Building and sharing next generation open-source, wireless, multichannel miniaturized microscopes for imaging activity in freely behaving mice

 描述（由申请人提供）：神经科学中最大的挑战之一是了解大脑中的神经回路如何处理，编码，存储和检索信息。要应对这一挑战，需要有方法来记录行为自由的动物的完整神经网络的活动。随着神经活动的遗传编码指标和光遗传致动器的发展，现在需要在长时间内对自由移动的小鼠中大量已识别神经元的活动进行成像和操纵的方法。需要多通道成像来根据其独特的基因表达谱或投影模式明确识别单个神经元，并且需要一个灵活的平台，以便可以轻松调整范围以解决各种神经科学问题。光遗传学的能力需要在细胞活动模式和行为之间建立因果关系。最后，目前可用的技术是有限的，因为它需要用电线拴着老鼠，限制了它们的行为范围和与其他动物或环境互动的能力。此外，商业小型化示波器极其昂贵，并且不能改变以满足个别最终用户的需求。在这里，我们寻求通过开发真正开源的下一代双通道光遗传学功能，无线，小型化显微镜来弥补现有技术的不足，用于成像和跟踪自由移动小鼠中大型神经细胞群的活动模式。我们将设计，制造，优化和测试：用于成像两个荧光团的双通道显微镜（Aim 1），用于成像和光遗传学激发的光遗传学显微镜（Aim 2），无线小型化显微镜（Aim 3），以及具有组合双通道，光遗传学和无线功能的显微镜（Aim 4）。在整个过程中，我们将建立一个在线环境，与神经科学界分享我们的显微镜，为其他人建立，修改和植入显微镜并使用我们的软件分析神经活动数据解除障碍。我们的新型可穿戴显微镜将对神经科学产生变革性的影响，首次允许对数百个已识别的神经元和其他细胞（如自由行为动物中的星形胶质细胞）的活动进行成像和操作。

 DESCRIPTION (provided by applicant): One of the biggest challenges in neuroscience is to understand how neural circuits in the brain process, encode, store, and retrieve information. Meeting this challenge will require methods to record the activity of intact neural networks in freely behaving animals. Spectacular advances with the development of genetically encoded indicators of neural activity and optogenetic actuators now call for methods to image and manipulate the activity of large populations of identified neurons in freely moving mice over prolonged periods of time. Multi-channel imaging is needed to unequivocally identify individual neurons based on their unique gene expression profiles or projection patterns, and a flexible platform is needed so that the scopes can be easily adapted to address diverse neuroscience questions. Optogenetic capability is needed to draw causal connections between cellular activity patterns and behavior. Finally, currently available technology is limited because it requires the mice to be tethered by wires, limiting their range of behaviors and ability to interact with other animals or their environment. In addition, commercial miniaturized scopes are extremely expensive, and cannot be altered to meet individual end-user needs. Here, we seek to remedy shortfalls in existing technology by developing truly open source next-generation two-channel optogenetics-capable, wireless, miniaturized microscopes for imaging and tracking activity patterns of large neural-cell populations in freely moving mice. We will design, manufacture, optimize and test: a two-channel microscope for imaging two fluorophores (Aim 1), an optogenetics-capable microscope for imaging and optogenetic excitation (Aim 2), a wireless miniaturized microscope (Aim 3), and a microscope with combined two-channel, optogenetics, and wireless capability (Aim 4). All throughout, we will build an online environment for sharing our microscopes with the neuroscience community, lifting barriers for others to build, modify, and implant the microscopes and analyze neural activity data with our software. Our new wearable microscopes will have a transformative impact on neuroscience by permitting for the first time the imaging and manipulation of the activity of hundreds of identified neurons and other cells such as astrocytes in freely behaving animals.