Virtual Microscopy in Prostate Histopathology: Simultaneous Viewing of Biopsies Stained Sequentially With Hematoxylin and Eosin, and α-Methylacyl-Coenzyme A Racemase/p63 Immunohistochemistry

Histopathological diagnosis of small focus carcinomas in prostatic needle biopsies is often assisted by IHC. To make a definitive diagnosis the pathologist must compare IHC findings with hematoxylin and eosin stained tissue morphology. We introduce what is to our knowledge a new application of virtual microscopy, in which hematoxylin and eosin, and IHC stains done sequentially on the same microscope slide can be simultaneously displayed and compared on a computer screen.A total of 30 hematoxylin and eosin stained prostatic needle biopsies were scanned with a computer controlled microscope. The slides were destained and then immunostained with a cocktail of AMACR and p63 antibodies, which labels the nuclei of nonmalignant basal cells (p63) and the cytoplasm of neoplastic glandular cells suspicious for malignancy (AMACR). The slides were then scanned again and the pairs of virtual slides were aligned for synchronized viewing.The presented technique was found helpful when suspicious lesions were small and when examining the immunoprofile of specimens was warranted, in addition to examining hematoxylin and eosin stained tissue morphology. The usefulness of our approach based on virtual microscopy can be evaluated on the website , which also serves as an educational tool for self-learning the correlation between hematoxylin and eosin stained tissue morphology, and AMACR/p63 IHC in prostate biopsies.The technology for simultaneously viewing sequentially hematoxylin and eosin and IHC stained prostate biopsies can be readily used for educational purposes, as exemplified by our website, and along with the availability of rapid virtual slide scanners it can also be used for clinical diagnostics.