Imaging internal features of whole, unfixed bacteria

Nicholas M. Thomson, Kevin Channon, Noor Azlin Mokhtar, Lech Staniewicz, Ranjana Rai, Ipsita Roy, Shun Sato, Takeharu Tsuge, Athene M. Donald, David Summers, Easan Sivaniah*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Wet scanning-transmission electron microscopy (STEM) is a technique that allows high-resolution transmission imaging of biological samples in a hydrated state, with minimal sample preparation. However, it has barely been used for the study of bacterial cells. In this study, we present an analysis of the advantages and disadvantages of wet STEM compared with standard transmission electron microscopy (TEM). To investigate the potential applications of wet STEM, we studied the growth of polyhydroxyalkanoate and triacylglycerol carbon storage inclusions. These were easily visible inside cells, even in the early stages of accumulation. Although TEM produces higher resolution images, wet STEM is useful when preservation of the sample is important or when studying the relative sizes of different features, since samples do not need to be sectioned. Furthermore, under carefully selected conditions, it may be possible to maintain cell viability, enabling new types of experiments to be carried out. To our knowledge, internal features of bacterial cells have not been imaged previously by this technique.

Original languageEnglish (US)
Pages (from-to)59-68
Number of pages10
JournalScanning
Volume33
Issue number2
DOIs
StatePublished - Mar 1 2011

Keywords

  • hydrated
  • microscopy
  • polyhydroxyalkanoate
  • wet STEM
  • whole cells

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Instrumentation

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