Adaptive optical microscopy for high-accuracy recording of neural activity in vivo

用于高精度记录体内神经活动的自适应光学显微镜

• 批准号: 10324548
• 负责人: NA Ji
• 金额: $ 57.74万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-15 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10324548
• 关键词: Afferent Neurons; Biological; Brain; Brain imaging; Detection; Dimensions; Event; Fluorescence; Fluorescence Microscopy; Functional Imaging; Goals; Hippocampus (Brain); Hypothalamic structure; Image; Lateral; Lead; Light; Lighting; Measurement; Membrane; Methodology; Methods; Microscopy; Modality; Monitor; Multiphoton Fluorescence Microscopy; Mus; Neurosciences; Optics; Penetration; Performance; Photons; Refractive Indices; Resolution; Sampling; Scanning; Speed; Spinal Cord; Structure; Subcellular structure; Synapses; Testing; Tissues; adaptive optics; base; brain tissue; combat; design; experience; imaging modality; in vivo; in vivo fluorescence; in vivo monitoring; lens; microendoscopy; millimeter; neural circuit; non-invasive imaging; prevent; relating to nervous system; temporal measurement; two-photon; voltage

PROJECT SUMMARY To understand the computations in the brain, we need to monitor the activity of neural circuits at high accuracy, which requires methodologies with high spatial and temporal resolution. Non-invasive and capable of resolving subcellular structures, optical microscopy has been extensively applied in the field of neuroscience, with a variety of methods developed to image neural activity at high speed, large depths, and/or over large spatial scales. For example, free-space angular-chirp-enhanced delay two-photon fluorescence microscopy was developed to record membrane voltage at kHz frame rate in the brain in vivo. Three-photon fluorescence microscopy, an emerging method that uses excitation light of longer wavelengths than two-photon fluorescence microscopy, has large penetration depths and is capable of imaging structures over 1-mm deep in the mouse brain. An alternative to the point-scanning multiphoton fluorescence microscopy above, single-photon widefield fluorescence microscopy has also been applied to in vivo monitoring of brain activity. Most commonly, the entire sample is illuminated and the emitted fluorescence collected by an objective lens and imaged with a camera, which enables fast activity imaging of superficial structures, sometimes over millimeters in lateral dimension. To obtain accurate measurements of neural activity in vivo, however, one has to combat the degradation of the resolving power of these microscopy methods when they are applied to brain tissue. The optical inhomogeneity of the biological tissue itself distorts the image-forming light and prevents all microscopy modalities from achieving their designed performance in vivo. When applied to activity imaging, such degradation can lead to erroneous conclusions. Here, we propose to optimize and apply adaptive optics methods developed in the Ji lab to select cutting-edge high- speed, large-depth, and large-scale activity recording modalities for high-accuracy measurements of neural activity in vivo.

项目摘要 要了解大脑的计算，我们需要以高精度监测神经回路的活性， 这需要具有高空间和时间分辨率的方法。无创和能够解决 亚细胞结构，光学显微镜已广泛应用于神经科学领域，多样 开发出用于高速，大深度和/或大型空间尺度上的神经活动的方法。为了 例如，开发了自由空间角增强延迟两光子荧光显微镜以记录 体内大脑中KHz帧速率的膜电压。三光子荧光显微镜，出现的 使用比两光子荧光显微镜的较长波长的激发光的方法具有较大的 渗透深度，能够在小鼠大脑深1毫米以上成像结构。替代 上面的扫描多光子荧光显微镜，单光子广场荧光显微镜 还应用于大脑活动的体内监测。最常见的是，整个样品被照亮了 以及通过客观镜头收集并用相机成像的发射荧光，这可以快速活动 浅表结构的成像，有时在横向尺寸的毫米上。获得准确 然而，在体内的神经活动的测量，必须对抗分辨能力的降解 这些显微镜方法将其应用于脑组织。生物学的光学不均匀性 组织本身会扭曲形成图像的光，并防止所有显微镜模式达到其设计 体内的性能。当应用于活动成像时，这种降解会导致错误的结论。这里， 我们建议优化和应用JI实验室中开发的自适应光学方法，以选择尖端的高级 - 速度，大深度和大规模的活动记录，用于高准确度测量的速度 体内活动。