The application of cobalt labelling to electron microscopic investigations of serial sections

The cobalt labelling technique can be applied to ultrathin serial sections and subsequent electron microscopical investigations with the following modifications: a prolonged, up to 12 h, fixation of the tissue in aldehydes; a shortened, 15 min, postfixation in OsO4; embedding in soft resin block by using a higher proportion of plasticizer in the polimerizing mixture; mounting of 5 micrometers thick serial sections between two layers of Agar-Agar coatings; performing the intensification of the Agar section-Agar sandwich with a physical developer containing a low percentage of the reductive agent; reembedding selected thick sections for ultrathin serial sectioning and staining with uranile acetate and lead citrate. The technique unambiguously shows all labelled profiles, and preserves the fine structural details of the surrounding tissues.