High Speed Multiphoton Multifocal Whole Brain Imaging

高速多光子多焦全脑成像

• 批准号: 8781939
• 负责人: Timothy M. Ragan
• 金额: $ 23.58万
• 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 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8781939
• 关键词: Address; Agar; Air; Atlases; Awareness; Biology; Brain; Brain Mapping; Brain imaging; Cancer Science; Cardiac; Collection; Color; Coupled; Data; Detection; Development; Environment; Fiber; Histology; Image; Institutes; Malignant Neoplasms; Marketing; Microscope; Mus; Neurosciences; Optics; Organ; Pharmacologic Substance; Phase; Photobleaching; Photons; Price; Rattus; Research; Research Personnel; Resolution; Sampling; Scanning; Scheme; Science; Source; Speed; System; Testing; Time; Tissues; design; experience; meetings; novel; public health relevance; residence; tomography; tool; two-photon

DESCRIPTION (provided by applicant): 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 a day 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.

描述（由申请人提供）：在本申请中，我们描述了一种超高速串行双光子（STP）断层扫描显微镜的开发，该显微镜可以在不到一天的时间内以微米分辨率对整个器官进行成像。它将为荧光亚细胞全器官成像提供最高的成像速度和灵敏度。我们将通过实施一种新的多焦点多光子显微镜（MMM）版本的我们目前的STP系统，建立在TissueVision的创始团队的丰富经验与MMM系统。它将解决现有MMM的两个主要问题：1。由于中间光学器件2引起的像差而导致的有限视场。在去扫描路径中发射光子的损失。我们将介绍一种新的激发方案的细节，该方案避免了像差和受限的视场，并描述了一种非去扫描检测方案，该方案采用了高效率的光纤耦合检测，并采用了原始的隔行扫描策略，使焦点串扰最小化。该提议克服了用于深部组织成像的MMM系统的几个长期存在的技术问题，具有理想地适合于离体整个器官成像的新颖的MMM设计。在不到一天的时间内以1微米XY采样和2微米Z采样对整个器官进行3D荧光成像的能力将对3D组织学产生变革性影响，并为研究人员在神经科学和其他领域的大量应用提供关键工具。