Back to Search
Start Over
An improved method for affinity probe localization in whole cells of filamentous fungi.
- Source :
-
Fungal genetics and biology : FG & B [Fungal Genet Biol] 1998 Jun-Jul; Vol. 24 (1-2), pp. 3-13. - Publication Year :
- 1998
-
Abstract
- The fungal cell wall, though phylogenetically variable, acts universally as a potent barrier to probing intracellular structures. Thus, the use of high-molecular-weight probes such as antibodies and lectins has proven a formidable challenge. We have devised a preparative method for use with various affinity probes that can be applied to a broad spectrum of filamentous fungal species and used for imaging whole cells. In this study, confocal imaging of whole-mount fungal hyphae after freeze substitution, methacrylate embedment/de-embedment, and infiltration with affinity probes has yielded remarkably improved renderings of the three-dimensional distribution of both microtubules (using antibodies against both alpha- and beta-tubulin) and concanavalin A binding sites. Using this protocol we have been able to document: (1) the three-dimensional distribution of microtubules in all regions of hyphae, (2) the presence of apparent foci for cytoplasmic microtubules, (3) persistent cytoplasmic microtubules during mitosis, and (4) a three-dimensional view of many compartments of the endomembrane system including Golgi-equivalent organelles and apical vesicles. The last result represents the first direct confirmation of apical vesicles comprising the Spitzenkörper.<br /> (Copyright 1998 Academic Press.)
Details
- Language :
- English
- ISSN :
- 1087-1845
- Volume :
- 24
- Issue :
- 1-2
- Database :
- MEDLINE
- Journal :
- Fungal genetics and biology : FG & B
- Publication Type :
- Academic Journal
- Accession number :
- 9742188
- Full Text :
- https://doi.org/10.1006/fgbi.1998.1054