SpringerProtocols: Abstract: ECM Macromolecules: Rotary Shadowing and Transmission Electron Microscopy

Contents of this article

Browse by Subject

Upload a Protocol

Upload your own protocols for personal use.

Protocol Alert

Receive e-mail notifications about new content on SpringerProtocols.

Video Protocols

View our video protocols.

Call for video protocols.

Comments

Read comments by other users and add your own.

Favorites

Save your favorite protocols to your My Protocols area.

RSS

RSS Feeds

Inside SpringerProtocols

Useful Tools

11. ECM Macromolecules: Rotary Shadowing and Transmission Electron Microscopy

By: Michael J. Sherratt¹

, Roger S. Meadows, Helen K. Graham, Cay M. Kielty, David F. Holmes

Abstract

Full Text

Download PDF (302K)

Conventional preparation techniques for electron microscopy employ contrast enhancing heavy metal stains in solution to visualize isolated macromolecules. In rotary shadowing electron microscopy, the heavy metal is evaporated onto surface adsorbed molecules and macromolecular assemblies. High resolution shadowing remains a valuable method for the visualization and characterization of extracellular matrix macromolecules including fibrillar collagens, microfibrillar elements, and glycoproteins.

Affiliation(s): (1) Faculty of Life Sciences, The Medical School, Stopford Building, The University of Manchester, Oxford Road, Manchester, M13 9PT, UK

Book Title: Extracellular Matrix Protocols: Second Edition

Series: Methods in Molecular Biology | Volume: 522 | Pub. Date: Jun-01-2008 | Page Range: 1-7 | DOI: 10.1007/978-1-59745-413-1_11

Subject: Cell Biology

Key Words: Rotary shadowing - Electron microscopy - Carbon film - Type I collagen - Fibrillin microfibrils - Type VI collagen microfibrils

References

Download References

1.	Myers, J.C, Amenta, P.S., Dion, A.S., Sciancalepore, J.P., Nagaswami, C and Weisel, J.W., Yurchenco, P.D.(2007)The molecular structure of human tissue type XV presents a unique conformation among the collagens. Biochem J.404, 535–44.

2.	Carvalho, H.F., Felisbino, S.L., Keene, D.R. and Vogel, K.G.(2006)Identification, content, and distribution of type VI collagen in bovine tendons. Cell Tissue Res.325, 315–24.

3.	Lethias, C., Carisey, A., Comte, J., Cluzel, C. and Exposito, J.Y.(2006)A model of tenascin-X integration within the collagenous network. FEBS Lett.580, 6281–5.

4.	Kobayashi, N., Kostka, G., Garbe, J.H., Keene, D.R., Bachinger, H.P., Hanisch, F.G., Markova, D., Tsuda, T., Timpl, R., Chu, M.L. and Sasaki, T.(2007)A comparative analysis of the fibulin protein family. Biochemical characterization, binding interactions, and tissue localization. J Biol Chem.282, 11805–16.

5.	Kim, J.K., Xu, Y., Xu, X., Keene, D.R., Gurusiddappa, S., Liang, X., Wary, K.K. and Hook, M.(2005)A novel binding site in collagen type III for integrins alpha(1)beta(1) and alpha(2)beta(1). J Biol Chem.280, 32512–20.

6.	Holden, P., Meadows, R.S., Chapman, K.L., Grant, M.E., Kadler, K.E. and Briggs, M.D.(2001)Cartilage oligomeric matrix protein interacts with type IX collagen, and disruptions to these interactions identify a pathogenetic mechanism in a bone dysplasia family. J Biol Chem.276, 6046–55.

7.	Mould, A.P., Holmes, D.F., Kadler, K.E., and Chapman, J.A.(1985)Mica sandwich technique for preparing macromolecules for rotary shadowing. J Ultrastruct Res.91, 66–76.

Comments (Loading...)

Post comment/View all comments