Three-photon microscopy has been increasingly adopted for probing neural activities beyond the typical two-photon imaging depth. In this review, we outline the unique properties that differentiate three-photon microscopy from two-photon microscopy for in vivo imaging in biological samples, especially in the mouse brain. We present a systematic summary of the optimization of three-photon imaging parameters for neural imaging, based on their effects on calcium imaging quality and perturbation to brain tissues. Furthermore, we review the existing techniques for volumetric imaging and discuss their prospects in mesoscale three-photon imaging in deep tissue.

中文翻译：

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小鼠深部脑部的三光子神经元成像

三光子显微镜已越来越多地用于探测超出典型的两光子成像深度的神经活动。在这篇综述中，我们概述了区分三光子显微镜和两光子显微镜的独特特性，以便在生物样品中进行体内成像，尤其是在小鼠大脑中。我们根据神经成像的三光子成像参数对钙成像质量和对脑组织的扰动的影响，对三光子成像参数的优化进行了系统总结。此外，我们回顾了现有的体积成像技术，并讨论了它们在深层组织中尺度三光子成像中的前景。