A Non-toxic Scanning Electron Microscopy Method to Study the Reproductive Structures of Agaricales Using Sample Fixation in the Field
López, Giovanni J.
Velázquez, Carlos Ríos
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We present a rapid and non-toxic method to prepare lamellae and related structures from Agaricales for Scanning Electron Microscopy(SEM). The morphologies of the basidia and basidiospores of Gymnopus subpruinosus, basidiospores of Rusulla sp., and Chlorophyllum molibdites were studied in SEM to compare the efficiency of the method. The procedure consisted of an initial fixation with paraformaldehyde-gluteraldehyde (2.5%:2.0%, pH 7.2) of the specimen in the field, followed by a series of dehydrations in ethanol in the laboratory. The dehydrated samples were critical point dried, coated with gold and observed with a JEOL Scanning Electron Microscope. This process demonstrated that mycological samples such as lamellae of Agaricales (Basidiomycetes) can be prepared for SEM without rehydrating in KOH and without the use of OsO4. Micrographs of high resolution and quality were obtained with this method.