Comparing digital histology slides with multiple staining based on decoloring and dyeing technique

Abstract

Information in histology slides are usually visualized by different staining techniques, each of them unveils specific chemical and biological substances within tissue samples. Correlations between different stains can be useful to predict how certain tissue slides may look like if they were stained by other staining techniques. This work investigates two stains including hematoxylin and eosin (H&E) and immunohistochemistry (IHC) in digital pathological slides. Four cases of surgical biopsies were used in this work. The specimens were subjected to two consecutive stains with a decoloring process based on ethanol and potassium permanganate in between. After each stain, slides were digitized and archived as results. Comparing the effects of the two staining pipelines, IHC slides after decoloring of H&E showed that the cell structure was clear, the positive IHC staining was accurate, the background of the slide was clean, there was no DAB residue, and tissue fragments were intact. However, the other pipeline where IHC was stained before H&E showed that the nuclear border was blurred. Eosin is lightly colored resulting in low contrast visualization of nucleoplasm, DAB is not completely decolored, and parts of tissue were fragmented. We conclude that, from the proposed staining and decoloring technique, tissue slides could be stained with IHC more effectively on decolored H&E slides than those stained with H&E after IHC. Utilizing digital section scanning technology, we can obtain pairs of tissue images stained differently while preserving the exact same tissue structure.