Request for a ThermoFisher Helios 5UC DualBeam

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7. Project Summary/Abstract: This request is for funds to purchase a ThermoFisher Helios 5UC DualBeam Focused Ion Beam Scanning Electron Microscope (FIB-SEM) system. This instrument is being requested by a group of NIMH-funded investigators who have a significant need to acquire 3-dimensional (3D) volumes of brain tissue imaged at nanometer resolution. The requested FIB-SEM system will be housed in a dedicated facility within the Translational Neuroscience Program (TNP) at the University of Pittsburgh School of Medicine (UPSOM). Installation of the requested Helios 5 UC DualBeam FIB-SEM represents an important expansion of existing shared microscopy facilities that are specialized for, and dedicated to, brain tissue imaging. There is a pressing and prevalent need for 3D large volume electron microscopy (VEM) approaches that permit studies interrogating the basis of brain mechanisms underlying complex behaviors and their disruptions in psychiatric disorders. The advent of 3D volume electron microscopy (VEM) addresses this critical need by generating highly detailed connectomic maps of individually resolved synapses, and the sub-synaptic organelles within, across densely packed, large volumes of brain tissue. The requested FIB-SEM system permits automated and quantitative VEM imaging of brain tissue that avoids many of the limitations associated with other VEM approaches. Although FIB-SEM application to neuroscience research is relatively recent, we show that this technology can be applied to postmortem human brain tissue to create 3D, information-rich datasets of volumes of brain tissue neuropil ready for quantitative analysis. Thus, with this technology we can interrogate the fine structure of the nervous system and pinpoint structural disruptions contributing to brain dysfunction. The need for this instrument is also demonstrated by the overwhelming response from NIMH-funded researchers in the UPSOM. We clearly explain how the requested FIB-SEM will advance the research objectives of current NIMH-funded projects and how this resource is invaluable for generating new NIMH grant applications. The investigators in this proposal have decades of experience in traditional EM approaches, but these methods cannot generate the quantity, or the quality, of data generated by FIB-SEM imaging. Therefore, there is a single prime reason for this application: to provide critically needed, cutting-edge, high quality automated VEM imaging to NIMH supported users within the UPSOM.