Quantitative ultrastructural anatomy of esophagus in different regions in the horse: effects of alternate methods of tissue processing.

A qualitative and quantitative study was made of 2 adult horses to determine the effect of specimen preparation and the influence of anatomically separate regions on the microscopic structure of the esophagus. The effects of 3 fixative solutions (formalin, Bouin's fixative and Karnovsky's fixative) and 4 histologic staining procedures (hematoxylin and eosin, Gomori's trichrome, Laidlow's reticulum, and orcein-elastin) were compared. Density-dependent image analysis was used to compare the relative contributions of elastic and reticular fibers in esophageal connective tissue for each region. Cross-sectional dimensions of the tissue layers which comprise the esophageal wall were obtained by light microscopic measurement and were compared for each region. The mucosal surface structure was examined by scanning electron microscopy. Our results indicate that specimen preparation has significant effects on the structural integrity of esophageal tissues. Bouin's fixative was inferior to the other fixatives in this regard. Furthermore, there are extensive regional differences in esophageal microstructure. Formalin-fixed tissues stained with either hematoxylin and eosin or Gomori's trichrome appear the most suitable for routine quantitative morphologic studies of the equine esophagus.