A multi-plane 3-photon microscope for volume imaging in NHP cortex

用于 NHP 皮层体积成像的多平面 3 光子显微镜

• 批准号: 10205806
• 负责人: Jack Waters
• 金额: $ 80.17万
• 依托单位: ALLEN INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10205806
• 关键词: Animals; Behavior; Brain; Cell physiology; Cells; Computer software; Cortical Column; Development; Disease; Electron Microscopy; Experimental Designs; Fluorescence Microscopy; Future; Goals; Health; Human; Image; Imaging Techniques; Institutes; Macaca; Measurement; Measures; Microscope; Microscopy; Motion; Mus; Neocortex; Neurons; Optics; Patients; Performance; Photons; Population; Primates; Property; Protocols documentation; Research; Resolution; Rodent; Role; Sensory; Structure; Techniques; Technology; Testing; Tissues; Universities; Visual; Visual Cortex; Washington; area striata; brain tissue; color processing; design; experimental study; fluorescence microscope; insight; microscopic imaging; nonhuman primate; novel strategies; three photon microscopy; tool; two-photon; visual information; visual processing; web site

PROJECT ABSTRACT Recent advances in microscopy permit volume imaging - the near-simultaneous imaging of many neurons in a circuit - with 2-photon (2P) excitation, yielding new insights into the circuits underlying relatively simple behaviors. Volume imaging techniques and 2P excitation have been limited mainly to small animals such as rodents, including studies of mouse cortex. Mouse cortex differs from neocortex in non-human primates and humans and it is unlikely that studies of mouse cortex will be sufficient to understand the role of cortex in humans. In this R34 application, we propose to build a multi-plane 3-photon (3P) fluorescence microscope to perform volume imaging in macaques. Unlike 2P excitation, 3P excitation supports imaging deep into macaque cortex. In a future R01 application, we will use this 3P microscope to image large volumes of macaque primary visual cortex, studying how motion information is pooled across neurons in layer 4B and pairing 3P microscopy with serial-section electron microscopy to study the processing of color information, particularly in layer 4Cβ. We will pursue four specific aims: - Specific aim 1: Construct and test a multi-plane 3P microscope - Specific aim 2: Measure visual tuning of ~50,000 neurons in layers 1-4 in a macaque. - Specific aim 3: Extend multi-plane 3P imaging through layers 5 and 6 of macaque cortex. - Specific aim 4: Make hardware designs, software and protocols freely available.

项目摘要 显微镜的最新进展允许体积成像-电路中许多神经元的几乎同时成像- 与2光子（2 P）激发，产生新的见解电路相对简单的行为。体积 成像技术和2 P激发主要局限于小动物，如啮齿动物，包括研究 小鼠皮层小鼠皮层不同于非人类灵长类动物和人类的新皮层， 小鼠大脑皮层的研究将足以理解人类大脑皮层的作用。 在这个R34应用中，我们建议建立一个多平面3光子（3 P）荧光显微镜来进行体积测量。 在猕猴中成像。与2 P激发不同，3 P激发支持深入猕猴皮层的成像。未来R 01 应用程序，我们将使用这种3 P显微镜对猕猴初级视觉皮层的大量图像进行成像，研究如何 运动信息汇集在4 B层的神经元之间，并将3 P显微镜与连续切片电子显微镜配对 显微镜来研究颜色信息的处理，特别是在层4Cβ中。 我们将追求四个具体目标： - 具体目标1：构建并测试多平面3 P显微镜 - 具体目标2：测量猕猴1-4层中约50，000个神经元的视觉调谐。 - 具体目标3：将多平面3 P成像扩展到猕猴皮质的第5层和第6层。 - 具体目标4：免费提供硬件设计、软件和协议。