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Negative staining and transmission electron microscopy of bacterial surface structures

Mörgelin, Matthias LU (2017) In Methods in Molecular Biology 1535. p.211-217
Abstract

Negative staining is an essential and versatile staining technique in transmission electron microscopy that can be employed for visualizing bacterial cell morphology, size, and surface architecture at high resolution. Bacteria are usually transferred by passive electrostatic adsorption from suspensions in physiological saline onto suitable hydrophilic support films on electron microscopic grids. There they are contrasted, or “stained,” by heavy metal ions in solution such as tungsten, uranyl, molybdate, or vanadate compounds. Here, I describe how to visualize the interaction between the bacterial M1 protein and complement factors C1q and C3 on the surface of group A streptococcus by negative staining with uranyl formate on carbon... (More)

Negative staining is an essential and versatile staining technique in transmission electron microscopy that can be employed for visualizing bacterial cell morphology, size, and surface architecture at high resolution. Bacteria are usually transferred by passive electrostatic adsorption from suspensions in physiological saline onto suitable hydrophilic support films on electron microscopic grids. There they are contrasted, or “stained,” by heavy metal ions in solution such as tungsten, uranyl, molybdate, or vanadate compounds. Here, I describe how to visualize the interaction between the bacterial M1 protein and complement factors C1q and C3 on the surface of group A streptococcus by negative staining with uranyl formate on carbon support films. The methodology should be generally applicable to the study of a large number of other bacteria-protein interactions.

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author
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
Adsorption, Bacteria, Carbon film, Contrast, Immunostaining, Negative staining, Protein interactions, Transmission electron microscopy
in
Methods in Molecular Biology
volume
1535
pages
7 pages
publisher
Humana Press
external identifiers
  • scopus:85005996169
ISSN
10643745
DOI
10.1007/978-1-4939-6673-8_13
language
English
LU publication?
no
id
89b467f1-b3a3-4ff5-b989-b3b833153db3
date added to LUP
2017-03-16 13:53:46
date last changed
2018-01-07 11:56:01
@inbook{89b467f1-b3a3-4ff5-b989-b3b833153db3,
  abstract     = {<p>Negative staining is an essential and versatile staining technique in transmission electron microscopy that can be employed for visualizing bacterial cell morphology, size, and surface architecture at high resolution. Bacteria are usually transferred by passive electrostatic adsorption from suspensions in physiological saline onto suitable hydrophilic support films on electron microscopic grids. There they are contrasted, or “stained,” by heavy metal ions in solution such as tungsten, uranyl, molybdate, or vanadate compounds. Here, I describe how to visualize the interaction between the bacterial M1 protein and complement factors C1q and C3 on the surface of group A streptococcus by negative staining with uranyl formate on carbon support films. The methodology should be generally applicable to the study of a large number of other bacteria-protein interactions.</p>},
  author       = {Mörgelin, Matthias},
  issn         = {10643745},
  keyword      = {Adsorption,Bacteria,Carbon film,Contrast,Immunostaining,Negative staining,Protein interactions,Transmission electron microscopy},
  language     = {eng},
  pages        = {211--217},
  publisher    = {Humana Press},
  series       = {Methods in Molecular Biology},
  title        = {Negative staining and transmission electron microscopy of bacterial surface structures},
  url          = {http://dx.doi.org/10.1007/978-1-4939-6673-8_13},
  volume       = {1535},
  year         = {2017},
}