Using the existing facility for tomographic three-dimensional reconstruction and the Albany high voltage electron microscope (HVEM) Biotechnology Resource, a number of in house projects involving structures related to cell motility and mitosis (cilia basal body, centriole, kinetochore) will be investigated at a resolution approaching 5 nm. In addition, outside collaborations will be pursued on systems that pose difficult questions concerning shape and 3-D connectivity of structural components. In HVEM tomography, thick sections of plastic embedded biological material are tilted with a precision goniometer. The projections collected on the photographic film re scanned and combined by the computer into a three-dimensional representation of the object. The facility will be further developed to simplify and increase the efficiency of scanning, alignment and data processing, and to achieve the highest, most isotropic resolution possible. To this end, conical data collection, the use of Boolean object modelling, and various restoration schemes will be explored.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM040165-05
Application #
3297535
Study Section
Special Emphasis Panel (SSS (C))
Project Start
1988-01-01
Project End
1993-12-31
Budget Start
1992-01-01
Budget End
1993-12-31
Support Year
5
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Wadsworth Center
Department
Type
DUNS #
110521739
City
Menands
State
NY
Country
United States
Zip Code
12204
Liu, Y; Penczek, P A; McEwen, B F et al. (1995) A marker-free alignment method for electron tomography. Ultramicroscopy 58:393-402
Landis, W J (1995) Tomographic imaging of collagen-mineral interaction: implications for osteogenesis imperfecta. Connect Tissue Res 31:287-90
McEwen, B F; Arena, J T; Frank, J et al. (1993) Structure of the colcemid-treated PtK1 kinetochore outer plate as determined by high voltage electron microscopic tomography. J Cell Biol 120:301-12
Wilson, C J; Mastronarde, D N; McEwen, B et al. (1992) Measurement of neuronal surface area using high-voltage electron microscope tomography. Neuroimage 1:11-22
Landis, W J; Hodgens, K J; McKee, M D et al. (1992) Extracellular vesicles of calcifying turkey leg tendon characterized by immunocytochemistry and high voltage electron microscopic tomography and 3-D graphic image reconstruction. Bone Miner 17:237-41
Barnard, D P; Turner, J N; Frank, J et al. (1992) A 360 degrees single-axis tilt stage for the high-voltage electron microscope. J Microsc 167:39-48
Woodcock, C L; McEwen, B F; Frank, J (1991) Ultrastructure of chromatin. II. Three-dimensional reconstruction of isolated fibers. J Cell Sci 99 ( Pt 1):107-14
McEwen, B F; Song, M J; Landis, W J (1991) Quantitative determination of the mineral distribution in different collagen zones of calcifying tendon using high voltage electron microscopic tomography. J Comput Assist Microsc 3:201-10
Frank, J (1989) Three-dimensional imaging techniques in electron microscopy. Biotechniques 7:164-73