Flechsler, Jennifer and Heimerl, Thomas and Pickl, Carolin and Rachel, Reinhard and Stierhof, York-Dieter and Klingl, Andreas (2020) 2D and 3D immunogold localization on (epoxy) ultrathin sections with and without osmium tetroxide. MICROSCOPY RESEARCH AND TECHNIQUE, 83 (6). pp. 691-705. ISSN 1059-910X, 1097-0029
Full text not available from this repository. (Request a copy)Abstract
For nearly 50 years immunogold labeling on ultrathin sections has been successfully used for protein localization in laboratories worldwide. In theory and in practice, this method has undergone continual improvement over time. In this study, we carefully analyzed circulating protocols for postembedding labeling to find out if they are still valid under modern laboratory conditions, and in addition, we tested unconventional protocols. For this, we investigated immunolabeling of Epon-embedded cells, immunolabeling of cells treated with osmium, and the binding behavior of differently sized gold particles. Here we show that (in contrast to widespread belief) immunolabeling of Epon-embedded cells and of cells treated with osmium tetroxide is actually working. Furthermore, we established a "speed protocol" for immunolabeling by reducing antibody incubation times. Finally, we present our results on three-dimensional immunogold labeling.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ELECTRON-MICROSCOPY; IGNICOCCUS-HOSPITALIS; COLLOIDAL GOLD; PHAEODACTYLUM-TRICORNUTUM; ULTRASTRUCTURAL ANALYSIS; TANNIC-ACID; PROTEIN; MEMBRANE; LIGHT; PRESERVATION; (serial) immunogold labeling; Archaea; Diatoms; Epon 812-osmium tetroxide |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) > Prof. Dr. Reinhard Rachel |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 30 Mar 2021 08:30 |
| Last Modified: | 30 Mar 2021 08:30 |
| URI: | https://pred.uni-regensburg.de/id/eprint/45112 |
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