NCS-FO: Imaging synaptic activity deep in the brain using super-resolution cannula microscopy

提案# 1533611研究所：犹他大学标题：NCS-FO：使用超分辨率插管显微镜成像大脑深处的突触活动目的：本项目将开发一种工具，用于自由运动动物突触的高分辨率（100纳米）成像，用于神经元研究。它将通过开发和整合紧凑和轻量级的试管显微镜，体外荧光成像以及成像突触的相关技术和方法来实现这一目标。该项目的长期愿景是使用廉价的技术对自由移动的老鼠的大脑深处进行高分辨率成像，从而阐明信息处理和记忆的基本基础。特定突触强度的变化被认为是记忆编码和存储的基础，然而，由于技术上的限制，在完整的大脑中，这一理论的实验证据很少，因为视觉化涉及经验依赖学习的特定突触模式。这个项目旨在克服这一限制，将一个简单、廉价的插管改造成超分辨率荧光显微镜。这项技术的商业化将在基础科学和工程得到广泛传播之后进行。技术：本提案的目的是利用计算插管显微镜（CM）和新的突触活动分子报告器，以低于100纳米的分辨率成像大脑深处的神经元活动，神经元结构和蛋白质定位。CM将允许以前所未有的空间分辨率对清醒、自由活动的动物的大脑进行成像。目前用于自由活动动物的技术仅限于近表面的大脑成像，包括带有活动部件的大而重的头部阶段，并且无法在不严重损害周围组织的情况下深入大脑。这项提议的最终目标是允许对自由活动的动物的单个突触进行成像。我们已经开发了使用CM进行体外荧光成像的框架。在本项目中，我们将对CM进行扩展，使其能够：(1)超分辨率（100nm分辨率）荧光显微镜和(2)深度（1mm）脑深部成像，可以看到自由运动动物脑深部单个突触的成像活动和蛋白质定位。个体突触强度的变化被认为是大脑学习和记忆的基础，然而这一大脑功能的基本理论缺乏实际的实验证据来支持它。我们的项目将使研究能够解决在单个突触中介导行为和信息处理的分子事件的因果作用。

Proposal # 1533611Institute: University of UtahTitle: NCS-FO: Imaging synaptic activity deep in the brain using super-resolution cannula microscopyObjective: This project will develop a tool for high-resolution (100-nm) imaging of synapses in freely moving animals for neuronal studies. It will accomplish this goal by the development and integration of compact and lightweight cannula microscopy with in vitro fluorescence imaging with accompanying technology and methodologies for imaging synapses.Non-Technical The long-term vision of this project is to image with high resolution deep inside the brain of freely moving mice using inexpensive technologies so as to elucidate the fundamental basis of information processing and memory. Changes in synaptic strength at specific synapses are thought to underlie memory encoding and storage, yet there is very little experimental evidence for this theory in the intact brain due to technical limitations of visualizing the specific synaptic pattern involved in experience-dependent learning. This project aims to overcome this limitation by transforming a simple, inexpensive cannula into a super-resolution fluorescence microscope. Commercialization of this technology will be pursued after the fundamental science and engineering has been demonstrated for widespread dissemination. Technical:The objective of this proposal is to image neuronal activity, neuron structure and protein localization deep in the brain with sub-100nm resolution using computational cannula microscopy (CM) and novel molecular reporters of synaptic activity. CM will allow imaging of the brain in awake, freely moving animals at unprecedented spatial resolution. Current techniques in freely moving animals are limited to imaging the brain near the surface, include large and heavy head stages with moving parts, and cannot penetrate deep into the brain without significant damage to surrounding tissue. The ultimate goal of this proposal is to allow imaging of individual synapses in freely moving animals. We have already developed the framework for in vitro fluorescence imaging using CM. During this project, we will extend CM to enable: (1) super-resolution ( 100nm resolution) fluorescence microscopy and (2) deep-brain imaging (depth 1mm) with the vision of imaging activity and protein localization in individual synapses in the deep brain of freely moving animals. Changes in the strength of individual synapses are thought to underlie learning and memory in the brain, yet this fundamental theory of brain function lacks tangible experimental evidence to support it in vivo. Our project will enable studies that address the causal role of molecular events at individual synapses in mediating behavior and information processing.