All holographic two-photon electrophysiology

全全息双光子电生理学

• 批准号: 10616937
• 负责人: Hillel Adesnik
• 金额: $ 407.69万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10616937
• 关键词: 3-Dimensional; Acceleration; Action Potentials; Address; Algorithms; Animals; BRAIN initiative; Benchmarking; Brain; Calibration; Cell Size; Code; Cognition; Development; Disease; Electrophysiology (science); Engineering; Esthesia; Functional Imaging; Health; Heating; Holography; Image; Lasers; Light; Lighting; Measures; Mediating; Membrane; Microscope; Microscopy; Monitor; Morphologic artifacts; Motion; Neurons; Neurosciences Research; Opsin; Optics; Performance; Population; Process; Resolution; Sampling; Scanning; Shapes; Source; Speed; Spottings; System; Technology; Testing; Time; Update; Writing; computer generated; imaging approach; imaging platform; in vivo; light scattering; microbial; microscopic imaging; millisecond; neural; neural network; neural patterning; neuroimaging; neuroregulation; new technology; novel; novel strategies; optogenetics; redshift; scale up; sensor; spatiotemporal; temporal measurement; two-photon; voltage; yeast two hybrid system

PROJECT SUMMARY Achieving a detailed understanding of the neural codes of sensation, action, and cognition will require technologies that can both sample and perturb neural activity with millisecond precision and cellular resolution across large populations of neurons. We will develop an all-optical holographic two-photon microscope that can simultaneously record and perturb population neural activity with cellular resolution and millisecond precision. To achieve this, we will leverage multispectral temporally focused three-dimensional (3D) wavefront shaping. This new form of hybrid two-photon functional imaging will enable simultaneous illumination of user-defined ensembles of neurons in 3D with cell-size spots of two-photon excitation light at two different wavelengths – one for imaging a voltage sensor and one for controlling neural activity optogenetically. The two holographic optical paths will be completely independent and update at >kilohertz speed with the ability to sample and perturb the membrane voltage of tens to hundreds of neurons at a time. In tandem, we will develop task-optimized two-photon excitable red-shifted genetically encoded voltage sensors (GEVIs) and ultrapotent blue-shifted microbial opsins that are spectrally separable. The development of an all-holographic read/write microscope will permit neuronal recording and perturbation on spatial and temporal scales that are currently well beyond reach, which could substantially facilitate systems neuroscience research.

项目摘要 要想详细了解感觉、动作和认知的神经编码，就需要能够在大量神经元中以毫秒级精度和细胞分辨率对神经活动进行采样和扰动的技术。我们将开发一种全光全息双光子显微镜，它可以同时记录和干扰群体神经活动，具有细胞分辨率和毫秒级精度。为了实现这一点，我们将利用多光谱时间聚焦三维（3D）波前整形。这种新形式的混合双光子功能成像将使用户定义的神经元集合在3D中与两种不同波长的双光子激发光的细胞大小的斑点同时照明-一种用于成像电压传感器，另一种用于控制神经活动光遗传学。这两条全息光路将是完全独立的，并以>千赫兹的速度更新，能够一次对数十到数百个神经元的膜电压进行采样和扰动。同时，我们将开发任务优化的双光子可激发的红移遗传编码电压传感器（GEVI）和光谱可分离的超高效蓝移微生物视蛋白。全全息读/写显微镜的发展将允许神经元记录和扰动的空间和时间尺度上，目前远远超出了范围，这可以大大促进系统神经科学研究。