The diagnostic potential of autofluorescence (AF) microscopy under ultraviolet (UV) excitation is explored using ex vivo human specimens. The aim is to establish optical patterns (the rules for interpretation) that correspond to normal and abnormal histologies of the esophagus, spanning from early benign modifications (Barrett's esophagus) to subsequent dysplastic change and progression toward carcinoma. This was achieved by developing an image library categorized by disease progression. We considered morphological changes of disease as they are compared with histological diagnosis of the pathological specimen, as well as control samples of normal esophagus, proximal stomach, and small intestine tissue. Our experimental results indicate that UV AF microscopy could provide real-time histological information for visualizing changes in tissue microstructure that are currently undetectable using conventional endoscopic methods.