Miniature, integrated and mass-producible fluorescence microscopes for in vivo brain imaging in freely behaving rodents

项目摘要 越来越多的人意识到，异常的神经回路活动是许多疾病的核心表现， 神经精神疾病，包括自闭症、精神分裂症、抑郁症和阿尔茨海默病。这 这一认识为基于以下的更强大和精确的治疗方法开辟了诱人的前景： 异常电路活动的重新调谐。然而，我们仍然缺乏对神经活动模式的关键理解， 以及这些模式在疾病中如何改变。许多人类大脑的啮齿动物模型 疾病现在是可用的，但神经科学家仍然受到缺乏可视化活动技术的阻碍 在行为动物的大脑中大量的基因识别的神经元中。Insopix，Inc. 从斯坦福大学分离出来，将一种微型荧光显微镜技术商业化， 神经科学家在清醒的行为啮齿动物中同时观察多达1200个神经元中的Ca2+动力学， 细胞分辨率显微镜很容易携带在小鼠或大鼠的头上，并且能够连续观察。 在一只动物的同一组神经元中记录数天至数周的时间。结合使用 通过基因编码的Ca2+指示剂和我们定制设计的光学显微内窥镜探头， 显微镜能够靶向预定义的神经元亚群，并进入无法进入的大脑区域。 其他大型录音技术。在第一阶段，我们开发并验证了新版本的 该显微镜比最初的斯坦福大学原型具有更高的性能和更强大的性能。 围绕它，我们构建了nVista成像系统：一个用于体内脑成像的端到端解决方案，包括 微型显微镜，集成高清摄像头和电子设备，以及用户友好的数据采集 硬件与软件nVista系统现已分发到100多个实验室进行beta测试， 地球仪。该第二阶段项目将推动nVista系统的下一步营销， 通过以下目标，一般神经科学社区。(1)创建下一代版本的 用于商业传播的nVista系统。利用早期采用者的反馈，我们将改进 系统的设计，更大的性能，可靠性和易用性，通过纳入电子聚焦 机制，实现更高速度的数据采集，并与其他数据采集无缝对接 系统;（2）通过开发nVista显微镜的附件阵列将该技术扩展到大鼠， 包括一个新的基板连接机制，保护头戴式锥体和电缆护套， 探测器和光学数据链，以支持制冷机;（3）开发下一代数据分析 该平台具有更快的处理速度、可定制性和更好的可视化工具。

Project Summary There is increasing awareness that aberrant neural circuit activity is a core manifestation of many neuropsychiatric diseases, including autism, schizophrenia, depression and Alzheimer's disease. This realization has opened tantalizing prospects for more powerful and precise therapeutic approaches based on retuning of abnormal circuit activity. However, we still lack crucial understanding of neural activity patterns during normal behavior and how these patterns are altered in disease. Many rodent models of human brain diseases are now available, but neuroscientists are still hindered by lack of a technology for visualizing activity in large populations of genetically identified neurons in the brains of behaving animal subjects. Inscopix, Inc. spun out of Stanford University to commercialize a miniature fluorescence microscope technology that lets neuroscientists visualize Ca2+ dynamics in up to 1200 neurons simultaneously in awake, behaving rodents at cellular resolution. The microscope is easily carried on the head of a mouse or rat, and enables continuous recording from the same group of neurons in a single animal for periods of days to weeks. Used in conjunction with genetically encoded Ca2+ indicators and our custom-designed optical microendoscope probes, the microscope enables targeting of predefined neuronal subpopulations, and entry to brain regions inaccessible to other large-scale recording technologies. In Phase I, we developed and validated a new version of the microscope that is substantially higher performing and more robust than the original Stanford prototype. Around it we built the nVista imaging system: an end-to-end solution for in vivo brain imaging that includes the miniature microscope with an integrated HD camera and electronics, and user-friendly data acquisition hardware and software. The nVista system has now been disseminated for beta testing to over 100 labs around the globe. This Phase II project will move the nVista system forward in its next steps towards marketing to the general neuroscience community, through the following aims. (1) Create a next-generation version of the nVista system for commercial dissemination. Making use of feedback from early adopters, we will refine the system's design for greater performance, reliability and ease of use, by incorporating an electronic focusing mechanism, enabling higher speed data acquisition and seamless interfacing with other data collection systems; (2) Extend the technology to rats by developing an accessory array for the nVista microscope that includes a new base plate attachment mechanism, protective head-mounted cone and cable sheathing, optical probes, and an optical data link to support commutators; and (3) Develop a next-generation data analysis platform with faster processing, customizability, and better visualization tools.