HOW TO IMPROVE HISTOPATHOLOGICAL RESULTS IN THE BIOPSY DIAGNOSIS OF GUT DYSGANGLIONOSIS - A METHODOLOGICAL REVIEW

In a methodological survey, the technical prerequisites for optimal hi stopathological diagnosis of gut dysganglionosis are presented. To mak e a proper diagnosis, the pediatric surgeon or gastroenterologist and the pathologist must consider certain preconditions. The most importan t steps for the optimal biopsy diagnosis of an aganglionosis, an ultra short Hirschsprung segment, a intestinal neuronal dysganglionosis (IND ), a ganglioneuromatosis, a hypogenesis, or immaturity of the vegetati ve gut innervation are: (1) taking 3-4 biopsies the size of a pepperco rn (3-5 mm(3)) with submucosa; (2) the best instruments for taking rec tal mucosal biopsies are forceps and scissors or a conventional large biopsy forceps; and (3) biopsies may be taken 1 cm, 3-4 cm, 6-9 cm, an d 9-12 cm (or from a preternatural anus) above the pectinate line. A b iopsy containing mucosa, muscularis mucosae, and submucosa guarantees a satisfying histopathological diagnosis. The native biopsies can be t ransported on water-ice if the distance to the pathologist takes no lo nger than 4-6 h. For long distances, biopsies have to be frozen on dry ice (CO2-80 degrees C) and transported in a sufficient amount of dry ice (adapted to the time of transportation). For biopsy processing, th e following points are important: a total of 122 to 160 15-mu m-thick native cryostat serial sections have to be cut per biopsy and distribu ted on four microscope slides. Forty sections are used for lactic dehy drogenase reactions, 32 for succinic dehydrogenase reactions, and the rest for an acetylcholinesterase (AChE) reaction. An AChE reaction alo ne is sufficient for the diagnosis of Hirschsprung's disease (HD), but never for IND or other developmental malformations of the submucous a nd myenteric plexuses. Enzymehistotopochemical reactions allow the ass essment of functional parameters. These reactions, in contrast to immu nohistochemical staining, offer information about the functional activ ity of special gut structures, e. g., increased AChE activity in nerve fibers of the rectal wall in HD or a lack of dehydrogenase activity i n immature ganglia.