Ideal site of biopsy in Hirschsprungs disease

The typical findings of Hirschsprung’s disease include absence of ganglion cells. What other stain can help support the diagnosis, and what is the ideal site of biopsy? 

Acetylcholinesterase stain highlights the proliferation of thickened nerve fibers in the lamina propria and muscularis mucosae. This stain is done on the frozen tissue. So, ideally, two biopsy samples are sent—one in formalin and another fresh for freezing. The site of biopsy is at least 2 cm above the dentate line. The lower rectum (adjacent to dentate line) is physiologically hypoganglionic. Also, submucosa should be included in the biopsy samples to assess nerves in both the lamina propria and muscularis mucosae.

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