A RAPID TECHNIQUE OF ACETYLCHOLINESTERASE STAINING

When tissue samples are evaluated for the presence of Hirschsprung's d isease, results of acetylcholinesterase staining are usually more info rmative than findings from the routine hematoxylin-eosin method. The s tandard technique for staining acetylcholinesterase takes 2 hours to p erform and is based on the enzyme histochemical method of Karnovsky an d Roots, with a subsequent visualization step as described by Hanker a nd colleagues. We modified the technique by the introduction of supple mental oxidation, which accelerated the staining time to less than 10 minutes. When evaluated in 10 patients with Hirschsprung's disease and in seven control patients with normal colons, the rapid technique gav e identical staining of nerves and ganglion cells to that of the stand ard method. Increased visualization of endogenous peroxidase-containin g histiocytes occurred, but it was not a diagnostic problem. The techn ique is sufficiently rapid to be used in conjunction with routine froz en sections to assist in the diagnosis and in selecting the optimal le vel of resection at the time of definitive surgery.