High Speed Multiphoton Multifocal Whole Brain Imaging

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 Limited field of view due to aberrations induced by the intermediate optics 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 D at micron XY sampling and micron Z sampling in less than hours and will have a transformative effect on D histology and provide a crucial tool for researchers for a vast array of applications in neuroscience and other fields PROJECT NARRATIVE This Phase II application will build on our successful Phase I application where we proposed to build an ultra high speed Serial Two Photon STP Tomography microscope that can image en tire mouse brain and other organs with micron resolution in less than a day The microscope system will offer the highest imaging speed and sensitivity available for fluorescent subcellular whole organ imaging This proposal will have a broad impact on a range of biomedical applica tions in neuroscience cancer cardiac biology and pharmaceutical development