Specific, selective, and complete staining of neurons of the myenteric plexus, using cuprolinic blue

Wholemounts consisting of both muscular layers of the rat's intestine have been stained with Cuprolinic blue and magnesium chloride. The procedure, which is specific for RNA, gives selective coloration of the somata of the neurons of the myenteric plexus. The neuronal nucleoli and Nissl substance are the only blue structures in such preparations. For descriptive and quantitative work, Cuprolinic blue is more selective than other cationic dyes, and it colors more neurons than can be shown by staining for mitochondrial NADH diaphorase. The absence of background color enables neurons to be identified more confidently and counted more quickly than is possible with other techniques, in which nuclei or mitochondria in smooth muscle and neuroglial cells are also stained. Numbers of myenteric neurons determined using the new method are higher than those obtained by other means.