The proposed equipment would benefit at least nine R01 and one R21 NIH-funded projects carried out in the University of Kentucky. These projects cover wide range of health-related biomedical research, including studies on Alzheimer disease, viral infection, noise-induced hearing loss, spinal cord injury, hereditary forms of epilepsy, thrombosis, alcohol-induced neurotoxicity, chronic weakness, and diabetes. Collectively, these projects have generated more than 230 publications only in the last 5 years. The current application is in response to a significant need of NIH-funded Users to obtain excellent preservation of ultrastructural features in different cells/tissues without artifacts of fixation and dehydration. All these users explore intracellular or plasma membrane events that could be significantly compromised by chemical fixation and dehydration, which are inherent to ?classical? techniques of sample preparation for transmission electron microscopy (TEM). Therefore, the goal of this proposal is to acquire Leica high-pressure freezing (HPF) / freeze-substitution (FS) system. As of today, there is no single HPF system available either in Lexington campus or, in general, in the large neighboring region (~200 miles in radius) that includes Kentucky, Ohio, Vest Virginia, and the parts of Tennessee and Indiana. The proposed HPF/FS equipment will be installed and maintained in the Electron Microscopy Center (EMC) of the University of Kentucky, a core facility that operates as a recharge center. The PI on this application, Dr. Frolenkov, has a decades-long experience with various electron microscopy techniques, including HPF/FS techniques. The Director of EMC, Dr. Balk, has a decades-long experience in TEM and runs EMC since 2010. The everyday operation of the equipment will be supported by a skilled TEM technician. The operational cost of the equipment will be covered by moderate user fees. NIH-funded Major Users on this proposal will require ~80% of the accessible user time of the proposed equipment. The financial plan includes a commitment from the Vice President of Research of the University of Kentucky to cover 50% of the cost of the service contract. In addition, the Vice Dean of Research of the College of Medicine provides commitments for covering installation and initial start-up costs, as well as five-year back-up of the financial plan if service fees fall short of the anticipated costs of operating the system. Thus, the operation and maintenance of the proposed HPF/FS equipment are well supported. The proposed HPF/FS system will provide entirely new opportunities for the large group of NIH-funded researchers in the University of Kentucky, in one of the most hard-hit region of US economy, and help their numerous health-related projects.
The requested high-pressure freezing / freeze-substitution system will allow scientists in the University of Kentucky to prepare samples for electron microscopy observations without artifacts of chemical fixation and dehydration. This is very important for the range of NIH-funded health- related project carried out in the University of Kentucky, including studies on Alzheimer disease, viral infection, noise-induced hearing loss, spinal cord injury, hereditary forms of epilepsy, thrombosis, alcohol-induced neurotoxicity, chronic weakness, and diabetes.
