High-Resolution Cellular Imaging in Freely Moving Animals Using Fiber-Optic Multiphoton Fluorescence Endoscopy

该奖项支持光纤多光子内窥镜（FOME）的开发，该内窥镜能够在微米尺度上可视化自由移动动物组织内的细胞细节。该仪器将利用电信光纤和微光学的最新进展，允许用户在小到老鼠的动物身上安装或植入一个微光学探针。探针将使在行为测试室中自由移动的动物的细胞特性和动力学可视化。该仪器将使用多光子荧光激发来提供真正的三维成像，并在探针尖端位置的数百微米距离内进行薄光学切片。通过对活体动物行为和生理的宏观观察与对潜在细胞动力学的微观观察相结合的研究，FOME将有助于揭示生物体与其内部细胞之间的新关系。激发光和发射光将使用最先进的光纤在动物之间来回传输。通过注射或基因工程注入动物细胞的荧光标记可以选择性地标记特定细胞类别中的特定分子种类。用户将从计算机界面远程控制成像过程，而不必打断动物的行为。通过结合细胞和有机体水平的分析，该仪器将促进细胞生物学家、生理学家和行为科学家的合作研究。作为该项目的一部分，将建造两台辅助仪器，供受训人员和其他研究人员使用，以试点用于中央FOME成像站的新实验。这些辅助站将为生物科学中的现代光学成像技术以及神经元和神经回路的生物物理方面的新本科和研究生课程提供核心基础设施。

This award supports development of a fiber-optic multiphoton endoscope (FOME) capable of visualizing cellular details at the micron-scale within the tissues of freely moving animals. The instrument will exploit recent advances in telecom fiber- and micro-optics to allow the user to mount or implant a micro-optical probe on an animal as small as a mouse. The probe will enable visualization of cellular properties and dynamics in animals that are free to move within a behavioral testing chamber. The instrument will use multiphoton fluorescence excitation to provide true three-dimensional imaging with thin optical sectioning possible over distances of several hundred microns within the position of the probe tip. By enabling studies in which macroscopic observations of live animal behavior and physiology are concurrent with microscopic observations of underlying cellular dynamics, the FOME will help uncover new relationships between the organism and the cells within it. The excitation and emission light will be routed to and from the animal using state-of-the-art optical fibers. Fluorescent markers that are either injected or genetically engineered into the animal's cells will selectively tag particular molecular species within specific classes of cells. The user will control imaging sessions remotely from a computer interface, without having to interrupt animal behavior. By combining cellular- and organismic-level analysis, the instrument will facilitate collaborative studies by cellular biologists, physiologists and behavioral scientists. As part of the project, two auxiliary instrument will be constructed for use by trainees and other researchers to pilot new experiments destined for the central FOME imaging station. These auxiliary stations will provide core infrastructure for new undergraduate and graduate curricula on modern optical imaging techniques in the biological sciences and on biophysical aspects of neurons and neural circuits.