Silver Staining of Insect Central Nervous Systems by the Bodian Protargol Method

Abstract

Improved fixation of ganglia of the central nervous system of Periplaneta americana and Schistocerca gregaria for silver staining by Power's (1943) modification of the Bodian protargol method is given by alcoholic Bouin aged for at least 40 days at 60° C. During impregnation of sections, increased copper and decreased pH give paler staining, more selective for nerve fibres. Prolonging impregnation from 24 to 48 hours weakens the stain and decreases selectivity. The intensity of the stain depends chiefly upon the amount of unreduced (developable) silver combined with the tissues; selectivity is determined mainly by the number and distribution of the reduced silver particles (‘nuclei’). In development, increased sodium sulphite gives more differentiation, increased hydroquinone gives less. Optimum developer composition depends upon impregnation, and thick sections need more differentiation than thinner ones. Within limits, change in one of the factors that control staining can be balanced by changes in others, but by suitable adjustment of the conditions the result can be varied from almost total staining of nerve fibres, for general neuroanatomy, to highly selective staining for tracing individual fibres.

The Antennal Lobe of Orthoptera – Anatomy and Evolution

“…Reduced Silver Impregnation For survey histology dissected brains were fixed either in alcoholic Bouin's [Gregory, 1970] or in acetic acid-alcohol-formaldehyde (AAF) solution [Lillie, 1965]. Brains were embedded in paraffin and sectioned at 10 µm.…”

The clypeal glands of Mynoglenes and of some other linyphiid spiders

“…Anaesthetized spiders were dissected rapidly into Gregory's Dubosq-Brasil solution (Gregory, 1970) and fixed for 24 hours. Wax sections were stained with Massons's Trichrome method, with various reduced silver methods (Blest, 1961(Blest, , 1976a for ductules, and with a number of routine histological techniques.…”

Cytologic and immunocytochemical findings of anaplastic large cell lymphoma

“…The optic lobes of over 40 animals were stained by a variety of selective silver techniques (Rowell's procedure [Rowell, 1963], Holmes‐Blest [Blest, 1961], Bodian Protargol [Gregory, 1970], and Nonidez method [Nonidez, 1939]) as well as standard toluidine blue. The most reliable and effective methods tried were Rowell's procedure and the Nonidez method.…”