Chapter 7 Dense Tissue and Special Stains

Abstract

Publisher Summary This chapter presents data concerning the selective staining method combined with high-voltage electron microscopy to analyze the three-dimensional structure within dense tissue. The Golgi impregnation technique is an excellent method of revealing the three-dimensional morphology of neurons and glia cells and it has been used extensively by light microscopists to explore the organization of the central nervous system. In contrast to light microscopy, electron microscopy of Golgi impregnated tissue has the advantage of revealing both impregnated and impregnated cells at the same time in greater detail. It is demonstrated that the whole Golgi apparatus could be stained by this double impregnation technique, using uranyl acetate followed by lead and copper. The principle of selective staining for the T-system is to fill or stain the extracellular space with molecular tracers. Selective stains or tracers successfully used are horseradish peroxidase, lanthanum, ruthenium red, and diaminobenzidine. It is found that selective staining with 3,3′-diaminobenzidine tetrahydrochloride–ferrocyanide for smooth endoplasmic reticulum was used to visualize its distribution within 0.5- to 1-pm-thick sections of mouse and frog peripheral nerves in high-voltage electron microscopy.