Hostname: page-component-cd9895bd7-jkksz Total loading time: 0 Render date: 2024-12-28T15:44:24.402Z Has data issue: false hasContentIssue false

Use of low temperature scanning electron microscopy to observe frozen hydrated seed tissues of Glycine max

Published online by Cambridge University Press:  19 September 2008

Robert W. Yaklich*
Affiliation:
Soybean and Alfalfa Research Laboratory, USDA, ARS, Bldg. 008, BARC-West, Beltsville, MD 20705, U.S.A.
William P. Wergin
Affiliation:
Electron Microscopy Laboratory, USDA, ARS, Bldg. 177B, BARC-East, Beltsville, MD 20705, USA
Eric F. Erbe
Affiliation:
Electron Microscopy Laboratory, USDA, ARS, Bldg. 177B, BARC-East, Beltsville, MD 20705, USA
*
*Correspondence

Abstract

Evidence is provided to show that frozen, hydrated seeds and seed tissues can be observed by low temperature field emission scanning electron microscopy. This technique allows preparation and observation of seed samples that contain 13–60% water without altering their moisture content. The technique provides information about the surface structure of seeds and also allows specimens to be fractured to reveal internal features of tissues. Futhermore, both halves of fractured specimens can be retained, examined and photographed either as two-dimensional micrographs or as stereo pairs when three-dimensional observation (stereology) or quantitative measurements (stereometry) are desired. Use of this technique avoids artefacts associated with chemical fixation, dehydration, and critical point drying—procedures that are usually required to prepare seed tissues for scanning electron microscope examination. This technique does not affect the degree of hydration in specimens; it can be used to localize water in tissues to determine their degree of hydration. This technique should find wide application in developmental studies of seeds, especially during maturation, imbibition and germination.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 1994

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Burris, J.S. (1973) Effect of seed maturation and plant population on soybean seed quality. Agronomy Journal 65, 440441.CrossRefGoogle Scholar
Beckett, A. and Read, N.D. (1986) Low-temperature scanning electron microscopy. pp 4580in Aldrich, H.C. and Todds, W.J. (Eds) Ultrastructural techniques for microorganisms. New YorkPlenum.CrossRefGoogle Scholar
Echlin, P., Paden, R., Dronzek, B. and Wayte, R. (1970) Scanning electron microscopy of labile biological material maintained under controlled conditions. Scanning Electron Microscopy 1970, 4956.Google Scholar
Fehr, W.R., Caviness, C.E., Burmood, D.T. and Pennington, J.S. (1971) Stage of development descriptions for soybeans, Glycine max L. Merr. Crop Science 11, 929931.Google Scholar
ISTA (1985) International rules for seed testing. Rules. Seed Science and Technology 13, 299355.Google Scholar
McDonald, M.B. Jr., Vertucci, C.W. and Roos, E.E. (1988) Soybean seed imbibition: water absorption by seed parts. Crop Science 28, 993997.CrossRefGoogle Scholar
Read, N.D. (1991) Low-temperature scanning electron microscopy of fungi and fungus-plant interactions. pp 1729in Mendgen, K. and Lesemann, D.E. (Eds) Electron microscopy of plant pathogens. Berlin, Springer-Verlag.Google Scholar
Read, N.D. and Jeffree, C.E. (1991) Low-temperature scanning electron microscopy in biology. Journal of Microscopy 161, 5972.CrossRefGoogle ScholarPubMed
Wergin, W.P. and Erbe, E.F. (1989) Increasing the versatility of an EM scope SP2000A Sputter Cryo System on a Hitachi S-570 scanning electron microscope. Scanning 11, 293303.Google Scholar
Wergin, W.P. and Erbe, E.F. (1991a) Increasing resolution and versatility in low temperature conventional and field emission scanning electron microscopy. Scanning Microscopy 5, 927936.Google Scholar
Wergin, W.P. and Erbe, E.F. (1991b) Using high vacuum evaporation to obtain high resolution low temperature images of freeze-fractured membranes from yeast. Proceedings of the 49th Annual Meeting of the Electron Microscope Society of America 49, 514515.Google Scholar
Wergin, W.P. and Erbe, E.F. (1991c) Introduction to the advantages and problems associated with low temperatures scanning electron microscopy. Scanning 13, 2426.Google Scholar
Wergin, W.P. and Erbe, E.F. (1992a) Techniques for obtaining and observing complementary images with a lowtemperature field emission SEM and subsequent comparison of the identical cells in freeze-etch replicas viewed with a TEM. Scanning 14, 1730.Google Scholar
Wergin, W. P. and Erbe, E.F. (1992b) Recent advancements in low temperature scanning electron microscopy. Scanning 14, 11401142.Google Scholar
Wergin, W.P. and Erbe, E.F. (1992c) Advantages of complementary stereo images as viewed with the low temperature field emission scanning electron microscope. Proceedings of the 50th Annual Meeting of the Electron Microscope Society of America 50, 13141315.Google Scholar
Wergin, W.P. and Pooley, C. (1988) Photographic and interpretive artifacts. pp 175204in Crang, R.F.E. and Klomparens, K.L. (Eds) Artifacts in biological electron microscopy Chapter 9. New York, Plenum Publishing Corporation.Google Scholar
Wolf, W.J., Baker, F.L. and Bernard, R.L. (1981) Soybean seed-coat structural features: pits, deposits, and cracks. Scanning Electron Microscopy 3, 531544.Google Scholar
Yaklich, R.W., Kulik, M.M. and Garrison, C.S. (1979) Evaluation of vigor in soybean seeds: influence of date of planting and soil type on emergence, stand, and yield. Crop Science 19, 242246.Google Scholar
Yaklich, R.W., Vigil, E.L. and Wergin, W.P. (1986) Pore development and seed coat permeability in soybean. Crop Science 26, 616624.CrossRefGoogle Scholar
Yaklich, R.W., Vigil, E.L., Erbe, E. F. and Wergin, W.P. (1992) The fine structure of aleurone cells in the soybean seed coat. Protoplasma 167, 108119.Google Scholar