A novel 3D microscope for imaging and photostimulation

描述(申请人提供)：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，它们主要代表皮质障碍。大脑皮层是高级心理功能的主要场所，尽管进行了广泛的研究，但关于大脑皮层是如何工作的，仍然没有统一的理论。这是第一部分，因为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，大脑皮层回路是由数百万个神经元组成的，它 可以想象，单靠单细胞测量将不足以解开大脑的功能。光学成像技术解决了这种新兴水平的神经元电路活动，并能够在保持单细胞分辨率的同时，在体外和体内对神经元群的活动进行成像，这是MRI或PET等脑成像技术所无法做到的。此外，基因编码的光敏蛋白(光遗传学)和光化学(笼状)化合物的发展不仅为成像许多神经元的活动提供了机会，而且还提供了对它们进行光学控制的机会。尽管它们的潜力很大，但目前的光学成像技术存在一个问题，即它们依赖于必须移动到每个像素上的激光来构建图像，这使得成像速度变慢。此外，普通激光显微镜是在2D环境下进行的。为了取代这些问题，我们最近开发了一种新型的显微镜，它使用空间光调制器(SLM)，将激光分割成全息图案，可以用于同时在3D中成像(或光激活)神经元。SLM显微镜有可能成为探索神经回路在脑部疾病中作用的理想方法。博尔德非线性系统公司和哥伦比亚大学提议将他们在制造SLM和SLM显微镜方面的专业知识分两个阶段进行，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，我们计划建立一个紧凑、廉价、用户友好的系统，使神经元能够快速、3D成像和光激活。该设备将自我对准，并与适当的软件集成，以便它可以开箱即用，用于几个神经生物学项目中的应用，包括成像完整的神经网络活动、神经元放电的光学操作、大脑连接的功能映射、研究神经血管耦合，还可以用于在脑部疾病的动物模型中分析神经元活动。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，允许对活体动物的皮质神经元进行3D成像和光刺激，例如清醒行为的啮齿动物准备。

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.