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: 8549309
• 负责人: JAY E STOCKLEY
• 金额: $ 22.34万
• 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/8549309
• 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; public health relevance; 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.

描述（由申请人提供）：精神病，包括精神分裂症，抑郁症和自闭症谱系障碍，尽管很明显它们主要代表皮质疾病，但仍然知之甚少。皮质是较高心理功能的主要部位，尽管进行了广泛的研究，但仍然没有关于皮质如何工作的统一理论。这是由于传统上依靠微电极来记录单个细胞活性的事实，这是因为神经科学家传统上依赖于微电极。但是，皮质电路由数百万个神经元组成，它 可以想象，仅单个细胞测量将不足以揭开大脑的功能。光学成像技术应对这种新兴的神经元电路活性水平，并能够在体外和体内成像神经元合奏的活性，同时保留单细胞分辨率，这是MRI或PET等脑成像技术（例如MRI或PET）。此外，遗传编码的光敏蛋白（光遗传学）和光化学（笼子）化合物的发展提供了机会，不仅可以对许多神经元的活性进行图像，还可以通过光学地控制它们。 尽管具有潜力，但当前的光学成像技术仍遭受了这样一个事实，即它们依靠激光器必须移动到每个像素以构建图像，从而使成像缓慢。此外，公共激光显微镜在2D中进行。为了解决这些问题，我们最近开发了一种新型的显微镜形式，该显微镜使用空间光调节剂（SLM）将激光束分成全息图案，可用于在3D中同时形象（或光激活）神经元。 SLM显微镜具有成为探索神经回路在脑部疾病中作用的理想方法的潜力。 Boulder非线性系统和哥伦比亚大学提议将其在建立SLM和SLM显微镜中的专业知识结合在一个两相项目中，最终的目标是使SLM显微镜成为神经科学和临床研究中的实践现实。在第一阶段，我们计划建立一个紧凑，廉价，用户友好的系统，该系统可以快速，3D成像和神经元的光活化。该设备将是自由对齐的，并与适当的软件集成，以便在几个神经生物学项目中使用它，包括成像完整的神经网络活动，对神经元点火的光学操纵，大脑连接性的功能映射，研究神经血管造影的功能映射，并研究了神经元素的神经元活动。在第二阶段，我们将扩展设计，以通过两光刺激来支持电生理记录，从而允许在活动物中进行3D成像和对皮质神经元的光刺激，例如醒着的表现啮齿动物的制剂。