A novel 3D microscope for imaging and photostimulation

用于成像和光刺激的新型 3D 显微镜

• 批准号: 8455342
• 负责人: JAY E STOCKLEY
• 金额: $ 37.35万
• 依托单位: BOULDER NONLINEAR SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-21 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8455342
• 关键词: Address; Affect; Animal Model; Animals; Area; Award; Biological Assay; Biological Neural Networks; Brain; Brain Diseases; Brain Mapping; Brain imaging; Calcium; Cells; Clinical Research; Collaborations; Computer software; Coupling; Development; Devices; Diagnostic; Disease; Etiology; Functional disorder; Goals; Image; Imaging Techniques; In Vitro; Individual; Laboratory Research; Laser Microscopy; Lasers; Legal patent; Letters; Life; Light; Location; Long-Term Effects; Magnetic Resonance Imaging; Malignant Neoplasms; Marketing; Measurement; Measures; Mental Depression; Methods; Microelectrodes; Microscope; Microscopic; Microscopy; Molecular; Monitor; Movement; Neurobiology; Neurons; Neurosciences; Neurosciences Research; Optical Methods; Optics; Outcome; Pattern; Phase; Photochemotherapy; Photons; Population; Positron-Emission Tomography; Preparation; Problem Solving; Proteins; Psyche structure; Research; Research Technics; Resolution; Rodent; Role; Sampling; Scanning; Schizophrenia; Site; Solutions; Source; Speed; Surface; System; Techniques; Technology; Therapeutic; Three-Dimensional Imaging; Universities; Work; autism spectrum disorder; awake; base; design; in vivo; neural circuit; novel; novel strategies; optical imaging; optogenetics; palliative; photoactivation; prototype; response; stem; theories; three dimensional structure; tool; two-dimensional; two-photon; user-friendly

DESCRIPTION (provided by applicant): Mental disease, including schizophrenia, depression and autism spectrum disorders, are still poorly understood, although it is clear that they mostly represent cortical disorders. The cortex is the primary site of higher mental functions, and despite extensive research, there is still no unified theory of how the cortex works. This is partl due to the fact that neuroscientists have traditionally relied on microelectrodes to record the activity of individual cells. However, cortical circuits are composed of millions of neurons and it is conceivable that single cell measurements alone will not be sufficient to unravel function of the brain. Optical imaging techniques tackle this emergent level of neuronal circuit activity and enable to image the activity of neuronal ensembles, in vitro and in vivo, while preserving single cell resolution, something that brain imaging techniques such as MRI or PET, cannot do. Moreover, the development of genetically encoded photosensitive proteins (optogenetics) and optochemical (caged) compounds offers the opportunity to not only image the activity of many neurons but also to optically control them. In spite of their potential, current optical imaging techniques suffer form the fact that they rely on lasers which have to be moved to each pixel to build an image, making the imaging slow. Moreover, common laser microscopy is performed in 2D. To supersede those problems, we have recently developed a novel form of microscopy that uses spatial light modulators (SLM), to split the laser beam into a holographic pattern that can be used to image (or photoactivate) neurons simultaneously in 3D. SLM microscopy has the potential of becoming the ideal method with which to explore the role of neural circuits in brain diseases. Boulder Nonlinear Systems and Columbia University propose to combine their expertise in building SLMs and in SLM microscopy in a two-phase project with the ultimate goal of making SLM microscopy a practical reality in neuroscience and clinical research. In the first phase we plan to build a compact, inexpensive, user- friendly system that enables fast, 3D imaging and photoactivation of neurons. The device will be self-aligning and integrated with appropriate software so that it can be used, out of the box, for applications in several neurobiological projects including imaging intact neural network activity, optical manipulation of neuronal firing, functional mapping of brain connectivity, investigating neurovascular coupling, and also be used for assaying neuronal activity in animal models of brain disease. In Phase II we will extend the design to support electrophysiological recording with two-photon excitation, allowing 3D imaging and photostimulation of cortical neurons in living animals, such as awake behaving rodent preparations. PUBLIC HEALTH RELEVANCE: Microscopy with spatial light modulators (SLMs) enables use of optical techniques to study neuronal circuit activity, to both monitor and manipulate the activity of neuronal ensembles, in vitro and in vivo. Boulder Nonlinear Systems and Columbia University propose the development of a compact, inexpensive, user-friendly SLM based microscope ("Pocketscope") that enables fast, 3D imaging and photoactivation of neurons. The device will find widespread use in neuroscience research including imaging intact neural network activity, optical manipulation of neuronal firing, functional mapping of brain connectivity investigating neurovascular coupling, and also be used for assaying neuronal activity in animal models of brain disease.

描述（由申请人提供）：人们对精神疾病，包括精神分裂症、抑郁症和自闭症谱系障碍仍然知之甚少，尽管很明显它们主要代表皮质疾病。皮层是高级心理功能的主要场所，尽管进行了广泛的研究，但仍然没有关于皮层如何工作的统一理论。这部分是由于神经科学家传统上依靠微电极来记录单个细胞的活动。然而，皮质回路由数百万个神经元组成，它 可以想象，仅单细胞测量不足以揭示大脑的功能。光学成像技术解决了神经元回路活动的这种新兴水平，并能够在体外和体内对神经元群的活动进行成像，同时保留单细胞分辨率，这是 MRI 或 PET 等大脑成像技术无法做到的。此外，基因编码的光敏蛋白（光遗传学）和光化学（笼式）化合物的发展不仅为许多神经元的活动成像提供了机会，而且还为光学控制它们提供了机会。 尽管具有潜力，但当前的光学成像技术仍受到以下事实的困扰：它们依赖于激光，激光必须移动到每个像素才能构建图像，从而使得成像速度变慢。此外，常见的激光显微镜是在 2D 中进行的。为了解决这些问题，我们最近开发了一种新型显微镜，它使用空间光调制器 (SLM)，将激光束分成全息图案，可用于同时对神经元进行 3D 成像（或光激活）。 SLM 显微镜有可能成为探索神经回路在脑部疾病中的作用的理想方法。 博尔德非线性系统公司和哥伦比亚大学提议在一个两阶段项目中结合他们在构建 SLM 和 SLM 显微镜方面的专业知识，最终目标是使 SLM 显微镜在神经科学和临床研究中成为现实。在第一阶段，我们计划构建一个紧凑、廉价、用户友好的系统，能够实现神经元的快速 3D 成像和光激活。该设备将具有自对准功能，并与适当的软件集成，因此可以开箱即用地用于多个神经生物学项目的应用，包括对完整神经网络活动进行成像、神经元放电的光学操纵、大脑连接的功能图谱、研究神经血管耦合，并且还可用于分析脑部疾病动物模型中的神经元活动。在第二阶段，我们将扩展设计以支持双光子激发的电生理记录，从而允许对活体动物（例如清醒行为啮齿动物制剂）的皮层神经元进行 3D 成像和光刺激。 公共健康相关性：带有空间光调制器 (SLM) 的显微镜可以利用光学技术来研究神经元回路活动，从而在体外和体内监测和操纵神经元群的活动。 Boulder Nonlinear Systems 和哥伦比亚大学提议开发一种紧凑、廉价、用户友好的基于 SLM 的显微镜（“Pocketscope”），可实现神经元的快速 3D 成像和光激活。该设备将在神经科学研究中得到广泛应用，包括对完整神经网络活动进行成像、神经元放电的光学操纵、研究神经血管耦合的大脑连接功能映射，并且还可用于分析脑部疾病动物模型中的神经元活动。