Two-photon optogenetics has become an indispensable technology in neuroscience, due to its capability in precise and specific manipulation of neural activities. A scanless holographic approach is generally adopted to meet the requirement of stimulating neural ensembles simultaneously. However, the commonly used disk patterns fail in achieving single-neuron resolution, especially in axial dimension, and their inherent speckles decrease stimulation efficiency. Here, we propose a novel speckle-free, beaded-ring pattern for high-axial-resolution optical stimulation of neurons in vivo. Using a dye pool and a fluorescent thin film as samples, we verify that, compared to those with disk patterns, higher axial resolution and better localization ability can be achieved with beaded-ring patterns. Furthermore, we perform two-photon based all-optical physiology with neurons in mouse S1 cortex in vivo, and demonstrate that the axial resolution obtained by beaded-ring patterns can be improved by 24% when stimulating multiple neurons, compared to that of disk patterns.

中文翻译：

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通过具有无斑点珠环图案的双光子光遗传学对体内神经元进行高分辨率光学刺激

双光子光遗传学已成为神经科学中不可或缺的技术，因为它能够精确和特定地操纵神经活动。通常采用无扫描全息方法来满足同时刺激神经集合的要求。然而，常用的圆盘图案无法实现单神经元分辨率，特别是在轴向维度上，并且它们固有的斑点降低了刺激效率。在这里，我们提出了一种用于体内神经元的高轴向分辨率光学刺激的新型无斑点串珠环图案. 使用染料池和荧光薄膜作为样品，我们验证了与圆盘图案相比，珠环图案可以实现更高的轴向分辨率和更好的定位能力。此外，我们在体内对小鼠 S1 皮层中的神经元进行了基于双光子的全光学生理学，并证明在刺激多个神经元时，与圆盘模式相比，通过串珠环模式获得的轴向分辨率可以提高 24% .