Acquisition of a Leica high pressure freezer for the cryo-based electron microscopy techniques - PROJECT SUMMARY / ABSTRACT Funds are requested to purchase a Leica EM ICE high-pressure freezer to prepare electron microscopy samples for projects focused around mental health research within the Vanderbilt Brain Institute (VBI), a multi-disciplinary research institute comprised of faculty at Vanderbilt University (VU) and Vanderbilt University Medical Center (VUMC). The requested instrument will be integrated into and managed by the Vanderbilt Cell Imaging Shared Resource (CISR), a microscopy core with a 20+ year history of successful microscopy management, training, and acquisition of new technology. This instrument capable of fast-freezing live or fixed samples in vitreous ice for downstream applications including freeze-substitution TEM, cryo-FIBSEM, and cryo-ET. This equipment is essential to progress the research of the 7 Major Users within Vanderbilt and the VBI with projects surrounding neuronal cell biology, synapse assembly and plasticity, synaptic signaling, glutamate receptors, and communication between cell populations. These molecular and cellular investigations will advance already funded NIMH and NIH projects totaling over $4M in FY23 in the areas of psychiatric disorders, Alzheimer’s, ALS, learning and memory, Rett syndrome (RTT), childhood neurodegenerative diseases and more. The proposed high-pressure freezing system is based on the needs of the VBI and Major Users to capture neuronal processes with the highest resolution and fidelity possible to understand fundamental advance these NIMH and NIH funded projects. The Leica EM ICE high-pressure freezer includes: 1) The base unit high- pressure freezing that uses pressurized liquid nitrogen to both generate pressure and cool samples simultaneously; 2) Electric and light stimulation modules to capture synaptic events at the point of freezing 3) A cell and tissue incubator system to load live samples into the EM ICE; 4) Sapphire based freezing modules to allow correlative light and electron microscopy either before or after freezing, and conventional flat carriers for freeze-substitution and freeze-fracture/etch applications. This system would be housed in renovated electron microscopy suite in Medical Center North which is directly adjacent to the VBI and the majority of the User labs. Vanderbilt University is providing financial support to promote the long-term operation of the instrument. If awarded, the Office of the Dean of Basic Sciences is committed to providing $75,896 for CISR service contracts, staff salary, computer purchases and upgrades, and reduction in user fees over the first 5 years. Technical support and training of new users will be facilitated by the exceptional staff within the CISR, which has a greater than 20-year history of assisting over 300 labs on campus in both sample preparation and imaging. Moreover, this equipment will be housed in newly renovated electron microscopy space that is centrally located near the Major User group. Collectively, the proposed high-pressure freezing system, together with outstanding institutional and core facility support, will enable the long-term objective of supporting and advancing neuroscience and mental health research Vanderbilt.
