Preparation of prokaryotic and eukaryotic organisms using chemical drying for morphological analysis in scanning electron microscopy (SEM)

Madison A. Koon, Khadijah Almohammed Ali, Robert M. Speaker, James P. McGrath, Eric W. Linton, Michelle L. Steinhilb

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Scanning electron microscopy (SEM) is a widely available technique that has been applied to study biological specimens ranging from individual proteins to cells, tissues, organelles, and even whole organisms. This protocol focuses on two chemical drying methods, hexamethyldisilazane (HMDS) and t-butyl alcohol (TBA), and their application to imaging of both prokaryotic and eukaryotic organisms using SEM. In this article, we describe how to fix, wash, dehydrate, dry, mount, sputter coat, and image three types of organisms: cyanobacteria (Toxifilum mysidocida, Golenkina sp., and an unknown sp.), two euglenoids from the genus Monomorphina (M. aenigmatica and M. pseudopyrum), and the fruit fly (Drosophila melanogaster). The purpose of this protocol is to describe a fast, inexpensive, and simple method to obtain detailed information about the structure, size, and surface characteristics of specimens that can be broadly applied to a large range of organisms for morphological assessment. Successful completion of this protocol will allow others to use SEM to visualize samples by applying these techniques to their system.

Original languageEnglish
Article numbere58761
JournalJournal of Visualized Experiments
Volume2019
Issue number143
DOIs
StatePublished - Jan 2019

Keywords

  • Biology
  • Critical point drying (CPD)
  • Cyanobacteria
  • Drosophila
  • Euglenoid
  • Fruit fly
  • Hexamethyldisilazane (HMDS)
  • Issue 143
  • Scanning electron microscopy (SEM)
  • T-butyl alcohol (TBA)

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