Visualizing In Vitro Type I Collagen Fibrillogenesis by Transmission Electron Microscopy
Techniques and protocols for the in vitro formation of collagen type I fibrils and the extensive biochemical variation of the fibrillogenesis conditions are presented. In all cases, the incubation and fibrillogenesis product can be readily monitored by transmission electron microscopic study of negatively stained specimens. Representative TEM data is presented and discussed within the context of the products of the fibrillogenesis protocols, from which the extensive biochemical and structural possibilities of this integrated approach can be appreciated.
Key wordsCollagen type I Heterotrimer Fibrillogenesis Negative staining Electron microscopy D-Banded fibril Tactoid Segment long spacing (SLS) crystallite ATP Polyphosphate Sulfonated diazo dye
This and related work was performed by the author as a scientific staff member in the laboratories of the Institute of Zoology, University of Mainz, Germany, with the support of Professor Juergen Markl, and also in retirement using equipment in EM Research Services at Newcastle University, UK, with the support of Professor Richard J. Lewis, Institute of Cellular and Molecular Biosciences.
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