Extracellular matrix reorganization during cryo preparation for scanning electron microscope imaging of staphylococcus aureus biofilms

Yong Wu, Jing Liang, Kim Rensing, Tseng Ming Chou, Matthew Libera

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

Biofilms are three-dimensional communities of bacteria distributed in a highly hydrated extracellular matrix (ECM). They can be visualized by scanning electron microscopy (SEM), but the requisite SEM sample preparation can modify the biofilm morphology. Here, four different approaches to prepare biofilms of hydrated Staphylococcus aureus for SEM imaging are compared. In order of increasing cooling effectiveness these are: (1) drying in air; (2) plunging in liquid nitrogen; (3) plunging in liquid ethane; and (4) high pressure freezing with liquid nitrogen. These different methods give rise to markedly different biofilm morphologies, which are revealed by cryo-SEM imaging. Significantly, high-pressure frozen biofilms exhibit a rich network of nanoscale ECM fibers surrounding individual bacteria throughout the biofilm thickness. This structure is entirely lost when similar biofilms are dried in air, and it is substantially modified when these biofilms are plunged into liquid nitrogen or liquid ethane.

Original languageEnglish
Pages (from-to)1348-1355
Number of pages8
JournalMicroscopy and Microanalysis
Volume20
Issue number5
DOIs
StatePublished - 11 Apr 2014

Keywords

  • S. aureus
  • biofilm
  • cryo
  • extracellular matrix
  • scanning electron microscopy

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