High Speed Multiphoton Multifocal Whole Brain Imaging

高速多光子多焦全脑成像

• 批准号: 9358345
• 负责人: Timothy M. Ragan
• 金额: $ 67.13万
• 依托单位: TISSUEVISION, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9358345
• 关键词: Address; Agar; Air; Atlases; Awareness; Biology; Brain; Brain imaging; Cardiac; Collection; Color; Coupled; Custom; Data; Detection; Development; Electronics; Environment; Fiber; Histology; Hour; Image; Immersion Investigative Technique; Institutes; Liquid substance; Malignant Neoplasms; Maps; Microscope; Mus; Neurosciences; Optics; Organ; Pharmacologic Substance; Phase; Photobleaching; Photons; Platelet Factor 4; Price; Rattus; Research; Research Personnel; Resolution; Resources; Sampling; Scanning; Scheme; Science; Speed; System; Testing; Time; Tissue imaging; design; design and construction; experience; fluorescence imaging; imaging system; indexing; novel; residence; tomography; tool; two-photon

PROJECT SUMMARY/ABSTRACT In this application we describe the development of an ultra-high speed Serial Two-Photon (STP) Tomography microscope that can image entire organs with micron resolution in less than a day. It will offer the highest imaging speed and sensitivity available for fluorescent subcellular whole organ imaging. We will accomplish this by implementing a novel multi-foci multiphoton microscope (MMM) version of our current STP system that builds on TissueVision’s founding team’s extensive experience with MMM systems. It will address the two major problems with existing MMMs: 1. Limited field of view due to aberrations induced by the intermediate optics 2. Loss of emission photons in the descanned path. We will present details of a new excitation scheme that avoids aberrations and restricted field of views, and describe a non-descanned detection scheme that employs a high efficiency fiber coupled detection with an original interlaced scanning strategy that minimizes foci crosstalk. This proposal overcomes several long standing technical problems with MMM systems for deep tissue imaging, with a novel MMM design ideally suited for ex vivo whole organ imaging. The ability to fluorescently image a whole organ in 3D at 1 micron XY sampling and 2 micron Z sampling in less than 12 hours and will have a transformative effect on 3D histology and provide a crucial tool for researchers for a vast array of applications in neuroscience and other fields.

项目摘要/摘要 在此应用中，我们描述了超高速度串行两光子（STP）的发展 断层扫描显微镜可以在不到一天的时间内以微分分辨率成像整个器官。 它将提供可用于荧光亚细胞的最高成像速度和灵敏度 整个器官成像。 我们将通过实现新型的多焦点多光子显微镜（MMM）来实现这一目标 我们当前的STP系统的版本，该系统建立在Tissuevision的创始团队广泛的基础上 有MMM系统的经验。它将解决现有MMM的两个主要问题： 1。由于中间光学造成的畸变而导致的视场有限 2。在后路径中失去排放照片。 我们将介绍一种新的兴奋计划的详细信息，该计划避免了畸变和限制领域 视图，并描述采用高效率纤维的非研究的检测方案 将检测与原始的交织扫描策略相结合，从而最大程度地减少了焦点串扰。 该提案克服了MMM系统的几个长期技术问题，以进行深度 组织成像，具有新颖的MMM设计非常适合于体内整个器官成像。这 能够在1微米xy采样和2微米z中荧光在3D中成像整个器官 在不到12个小时的时间内进行采样，将对3D组织学产生变革性影响 为研究人员提供一个关键的工具，以在神经科学和其他 字段。