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High Pressure Freezing and Freeze Substitution Maintain Structural Details and Protein Antigenicity in Protein-Storing Plant Cells

Published online by Cambridge University Press:  02 July 2020

J.E. Lonsdale ,
R.L. Jones  and
K.L. McDonald
J.E. Lonsdale
Affiliation:
Department of Plant and Microbial Biology, University of California, Berkeley, CA94720
R.L. Jones
Affiliation:
Department of Plant and Microbial Biology, University of California, Berkeley, CA94720
K.L. McDonald
Affiliation:
Electron Microscopy Laboratory, University of California, Berkeley, CA944720
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Extract

The cells of protein-storing plant tissues, including the aleurone of monocots and the cotyledons of dicots, have been widely used to study biological phenomena. The cells of protein-storing tissues share many characteristics including the presence of protein storage vacuoles (PSV) which act as reservoirs for the proteins, lipids and minerals required for germination. Many proteins synthesized in these tissues during germination are transported to the PSV. As a result, protein-storing tissues have become a model system for the study of protein transport.

Research in our laboratory focuses on the protein-storing barley aleurone layer. The aleurone layer is notoriously difficult to prepare for transmission electron microscopy (TEM). Aleurone cells are surrounded by thick cell walls that are difficult to penetrate with fixatives and resins. To increase penetration of chemicals, most of our studies are performed on protoplasts, aleurone cells that have had their walls enzymatically removed. Protoplasts, however, are sensitive to the osmolality of the fixation medium and solutions must be closely monitored during specimen preparation. The development of cells in the aleurone layer adds to the difficulty of preparation for TEM.

Type
Cryotechniques, Immunocytochemistry, and Electron Microscopy I. Molecular Approach
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 422 - 423
Copyright
Copyright © Microscopy Society of America

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References

1.Robards, AW and Sleytr, UB. In: Practical Methods in Electron Microscopy (1985).Google Scholar
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4. This research was supported by grants to R.L.J, from he National Science Foundation and the Competitive Research Grants Program of the USDA.Google Scholar