Project Grant R44MH129132

THE DEVELOPMENT OF TABLETOP BRAIN-MAPPING PLATFORM - PROJECT SUMMARY CENTRAL NERVOUS SYSTEM (CNS) DISORDERS PLACE AN ENORMOUS BURDEN ON SOCIETY AND ARE AMONG THE MOST CHALLENGING AREAS FOR WHICH TO DEVELOP A THERAPEUTIC. A MAJOR REASON FOR THIS IS THAT THE BRAIN IS A COMPLEX, 3D, MULTISCALE ORGAN, WITH A HETEROGENOUS DISTRIBUTION OF COMPONENTS ACROSS HUNDREDS OF BRAIN REGIONS. CURRENTLY, EXTENSIVE BRAIN INITIATIVE EFFORTS ARE UNDERWAY TO MAP THIS COMPLEXITY BY CONDUCTING SURVEYS OF CONNECTIVITY PATTERN AND CELL TYPES. THE NEW FIELD OF 'SPATIAL BIOLOGY' IS PROVING TO BE A CENTRAL PLAYER IN THESE ENDEAVORS. WHILE CONTEMPORARY HISTOLOGICAL METHODS SUCH AS IHC ARE LIMITED TO IMAGING THREE OR FOUR PROTEINS, SPATIAL BIOLOGY APPROACHES CAN INVESTIGATE TENS TO THOUSANDS OF PROTEINS AND MRNA TRANSCRIPTS. UNFORTUNATELY, BARRIERS TO APPLYING SPATIAL BIOLOGY TECHNIQUES IN WHOLE 3D BRAINS ARE SUBSTANTIAL, INCLUDING THE HIGH COST OF INSTRUMENTATION, INCOMPATIBILITY WITH THICK TISSUES, AND EXTENSIVE DATA MANAGEMENT, ANALYSIS, AND VISUALIZATION CHALLENGES. EVEN OLDER, WELL- ESTABLISHED TISSUE PROTOCOLS ARE DIFFICULT TO APPLY ACROSS THE WHOLE BRAIN, AND SERIAL SECTION ANALYSIS WITH 2D SECTIONS IS FORMIDABLE OBSTACLE DUE TO ITS LABOR INTENSIVE AND ERROR PRONE NATURE. TOGETHER, THESE DIFFICULTIES PRESENT A SIGNIFICANT BARRIER TO APPLYING BOTH OLD AND NEW HISTOLOGICAL TECHNIQUES TO THE WHOLE BRAIN AND THUS HINDERS PROGRESS IN UNDERSTANDING THE CNS. THIS PROPOSAL WILL HELP ALLEVIATE THIS BARRIER BY BRINGING TO MARKET AN AFFORDABLE, TABLE-TOP HARDWARE AND SOFTWARE MOUSE BRAIN IMAGING AND TISSUE PROCESSING PLATFORM ACCESSIBLE TO THE NON-SPECIALIST NEUROSCIENCE LAB. IT IS BASED ON A MICROTOMY, SECTION CAPTURE, AND SLIDE MOUNTING TECHNOLOGY RECENTLY RELEASED IN TVI'S STATE-OF-THE-ART, MULTIPHOTON-BASED TISSUECYTE PLATFORM. THIS PROPOSAL WILL EXTEND THE CAPABILITIES OF THE TISSUECYTE PLATFORM TO SECTIONS AS THIN AS 10 MICRONS AND REPLACE THE TISSUECYTE'S ADVANCED MULTIPHOTON IMAGING WITH LESS EXPENSIVE BLOCK- FACE IMAGING. IN THIS PHASE II PROPOSAL WE WILL PACKAGE THIS NEW INTEGRATED TISSUE PROCESSING TECHNOLOGY INTO A CONVENIENT, AFFORDABLE TABLETOP INSTRUMENT. THIS GROUND-BREAKING DEVICE WILL COMBINE THE EASE OF WORKING FROM 2D SECTIONS WITH THE POWER OF 3D BRAIN ANALYSES AND HELP PROPEL FORWARD PROGRESS IN THE FIELD OF NEUROSCIENCE.