Fixation and Staining of Fungus Hyphae and Host Plant Root Tissues for Electron Microscopy
1966; Taylor & Francis; Volume: 41; Issue: 1 Linguagem: Inglês
10.3109/10520296609116276
ISSN0038-9153
Autores Tópico(s)Plant Pathogens and Fungal Diseases
ResumoA deuteromycetous root-infecting fungus (Pyrenochaeta terrestris) and one of its hosts (Allium cepa) were fixed in OsO4, OsO4 vapor, KMnO4, glutaraldehyde followed by OsO4, acrolein followed by OsO4 and a glutaraldehyde-acrolein mixture followed by OsO4. The fungus was also fixed in LiMnO4. Three procedures were selected for routine comparisons because they provided suitable detail of organelles within cells of both the host and the pathogen; they were as follows: (1) 1% OsO4 for 2 hr in an ice bath; (2) 3% glutaraldehyde with 3% acrolein or 3-6% acrolein for 1-2 hr at room temperature then 1% OsO4 for 2 hr in an ice bath; and (3) 3% KMnO4 for 2 hr at room temperature. These fixatives were buffered with 0.2 M sodium cacodylate at pH 7.2-7.4. Fixation was followed by soaking the tissue blocks for a few hours to 2 days in 0.5% uranyl acetate, then by dehydration and embedding. Sections were stained with Reynolds' lead citrate. Glutaraldehyde-acrolein or acrolein provided the most detail of different organel...
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