Cationic hydrous thorium dioxide colloids--a useful tool for staining negatively charged surface matrices of bacteria for use in energy-filtered transmission electron microscopy.
BACKGROUND
Synthesis of cationic hydrous thorium dioxide colloids (ca. 1.0 to 1.7 nm) has been originally described by Müller 22 and Groot 11 and these have been used by Groot to stain acidic glucosaminoglycans for ultrastructure research of different tissues by conventional transmission electron microscopy.


RESULTS
Synthesis of colloidal thorium dioxide has been modified and its use as a suitable stain of acidic mucopolysaccharides and other anionic biopolymers from bacteria, either as whole mount preparations or as preembedment labels, is described. The differences in stain behavior relative to commonly used rutheniumred-lysine and Alcian Blue electron dense acidic stains has been investigated and its use is exemplified for Pseudomonas aeruginosa adjacent cell wall biopolymers. For the first time thorificated biopolymers, i.e. bacterial outer cell wall layers, have been analysed at the ultrastructural level with electron energy loss spectroscopy (EELS) and electron spectroscopic imaging (ESI), leading to excellent contrast and signal strength for these extracellular biopolymers.


CONCLUSIONS
Application of cationic hydrous ThO2 colloids for tracing acidic groups of the bacterial surface and/or EPS has been shown to be rather effective by transmission electron microscopy. Because of its high electron density and its good diffusibility it stains and outlines electro-negative charges within these biopolymers. In combination with ESI, based on integrated energy-filtered electron microscopy (EFTEM) Th-densities and thus negative charge densities can be discriminated from other elemental densities, especially in environmental samples, such as biofilms.
Lünsdorf
Heinrich
H
GBF-National Institute of Biotechnology, Department of Environmental Microbiology, Mascheroder Weg 1, D-38124 Braunschweig, Germany. hlu@gbf.de
Kristen
Ingeborg
I
Barth
Elke
E
D000818:Animals; D000942:Antigens, Bacterial / Q000378:metabolism; D015373:Bacterial Typing Techniques; D046032:Microscopy, Energy-Filtering Transmission Electron; D013194:Staining and Labeling; D013911:Thorium Dioxide
D000942:Antigens, Bacterial; D013911:Thorium Dioxide