A Comparison of Techniques Useful for Preparing Nematodes for Scanning Electron Microscopy

Abstract

Second-stage juveniles of Meloidogyne incognita were prepared by several different techniques for scanning electron microscopy (SEM). Sequential fixation in the cold (4-8 C) was superior to rapid fixation at room temperature, glutaraldehyde and glutaraldehyde-formalin were better fixatives than formalin alone, and critical point drying with carbon dioxide or Freon gave similar results that were only slightly better than air drying with Freon. Freeze drying sequentially fixed nematodes from 100% ethanol in liquid propane produced the best preserved specimens with the fewest artifacts. Specimens of various free-living and plant-parasitic nematodes were prepared for SEM by freeze drying. This technique was adequate for most genera but unsatisfactory for a few. Although each genus may require a different procedure for optimum preservation of detail, sequential fixation with glutaraldehyde and freeze drying are comparable and often superior to commonly used techniques for preparing nematodes for SEM. Key words: critical point drying, fixation, fixatives, freeze drying, methods, morphology, Acrobeles sp., Belonolaimus longicaudatus, Criconemella sp., Hemicycliophora sp., Heterodera glycines, Hoplolaimus sp., Meloidogyne incognita, M. pini, Mononchus sp.