FAST 3D DIMENSIONAL IMAGING OF NEURONAL ACTIVITY AND PLASTICITY

描述(申请人提供)：神经科学的一个中心目标是了解动物和人类的行为是如何由大脑的回路产生的。实现这一目标的进展在一定程度上取决于记录神经元群体活动的能力。为了从本地电路进行密集记录，最广泛使用的技术之一是钙成像。双光子显微镜的钙成像目前允许研究人员同时记录数百个神经元；虽然与过去的方法相比有了重大改进，但它仍然只占几乎任何局部电路中神经元总数的一小部分。在我看来，要全面理解神经元回路是如何驱动行为的，最重要的技术障碍是很难确定哪些神经元首先计算一个特定的“决定”或活动模式。最近，我们开发了一种称为目标耦合平面照明(OCPI)显微镜的方法，用于对整个组织体积进行快速荧光成像。它不是一次从一个像素收集数据，而是使用一张薄薄的光片一次收集整个图像。OCPI显微镜使我们能够以高速和信噪比同时记录大约一万个神经元。在我们关于嗅觉系统的科学工作中，OCPI显微镜揭示了感觉外周检测到的东西，化学刺激是如何编码的，大脑电路的空间排列，甚至不同的人是如何感知世界的。在这里，我建议扩大OCPI显微镜的适用范围。在目标1中，我们将开发新的方法来更深入地了解活着的大脑。在目标2中，我们将开发程序，根据神经元的生理特性对单个神经元进行“标记”。在目标3中，我们将开发和共享算法来处理由OCPI显微镜产生的TB大小的数据集。这些目标将使OCPI显微镜能够解决新的问题，并在神经科学界广泛传播。

DESCRIPTION (provided by applicant): A central goal of neuroscience is to understand how animal and human behaviors are generated by the circuits of the brain. Progress towards this goal has relied in part on the ability to record the activity of neuronal populations. To record densely from a local circuit, one of the most widely-used techniques is calcium imaging. Calcium imaging by two-photon microscopy currently allows investigators to record from hundreds of neurons simultaneously; while a major improvement over past approaches, it is nevertheless a small fraction of the total number of neurons in almost any local circuit. The difficulty of determining which neurons first compute a particular "decision" or pattern of activity is, in my view, the most important technical barrier to a comprehensive understanding of how neuronal circuits drive behavior. Recently, we developed a method, called Objective-Coupled Planar Illumination (OCPI) microscopy, to perform fast fluorescence imaging of whole tissue volumes. Instead of collecting data from one pixel at a time, it uses a thin sheet of light to collect entir images at once. OCPI microscopy has allowed us to record from approximately ten thousand neurons simultaneously at high speeds and signal-to-noise ratio. For our scientific work on the olfactory system, OCPI microscopy has revealed what the sensory periphery detects, how chemical stimuli are encoded, the spatial arrangement of circuits in the brain, and even how different individuals perceive the world. Here I propose to extend the domain of applicability of OCPI microscopy. In aim 1, we will develop new methods to see deeper into the living brain. In aim 2, we will develop procedures to "tag" individual neurons based on their physiological properties. In aim 3, we will develop and share algorithms to process the terabyte-sized datasets produced by OCPI microscopy. These aims will allow OCPI microscopy to address new questions and to be disseminated widely throughout the neuroscience community.