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 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 实现为更强大和更精确的治疗方法打开了诱人的前景 重新调整异常电路活动。然而，我们仍然缺乏对神经活动模式的关键了解。 在正常行为期间以及这些模式在疾病中是如何改变的。人脑的许多啮齿动物模型 疾病现在是可用的，但神经学家仍然因为缺乏可视化活动的技术而受到阻碍 在表现良好的动物受试者的大脑中大量遗传识别的神经元中。Inspecix，Inc. 从斯坦福大学剥离出来，将一种微型荧光显微镜技术商业化 神经学家同时在清醒状态下可视化了多达1200个神经元中的钙动态，表现为啮齿类动物 细胞分辨率。显微镜可以很容易地携带在小鼠或大鼠的头上，并能够连续 从一只动物的同一组神经元记录几天到几周的时间。用于连词 凭借基因编码的钙离子指示器和我们定制设计的光学显微内窥镜探头， 显微镜能够瞄准预定义的神经元亚群，并进入无法进入的大脑区域 其他大规模记录技术。在第一阶段，我们开发并验证了新版本的 这款显微镜比斯坦福原型机的性能要高得多，也更结实。 围绕它，我们构建了nVista成像系统：一种用于活体脑成像的端到端解决方案，包括 微型显微镜，集成了高清摄像头和电子设备，并具有用户友好的数据采集功能 硬件和软件。NVista系统现已分发给100多个实验室进行Beta测试 环球网。这个第二阶段的项目将推动nVista系统在下一步的市场营销中取得进展 普通神经科学社区，通过以下目标。(1)创建下一代版本的 用于商业传播的nVista系统。利用来自早期采用者的反馈，我们将完善 系统的设计，通过结合电子调焦，提高了性能、可靠性和易用性 机制，实现更快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 包括一个新的底板连接机构，保护头安装锥体和电缆护套，光纤 探测器和支持换向器的光学数据链路；以及(3)开发下一代数据分析 具有更快的处理、可定制化和更好的可视化工具的平台。

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.