具有最佳脉冲频率的快速多光子光片显微镜。
Fast multiphoton light-sheet microscopy with optimal pulse frequency.
作者信息
Maioli Vincent, Boniface Antoine, Mahou Pierre, Ortas Júlia Ferrer, Abdeladim Lamiae, Beaurepaire Emmanuel, Supatto Willy
机构信息
Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS, INSERM, Institut Polytechnique de Paris, 91128 Palaiseau, France.
出版信息
Biomed Opt Express. 2020 Oct 1;11(10):6012-6026. doi: 10.1364/BOE.400113.
Abstract
Improving the imaging speed of multiphoton microscopy is an active research field. Among recent strategies, light-sheet illumination holds distinctive advantages for achieving fast imaging . However, photoperturbation in multiphoton light-sheet microscopy remains poorly investigated. We show here that the heart beat rate of zebrafish embryos is a sensitive probe of linear and nonlinear photoperturbations. By analyzing its behavior with respect to laser power, pulse frequency and wavelength, we derive guidelines to find the best balance between signal and photoperturbation. We then demonstrate one order-of-magnitude signal enhancement over previous implementations by optimizing the laser pulse frequency. These results open new opportunities for fast live tissue imaging.
摘要
提高多光子显微镜的成像速度是一个活跃的研究领域。在最近的策略中，光片照明在实现快速成像方面具有独特优势。然而，多光子光片显微镜中的光扰动仍未得到充分研究。我们在此表明，斑马鱼胚胎的心跳速率是线性和非线性光扰动的敏感探针。通过分析其相对于激光功率、脉冲频率和波长的行为，我们得出了在信号和光扰动之间找到最佳平衡的指导原则。然后，我们通过优化激光脉冲频率，证明信号比以前的实现方式提高了一个数量级。这些结果为快速活组织成像开辟了新机会。
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