Funds are requested for the purchase of a large format CCD camera to enhance 3-D electron microscopy at Florida State University. The camera will be mounted on a Philips/FEI CM300-FEG and replace an existing 2k x 2k CCD camera which will be transferred to a CM120. Both microscopes and both CCD cameras are housed in a shared university facility, called the Biological Science Imaging Resource (BSIR), which contains instrumentation for both light and electron microscopy used by many researchers on campus and in the north Florida area. The BSIR is professionally staffed by employees that are paid by the university, not by user fees, thereby providing substantial support for users who only pay fees to cover service contracts and expendable supplies. NIH funded projects that will be enhanced by this equipment include the following: electron tomography of Ascaris sperm motility complexes, SIV and HIV viruses, insect flight muscle, cell adhesion complexes and adeno-associated virus. In addition the camera will find use in various cryoEM 3-D imaging projects that require high throughput data collection. These include adeno-associated virus in complex with various receptors, heavy meromyosin in many different states, membrane proteins from Mycobacterium tuberculosis, and 2-D arrays of various kinds formed on lipid monolayers. The new instrumentation will allow for development of algorithms for increasing the area that can be reconstructed by electron tomography which may need corrections for focus gradient and for imaging distortions.
| 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 |
