A Golgi impregnation technique for thin brain slices maintained in vitro.

A technique for using routine rapid Golgi impregnation procedures on very thin freshly fixed slices (less than 0.5 mm) of brain tissue is described. The technique was particularly successful with hippocampal slices that were maintained and stimulated in vitro prior to fixation. Thin tissue slices were surrounded by thicker sections of tissue to form a 5 mm thick bundle. The tissue bundle was then processed by a rapid Golgi procedure, 5 days each in chromate osmium and silver nitrate solutions. At the end of this time the thin tissue slices were unwrapped from their thicker protecting tissue sections, embedded in celloidin, cut at 60-100 micrometer thickness on a sliding microtome and mounted in permount under cover glass. Qualitative light microscopic analysis of the rapid Golgi impregnated slices revealed fully impregnated cell bodies, dendrites, dentritic spines, axons and axonal varicosities with minimal background artifact. In contrast, unprotected thin tissue slices showed only a dense black artifact without cells or processes.