Dot Blotting, Electrophoresis and the three Transfers (Southern, Northern a d Western) are widely used techniques in Molecular Biology. The sensitivities of detection could be improved by many orders of magnitude if the macromolecules to be probed were confined to a small area and instead of visible staining or radiolabeling, colloidal gold labeling and electron microscopic observation were substituted.
The aim of the project is to develop modifications of these techniques to make such a gain possible. Minute Blots will be deposited from micropipets. For separations, Capillary Zone Electrophoresis will be used. Probes will be biotinylated for binding of Streptavidin coated colloidal gold; antibodies will be colloidal gold labeled. Probing will be done as conventionally; the specimens will be examined in a SEM with Back Scattered Electron detector. If these microtechniques can be successfully developed single cell Molecular Biology is expected to be feasible.
