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Use of Freeze Substitution for TEM Studies of Fungal Spores

Published online by Cambridge University Press:  02 July 2020

C. W. Minis  and
E. A. Richardson
C. W. Minis
Affiliation:
Department of Plant Pathology, University of Georgia, Athens, GA30602
E. A. Richardson
Affiliation:
Department of Botany, University of Georgia, Athens, GA30602
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Extract

Since its initial use to fix fungal hyphae (1), plunge freezing followed by freeze substitution has become the “gold standard” for TEM studies of fungal hyphae and spores. In this presentation we discuss results we have obtained using plunge freezing and freeze substitution to fix various types of spores produced by plant pathogenic fungi. Examples include basidiospores and aeciospores of rust fungi, teliospores of rust and smut fungi and conidia of a variety of ascomycetes and deuteromycetes.

Generally speaking, plunge freezing followed by freeze substitution usually yields best results when spore diameter does not exceed 10-15 μm. However, in a number of cases we (2) have obtained good to excellent results with much larger spores. A key to good freezing appears to be direct exposure of spores to liquid propane during plunging. We have had best results by placing spores on small pieces of dialysis membrane and then plunging these membranes. In most cases the spores remain attached to membranes throughout the freezing, substitution, infiltration and embedment procedures.

Type
Light and Electron Microscopic Techniques for the Study of Plant Pathogenic Fungi and Their Interactions with Host Plants
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 684 - 685
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Howard, R. J. and Aist, J. R., J. Ultra. Res., 66 (1979)224CrossRefGoogle Scholar
2.Mims, C. W. and Richardson, E. A., Newsletter Southeastern Micro. Soc., 15 (1999)1Google Scholar