Holographic Optics Technology Core

双光子图案化光发生激发是一种新出现的扰动分布式神经的有力工具 具有细胞精确性和特异性的活性。然而，当前的双光子图案化技术 体内照射靶点有限的神经元具有相对较差的时间分辨率，尚未被 在大脑区域得到验证，目前还没有关于驱动哺乳动物行为反应的报道。这个 Technology Core将致力于一套技术先进但可行的传播和发展步骤， 提升团队的光学硬件、软件和光遗传学能力。这些进步将使 双光子光遗传刺激实现稳健精确的行为分布式神经控制 不同脑区和细胞类型的动物。所有三个研究项目都将与技术核心合作， 利用核心实验室在先进光学技术方面的独特专业知识，使我们能够 塑造激光波前，以单神经元分辨率询问大脑电路 啮齿动物的大脑。 首先，技术核心将整合和验证全光学成像和图案化的最佳实践 双光子微扰、实验管理软件和光敏探头。这些最新的- 艺术工具将在整个团队中传播。接下来，核心将推进和优化专用性和 通过探索新的光学、算法和探针定向解决方案，实现图案化双光子刺激的规模 为了提高刺激的特异性和稳健性，新的硬件和光学设计用于扩展 侧向和轴向可访问的刺激场，以及用于脑运动的闭环系统软件 补偿。最后，该核心将提高和优化图案化双光子的时间精度 使用速率优化的光遗传探针、空间光调制器(SLM)和高脉冲能量进行刺激 激光，并通过精确地将光学微扰与行为事件和本征事件同步 电动力学。 通过以前所未有的时间分辨率(低于10ms)刺激数百个神经元 通过研究不同脑区和细胞类型的动物，技术核心的工作将使这项研究成为可能 目的阐明大量神经元的协同活动是如何驱动行为的。优化 光学系统、软件系统和探头将成为以下结果比较的关键组件 感官模式。我们提案的一个独特优势是，技术团队参与执行 体内实验，确保在实验期间可以直接获得技术专业知识，并确保 问题的解决是由实验上相关的问题驱动的。

Two-photon patterned optogenetic excitation is a powerful emerging tool for perturbing distributed neural activity with cellular precision and specificity. However, current techniques for two-photon patterned illumination in vivo target a limited number of neurons with relatively poor temporal resolution, have not been validated across brain areas, and have not yet been reported to drive mammalian behavioral responses. The Technology Core will tackle a set of technically advanced yet feasible dissemination and development steps, advancing the team’s optics hardware, software, and optogenetics capabilities. These advances will enable two-photon optogenetic stimulation to achieve robust and precise distributed neural control in behaving animals across brain areas and cell types. All three research projects will work with the Technology Core, leveraging the unique expertise of the core laboratories in advanced optical technologies that allow us to sculpt laser wavefronts to interrogate brain circuits with single-neuron resolution in different sensory regions of the rodent brain. First, the Technology Core will integrate and validate the best practices for all-optical imaging and patterned two-photon perturbation, experimental management software, and light-sensitive probes. These state-of-the- art tools will be disseminated across the team. Next, the core will advance and optimize the specificity and scale of patterned two-photon stimulation, by exploring new optical, algorithmic, and probe-targeting solutions for improving stimulation specificity and robustness, new hardware and optical designs for extending the accessible stimulation field in both lateral and axial dimensions, and closed-loop software for brain motion compensation. Finally, the core will advance and optimize the temporal precision of patterned two-photon stimulation using rate-optimized optogenetic probes, spatial light modulators (SLMs), and high pulse-energy lasers, and by accurately synchronizing the optical perturbations with behavioral events and with intrinsic electrical dynamics. By enabling the stimulation of hundreds of neurons with unprecedented (below 10 ms) temporal resolution in behaving animals across brain areas and cell types, the work of the Technology Core will enable the research objective of elucidating how the cooperative activity of large groups of neurons drives behavior. Optimization of optical systems, software systems, and probes will be a critical component of result comparisons across sensory modalities. A unique strength of our proposal is that the technical teams are involved in performing the in vivo experiments, ensuring that technical expertise is directly available during experiments and that the problem solving is driven by experimentally relevant concerns.