ClearScope

Combined in vivo and ex vivo three dimensionalDwhole brain imaging of non transgenic and transgenic animal models holds the promise of novel insights into neural network connectivity patternsWith regard to ex vivo light microscopic imaging ofD whole brain datasetsthe best approach is brain clearing followed by whole brain light sheet microscopyLSMbecause of its unique combination of speedD resolving powerand low phototoxicity compared to confocal and multiphoton microscopyUnlike other methodsthe combined brain clearingLSM approach makes it possible to use intact tissue and retain all intracellular connections within the brain structureHoweverLSM systems commercially available are not suitable for ex vivo light microscopic imaging ofD whole brain datasets in advanced connectomics researchRecentlyDrRaju TomerDeptBiolSciColumbia UnivNew YorkNYdeveloped light sheet theta microscopyLSTMessentially a unique arrangement of two light sheets oblique to the specimen and one detection objective perpendicular to the specimenThis novel microscope is the basis for a number of capabilities in LSTM that are not all available with any other commercially available LSM systemsThe LSTM technology has distinct advantages over confocal and other light sheet microscopesincluding the unmatched ability to image thicker tissue specimens over a larger lateral areaXYat higher optical resolutionswhile maintaining fast imaging speedhigh imaging qualityand low photo bleachingThis promising technology serves as the basis for this Lab to Marketplace proposal to develop the ClearScopewhich refines and improves DrTomerandapos s original LSTM system to create a successful commercial microscope for wide spread adoptionThe key technical objectives for developing the ClearScope as a commercial product include creating and testingia ClearScope prototype based on an optimized microscope hardware designiinovel microscope hardware components for the ClearScopecomprising a novel chamber that contains the investigated specimen and the immersion mediumand a novel detection objective changeriiinovel control and image acquisition software for the ClearScopeand importantlyivnovel software that surpasses the existing state of the art technology to assemble acquired image stacks into largeD image volumes exceedingTB without need to downsample the image informationThe production version of the ClearScope will benefit the neuroscience research communitypharmacological and biotechnological Randamp Dand society in general by improving understanding of neural network connectivity patterns as well as the neuropathological underpinnings of the large scale connectional alterations associated with human neuropsychiatric and neurological conditionsIn particularthis will result in an improved basis for developing novel treatment strategies for complex brain diseases The proposed project will enable important new researchthat is not currently feasibleinto whole brain neural network connectivity patterns by means of ex vivo light microscopic imaging of three dimensional whole brain datasetsTo achieve thisthe proposed project will create the ClearScope light sheet microscope featuring signficant capabilities to image thick specimens of unlimited lateralXYsize with resolution that permits investigatons of subcellular structures to distinguish adjacent neuronal processesaxonsdendritesvaricosities and dendritic spinesBecause these features are not all available with any commercially available light sheet microscopesthe benefit for the neuroscience research communitypharmacological and biotechnological Randamp Dand society in general will be the possibility to better understand neural network connectivity patterns as well as the neuropathological underpinnings of the large scale connectional alterations associated with human neuropsychiatric and neurological conditionsultimately resulting in an improved basis for developing novel treatment strategies for complex brain diseases