Whole-slide imaging (WSI) contributes to medical education, collaboration, quality assurance, examination, and consultation in pathology. The images obtained from WSI are of high quality and could be stored indefinitely. In research involving esophageal squamous cell carcinoma, the combination of WSI and image processing program allows effective interpretations of expressions of various immunomarkers related to pathogenesis, prognosis, and response to therapy in tissue microarray sections. The operation and basic principles of whole-slide imaging of esophageal squamous cell carcinoma are also presented. Common use of WSI will occur with modifications of the whole-slide imaging scanners to adapt to the workflows in diagnostic and research laboratories.