An Improved Silver Stain for Developing Nervous Tissue

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Abstract
A reduced Ag technique using physical development to stain (chick and tree shrew [Tupaia]) embryonic nervous tissue is described. Brains are fixed in Bodian''s fixative. Paraffin sections are pretreated 100% chromic acid or 5% formol. They are impregnated with 0.01% AgNO3 dissolved in 0.1 M boric acid/sodium tetraborate buffer of pH 8 or with silver proteinate. They are developed in a special physical developer which contains 0.1% AgNO3, 0.01-0.1% formol as reducing agent, 2.5% sodium carbonate to buffer the solution at pH 10.3, 0.1% NH4NO3 to prevent precipitation of Ag(OH)2, and 5% tungstosilicic acid as a protective colloid. The development takes several minutes in this solution, thus the intensity of staining can easily be controlled. The method yields uniform, complete and reproducible staining of axons at all developmental stages of the nervous tissue and is easy to handle.