High Speed Multiphoton Multifocal Whole Brain Imaging

项目总结/摘要 在这个应用中，我们描述了超高速串行双光子（STP）的发展。 在不到一天的时间里，可以对整个器官进行微米级分辨率的成像。 它将提供最高的成像速度和灵敏度可用于荧光亚细胞 全器官成像 我们将通过实现一种新型的多焦点多光子显微镜（MMM）来实现这一目标 我们当前STP系统的一个版本，该版本建立在TissueVision创始团队的广泛 有MMM系统的经验。它将解决现有MMM的两个主要问题： 1.由于中间光学器件引起的像差导致的有限视场 2.去扫描路径中发射光子的损失。 我们将提出一个新的激发方案，避免像差和限制领域的细节。 视图，并描述了一个非去扫描检测方案，采用了高效率的光纤 耦合检测与原始隔行扫描策略，最大限度地减少焦点串扰。 该提议克服了用于深穿透的MMM系统的几个长期存在的技术问题。 组织成像，具有理想地适用于离体全器官成像的新型MMM设计。的 能够以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 在不到12小时内完成采样，将对3D组织学产生变革性影响， 为研究人员提供了一个重要的工具，用于神经科学和其他领域的大量应用。 领域的

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.