Direct visualization and silver enhancement of ultra-small antibody-bound gold particles on immunolabeled ultrathin resin sections

Details

Serval ID
serval:BIB_07A72E8C9256
Type
Article: article from journal or magazin.
Collection
Publications
Title
Direct visualization and silver enhancement of ultra-small antibody-bound gold particles on immunolabeled ultrathin resin sections
Journal
Scanning Microscopy
Author(s)
Stierhof Y.D., Humbel B.M., Hermann R., Otten M.T., Schwarz H.
ISSN
0891-7035
Publication state
Published
Issued date
1992
Volume
6
Number
4
Pages
1009-1022
Language
english
Abstract
Ultra-small gold colloids bound to immunolabeled ultrathin resin sections were visualized using transmission, scanning, and scanning transmission electron microscopy (TEM, SEM, STEM). The best marker contrast is obtained in a field emission STEM (200 kV) equipped with a high-angle annular dark-field (HAADF) detector. HAADF STEM renders possible the simultaneous visualization of ultra-small gold and ultrastructural details in unstained resin sections, and an overall presentation of a labeled E. coli cell.
For routine work, an enhancement step is a pre-requisite for easy detection of bound marker molecules. Five different silver enhancing solutions were tested for their suitability for ultra-small gold intensification. Enhancers lacking the protective colloid gum arabic exhibit lower quality with regard to efficiency and homogeneity of enhancement. This problem can be overcome by adding gum arabic. Silver enhancement generally results in heterogeneously sized particles. This is most probably due to the heterogeneous original gold colloid probe. In general, an estimation of enhancement efficiency is associated with difficulties depending on experimental conditions and the electron microscopic imaging modes used. Only a low number of the ultra-small gold particles seems to remain unenhanced or poorly enhanced when treated with ''high-quality'' enhancers. On-section labeling of ultrathin resin sections with silver-enhanced ultra-small gold markers also offers the possibility of high-resolution immunolabeling experiments at the light microscopic level.
Keywords
1 NM GOLD, SILVER ENHANCEMENT EFFICIENCY, TRANSMISSION ELECTRON MICROSCOPY, FIELD EMISSION SCANNING ELECTRON MICROSCOPY, FIELD EMISSION SCANNING TRANSMISSION ELECTRON MICROSCOPY, HIGH-ANGLE ANNULAR DARK-FIELD IMAGING, IMMUNOLABELING, AUTOMETALLOGRAPHY, IMMUNOELECTRON MICROSCOPY, BACKSCATTERED ELECTRON IMAGING
Web of science
Create date
18/10/2012 13:34
Last modification date
20/08/2019 12:30
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