Miniature, Integrated Fluorescence Microscopes for In Vivo Brain Imaging

描述(由申请人提供)：今天，神经回路在神经精神疾病中的作用越来越受到重视。然而，我们仍然缺乏关于神经活动的正常模式以及这些模式在疾病中如何出错的关键知识。尽管大脑研究人员已经建立了许多人类大脑疾病的小鼠模型，但目前还没有技术可以可视化表现良好的小鼠大脑中大量单个、遗传识别类型的神经元的活动-理想情况下是在多只小鼠中并行进行。 获得如此大规模数据集的能力对于识别大脑疾病的神经生理学特征非常重要，也是开发重新调整异常活动模式的治疗手段的先决条件。荧光显微镜在追踪神经活动方面具有关键优势。然而，尽管传统的荧光显微镜提供了成像大脑细胞动力学所需的时空分辨率，但它们既不允许在行为自由的小鼠身上进行研究，也不能扩展到对大量动物受试者的研究。如果荧光显微镜可以变得更小、更便携、更便宜，那么原则上就可以对大量行为正常的小鼠进行平行研究。Inscope公司已经从斯坦福大学剥离出来，将微型集成荧光显微镜商业化--这是一种成像技术，可以帮助神经科学家可视化清醒行为的小鼠和大鼠的神经回路动力学。斯坦福大学的原型显微镜已经能够成像小脑微循环，并允许可视化数百个单个神经元内的钙动态(在一些实验中需要数周时间)，因为动物以自然的方式自由活动。最近，《自然》、《麻省理工学院技术评论》和几家媒体对核心微型集成显微镜技术创新及其在研究大脑及其疾病方面的前景进行了专题报道。在第一阶段，Inscope ix的目标是开发和测试一套新的原型显微镜，这种显微镜性能要高得多，坚固耐用，是用户友好的端到端解决方案的一部分，用于自由行为啮齿动物的活体脑成像。具体地说，我们将：(1)设计和创造我们的小型化、集成化显微镜的新版本。我们将进一步开发核心技术，并纳入几项改进，以显著提高成像性能，并扩展活体脑成像的能力，包括：(A)实现空间成像 (B)开发一种数字高速旋转换向器，使其能够对大脑活动进行无监督的成像研究；(C)制造一种坚固可靠的显微镜外壳，适合大批量低成本制造。(2)开发数据采集和处理的配套硬件和软件。我们将创建一个紧凑和用户友好的USB兼容盒，用于图像采集和显微镜控制，以及易于使用的图形用户界面(GUI)。(3)制作并测试10台带外围设备的新型微型显微镜。我们将制造并内部测试我们的新设计，然后将10个原型分发给精心挑选的Beta实验室进行体内测试和验证。到第一阶段结束时，我们预计将收到来自测试版实验室的大量体内使用反馈，为第二阶段批量生产和推出市场就绪的产品奠定基础。

DESCRIPTION (provided by applicant): There is a rising emphasis today on the role of neural circuitry in neuropsychiatric disease. However we still lack crucial knowledge of both normal patterns of neural activity and how these patterns go awry in disease. Although brain researchers have already created mouse models of many human brain diseases, presently there is no technology that can visualize the activity of large numbers of individual, neurons of genetically identified types in the brains of behaving mice - ideally in multiple mice in parallel. The capacity to obtain such large-scale data sets is important towards identifying neurophysiologic signatures of brain disease and is a prerequisite for developing therapeutic means of re-tuning aberrant activity patterns. Fluorescence microscopy has key advantages for tracking neural activity. However, while conventional fluorescence microscopes offer the spatiotemporal resolution needed for imaging the brain's cellular dynamics, they neither permit studies in freely behaving mice nor are scalable for studies of large numbers of animal subjects. If fluorescence microscopes could be made small, portable, and cheap, then in principle large numbers of behaving mice could be studied in parallel. Inscopix, Inc. has spun-out of Stanford University to commercialize miniature, integrated fluorescence microscopes - imaging technology that helps neuroscientists visualize neural circuit dynamics in awake behaving mice and rats. Prototype microscopes at Stanford are already enabling imaging of cerebellar microcirculation and permitting visualization of Ca2+ dynamics within hundreds of individual neurons (over weeks in some experiments) as the animal behaves freely in a naturalistic manner. The core miniature, integrated microscope technological innovation and its promise for studying the brain and its diseases was recently featured in Nature, MIT Technology Review, and several media outlets. In Phase I Inscopix aims to develop and test a new set of prototype microscopes that are significantly higher-performing, robust and part of a user-friendly end-to-end solution for in vivo brain imaging in freely behaving rodents. Specifically, we will: (1) Desig and create a new version of our miniaturized, integrated microscope. We will further develop the core technology and incorporate several improvements to significantly enhance imaging performance and extend the capabilities for in vivo brain imaging, including: (a) Attaining spatial resolution finer than 1 ¿m over fields-of-view up to 1 mm2; (b) Developing a digital, high-speed rotary commutator enabling unsupervised, imaging studies of brain activity; (c) Creating a robust and reliable microscope housing suitable for low-cost manufacturing in large volumes. (2) Develop accompanying hardware and software for data acquisition and processing. We will create a compact and user-friendly USB-compatible box for image acquisition and microscope control along with an easy-to-use Graphical User Interface (GUI). (3) Fabricate and test 10 new miniature microscopes with accompanying peripherals. We will fabricate and internally test our new designs before distributing 10 prototypes to carefully chosen beta labs for in vivo testing and validation. By the end of Phase I we expect to have received considerable in vivo usage feedback from beta labs, laying the foundation for volume production and roll-out of a market-ready product in Phase II.