The Biological Science Imaging Resource (BSIR) at Florida State University (FSU) is the focal point for cryoelectron microscopy and 3-D electron microscopy in the state of Florida. No other University facility has a comparable emphasis or comparable facilities. This application requests funds to alleviate a critical need for equipment dedicated to the preparation of thin vitreous ice films and for specimen holders dedicated to electron tomography. This equipment will support the research of the major BSIR users, Drs. Kenneth Taylor, Kenneth Roux and Thomas Roberts and visitors from the lab of a long standing collaborator, Dr. Michael Chapman, who recently moved to Oregon Health & Science University. Funds are requested for 4 main items of equipment including (1) Vitrobot computer controlled specimen freezer for preparing thin vitrified suspensions, (2) a plasma cleaner for producing hydrophilic holey carbon films, (3) a high tilt cryoelectron tomography specimen holder and (4) a dual axis, high tilt, room temperature specimen holder. The study of thin, vitreous, frozen hydrated suspensions is a large part of the cryoEM research emphasis at FSU, but preparation of these specimens is currently a bottleneck in a number of projects. This bottleneck will be removed by purchase of the combination plasma cleaner for making hydrophilic support films and Vitrobot for preparing the thin vitrified aqueous films. Electron tomography is a major emphasis of three applicant groups while the fourth is beginning to explore the possibilities. The dual axis tomography holder will improve both throughput and quality of reconstructions obtained from rapidly frozen, freeze substituted, plastic embedded biological specimens while the cryotomography holder will enhance 3-D imaging of frozen hydrated specimens. These items will support NIH funded research in (1) the fundamental mechanism of force generation in a myogenic muscle, which has health relevence to cardiac muscle function; (2) smooth muscle myosin regulation, which has health relevance in hypertension; (3) adhesion structures and migrating cells, which has health relevence in cancer; (4) the structure of HIV and SIV viruses which are fundamental to understanding AIDS; and (5) the development of Adeno Associated Virus as a gene therapy vector. ? ? ?
|O'Donnell, Jason; Taylor, Kenneth A; Chapman, Michael S (2009) Adeno-associated virus-2 and its primary cellular receptor--Cryo-EM structure of a heparin complex. Virology 385:434-43|