A facility for three-demensional computer reconstruction of subcellular structure is proposed. Thick (about .2-.5Mu) sections containing the biological structure are imaged using the Albany high-voltage electron microscope (HVEM) Biotechnology facility. Projections are obtained in a wide range of angles. Construction of a eucentric double tilt goniometer stage is proposed to replace the existing stage. The micrographs of tilt series are digitized, and the three-dimensional reconstruction is performed with a dedicated computer using the modified back projection algorithm. Windowing of the data, intermediate results and final reconstruction are viewed on a computer-linked high-speed graphics terminal. The facility will be used to study the detailed morphology of microtubular organizing centers and the three-dimensional organization of microtubules in reticulopods. A modular, user-friendly software system for image processing and three-dimensional reconstruction already exists. The general feasibility of this approach to subcellular structure visualization with the HVEM will be explored. The configuration and organization of a National resource facility allowing reconstruction from projections of thick sections will be defined, in terms of additional equipment needs and staffing.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Resource-Related Research Projects (R24)
Project #
3R24RR002033-03S1
Application #
3450564
Study Section
(SSS)
Project Start
1984-06-01
Project End
1987-12-31
Budget Start
1987-07-01
Budget End
1987-12-31
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost
Name
New York State Department of Health
Department
Type
DUNS #
002436061
City
Menands
State
NY
Country
United States
Zip Code
12204
Turner, J N; Valdre, U; Fukami, A (1989) Control of specimen orientation and environment. J Electron Microsc Tech 11:258-71
Turner, J N; Barnard, D P; Matuszek, G et al. (1988) High-precision tilt stage for the high-voltage electron microscope. Ultramicroscopy 26:337-43
McEwen, B F; Radermacher, M; Rieder, C L et al. (1986) Tomographic three-dimensional reconstruction of cilia ultrastructure from thick sections. Proc Natl Acad Sci U S A 83:9040-4