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Electron Tomographic Studies of a Large Macromolecular Assembly Preserved In Vitreous Ice: The Yeast Spindle Pole Body Core

Published online by Cambridge University Press:  02 July 2020

E. Bullitt
E. Bullitt*
Affiliation:
Boston University School of Medicine, Deptof Biophysics, 700 Albany Street, Boston, MA, 02118
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Extract

Advances in both electron cryomicroscopy and tomographic reconstruction methods have made it possible to collect and analyze data from large macromolecular assemblies frozen without fixation, sectioning, or staining. The Spindle Pole Body (SPB) is the microtubule organizing center in the yeast Saccharomyces, and is involved in mitosis, nuclear movement, budding, and mating. We have used electron cryomicroscopy and tomographic reconstruction to investigate the 1500Å diameter, 600Å high cylindrical core structure produced by limited heparin treatment of isolated SPBs.

Tilt series' of SPB cores preserved in vitreous ice were taken under low electron dose conditions. These images have been analyzed using iterative correlation alignment and computed back projections for tilt data without fiducial markers. In these studies, electron micrographs were digitized, and translationally aligned using correlation methods.

Type
Unique Approaches in Imaging, Computation and Communication for Characterization of the 3D Cell & Organelles I
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 434 - 435
Copyright
Copyright © Microscopy Society of America

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References

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5. This research was supported by grants from the National Science Foundation to Christopher W. Akey (Boston University School of Medicine) and EB.Google Scholar