像条件下可能需要的中等至高能量通量水平下限制细胞活力的主要因素。这项研究的结果表明，与光学击穿相关的加热是限制在艰难成像条件下细胞存活力的重要因素，并且这种加热的规模与与给定扫描参数集相关的能量通量成比例。同样重要的发现是，细胞活力似乎并未以与超短脉冲宽度和中等能量通量水平下的2光子吸收一致的方式与脉冲宽度成比例地缩放。这些结果表明，使用预补偿方案来抵消由于激光扫描显微镜光学器件引起的正群速度色散是增加信噪比，同时在超快（小于200飞秒）脉冲时对细胞损伤阈值的影响最小的实用方法。宽度。

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