Tissue section AFM: In situ ultrastructural imaging of native biomolecules

Research output: Contribution to journalArticle

  • External authors:
  • Helen K. Graham
  • Nigel W. Hodson
  • Sarah J. Millward-Sadler
  • David Garrod
  • Anthea Scothern
  • Thomas R. Cox
  • Janine T. Erler

Abstract

Conventional approaches for ultrastructural high-resolution imaging of biological specimens induce profound changes in bio-molecular structures. By combining tissue cryo-sectioning with non-destructive atomic force microscopy (AFM) imaging we have developed a methodology that may be applied by the non-specialist to both preserve and visualize bio-molecular structures (in particular extracellular matrix assemblies) in situ. This tissue section AFM technique is capable of: i) resolving nm-μm scale features of intra- and extracellular structures in tissue cryo-sections; ii) imaging the same tissue region before and after experimental interventions; iii) combining ultrastructural imaging with complimentary microscopical and micromechanical methods. Here, we employ this technique to: i) visualize the macro-molecular structures of unstained and unfixed fibrillar collagens (in skin, cartilage and intervertebral disc), elastic fibres (in aorta and lung), desmosomes (in nasal epithelium) and mitochondria (in heart); ii) quantify the ultrastructural effects of sequential collagenase digestion on a single elastic fibre; iii) correlate optical (auto fluorescent) with ultrastructural (AFM) images of aortic elastic lamellae. © 2010 International Society of Matrix Biology.

Bibliographical metadata

Original languageEnglish
Pages (from-to)254-260
Number of pages6
JournalMatrix Biology
Volume29
Issue number4
DOIs
Publication statusPublished - May 2010