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Matrix Biol. 2010 May;29(4):254-60. doi: 10.1016/j.matbio.2010.01.008. Epub 2010 Feb 6.

Tissue section AFM: In situ ultrastructural imaging of native biomolecules.

Author information

1
Unit of Cardiac Physiology, School of Biomedicine, The University of Manchester, Manchester Academic Health Science Centre, Manchester, M13 9PT, UK.

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-microm 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.

PMID:
20144712
PMCID:
PMC2877882
DOI:
10.1016/j.matbio.2010.01.008
[Indexed for MEDLINE]
Free PMC Article

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