PFI:AIR - TT: Proof-of-concept fiber-based miniature multiphoton microscope using adaptable optics

这个PFI: AIR技术翻译项目的重点是将微型自适应光学与多光子激光扫描显微镜相结合，以满足对多功能、可植入的三维成像系统的需求，用于研究清醒行为动物的大脑活动。这种纤维耦合微型多光子显微镜非常重要，因为它使神经科学家能够前所未有地研究动物在进行不同行为任务时复杂神经回路的动力学。该项目将导致我们的产品在自由移动的动物操作的概念验证演示。这种光纤耦合的微型多光子显微镜可以通过主动聚焦实现全三维扫描，并且没有机械运动部件，设计紧凑，重量轻。该产品可以很容易地连接到商用激光扫描显微镜。这些特点可以降低成本，灵活配置波长或成像方式，并且与市场上领先的头戴式显微镜系统相比，可以提供更大脑容量的成像。该项目解决了以下技术差距，因为它从研究发现转化为商业应用。尽管光学显微镜在生物系统研究方面取得了迅速进展，但一个主要的差距是能够使用这些先进的成像工具对自由运动动物的大脑进行动态研究。我们提出在紧凑的光纤耦合设计中结合双光子激发和深度扫描的优点。目前市售的微型显微镜只允许单个成像平面，在成像过程中不允许主动聚焦，也不允许目标成像和光遗传刺激。微型多光子光纤耦合显微镜将利用光纤束将激光扫描信号传递到远端显微镜头，并利用微型自适应光学元件实现快速主动聚焦。这将实现高通用性的应用，如快速感兴趣区域成像、光遗传刺激和多光子技术的深度成像。参与该项目的人员，包括本科生和研究生，将通过与商业伙伴的会议、参加技术会议和参加外联活动获得技术翻译的经验。该项目与3i（智能成像创新）合作，这是一家专业提供先进光学显微镜系统的国际公司。这次合作将使这种光纤耦合显微镜进入商业系统，并加快向全球商业市场的交付。

This PFI: AIR Technology Translation project focuses on combining miniature adaptable optics with multiphoton laser scanning microscopy to fill the need for a versatile, implantable, three-dimensional imaging system for studying brain activity in awake behaving animals. The fiber-coupled miniature multiphoton microscope is important because it allows neuroscientists an unprecedented ability to study dynamics of complex neuronal circuits while animals are undergoing different behavior tasks. The project will result in a proof-of-concept demonstration of our product operating in freely moving animals. The fiber-coupled miniature multiphoton microscope will allow full three-dimensional scanning through active focusing with no mechanically moving parts in a compact, light weight design. The product can be readily connected to a commercial-laser scanning microscope. These features allow for reduced cost, flexibility in configuration of the wavelength or imaging modality to be performed, and can provide imaging over larger brain volumes when compared to the leading head-mounted microscope systems in this market space.This project addresses the following technology gaps as it translates from research discovery toward commercial application. Although light microscopy has rapidly advanced for studies of biological systems, one major gap is the ability to use these advanced imaging tools for dynamic studies in the brain in freely moving animals. We propose to combine the advantages of two-photon excitation with depth scanning using adaptive optics in a compact fiber-coupled design. Current commercially available miniature microscopes only allow for single imaging planes, do not enable active focusing during imaging and do not allow both targeted imaging and optogenetic stimulation. The miniature multiphoton fiber-coupled microscope will use a fiber-bundle to translate the laser-scanned signal to the distal microscope head and miniature adaptable optical elements to provide fast active focusing. This will accomplish high versatility for applications such as fast region-of-interest imaging, optogenetic stimulation, and depth imaging with multiphoton techniques. Personnel involved in this project including undergraduate and graduate students, will receive experience in technology translation through meetings with business partners, attending technology conferences, and participating in outreach events.This project engages 3i (Intelligent Imaging Innovations), an international company that specializes in delivering advanced optical microscope systems. This collaboration will allow implementation of this fiber-coupled microscope into commercial systems and speed delivery to a global commercial market.