Electron density imaging of protein films on gold-particle surfaces with transmission electron microscopy.
BACKGROUND
Surface bound proteins on colloid particles are widely used in biotechnological applications such as diagnostics or separation. Analysis of colloid surfaces by imaging methods provides information on the structure of these protein films, and an understanding of the functional relationships of biomolecules immobilised on solid surfaces.


METHODS
In order to visualise protein molecules organised in films on surfaces of nano-sized gold-particles, an electron-microscopic approach based on the scattering absorption contrast of the specimen was applied.


RESULTS
Analysing protein conjugated gold particles with a transmission electron microscope, protein films on gold particle surfaces cause a significant scattering absorption contrast based on the materials' electron density. Thus, the thickness of such films becomes directly measurable in planar projection and the shape of these films are visualised without negative staining methods. The insertion of Ruthenium-labelled antibodies instead of non-labelled antibodies as a marker with increased electron-density in these films yields a contrast enhancement of the whole film. Additional labelling with anti-Mouse IgG Gold conjugates localises the position of the surface bound antibodies in such protein films.


CONCLUSIONS
The power of transmission electron microscopy to resolve protein-films on colloid surfaces without staining or labelling as a sample preparation procedure has been demonstrated. Thus, this direct method provides an analytical tool for studying protein films and their structural features on particle surfaces.
Jürgens
L
L
Roche Diagnostics GmbH, Mannheim, Germany. LUDGER.JUERGENS@roche.com
Nichtl
A
A
Werner
U
U
D000818:Animals; D000906:Antibodies; D002417:Cattle; D003102:Colloids; D006046:Gold; D007089:Image Enhancement / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods*; D011506:Proteins / Q000032:analysis*; D012428:Ruthenium; D012710:Serum Albumin, Bovine / Q000032:analysis
D000906:Antibodies; D003102:Colloids; D011506:Proteins; D012710:Serum Albumin, Bovine; D006046:Gold; D012428:Ruthenium