Tissue section AFM: In situ ultrastructural imaging of native biomolecules

Helen K. Graham, Nigel W. Hodson, Judith A. Hoyland, Sarah J. Millward-Sadler, David Garrod, Anthea Scothern, Christopher E M Griffiths, Rachel E B Watson, Thomas R. Cox, Janine T. Erler, Andrew W. Trafford, Michael J. Sherratt

    Research output: Contribution to journalArticlepeer-review

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

    Keywords

    • Atomic force microscopy
    • Collagen fibrils
    • Elastic fibres
    • Native biomolecules
    • Tissue cryo-sections

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