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

使用光纤多光子荧光内窥镜对自由移动的动物进行高分辨率细胞成像

• 批准号: 0352456
• 负责人: Mark Schnitzer
• 金额: $ 86.66万
• 依托单位: Stanford University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0352456&HistoricalAwards=false
• 关键词: Resolution; Cellular; Imaging; Freely; Moving

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.

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级显示自由运动动物组织中的细胞细节。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微型光学探头。该探测器将使动物的细胞属性和动力学可视化，这些动物可以在行为测试室中自由移动。该仪器将使用多光子荧光激发来提供真正的三维成像，并可以在探头尖端位置内数百微米的距离内进行薄光学切片。通过使活体动物行为和生理的宏观观察与潜在细胞动力学的微观观察同时进行的研究成为可能，Fome将有助于揭示有机体和其中的细胞之间的新关系。激发和发射的光将使用最先进的光纤来往于动物之间。被注射到动物细胞中或经过基因工程改造的荧光标记将选择性地标记特定类别细胞中的特定分子物种。用户将从计算机界面远程控制成像过程，而不必中断动物的行为。通过结合细胞和组织水平的分析，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，以引导新的实验，这些实验将运往中央Fome成像站。这些辅助站将为新的本科生和研究生课程提供核心基础设施，这些课程涉及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。