Technical pitfalls potentially affecting diagnoses in immunohistochemistry

Abstract

Result of the immunohistochemical reactions routinely used in diagnostic surgical pathology should be properly interpreted, since false results, related to technical and interpretative pitfalls may lead to incorrect diagnosis. We review of the main sources of such pitfalls, analytically described and related to different steps (fixation, tissue processing and embedding, decalcification, antigen retrieval) which may affect the accuracy of immunohistochemistry. In addition, presence of endogenous enzyme activity, improper binding of avidin to endogenous biotin, incorrect use of antibodies, chromogen and detection system, as well as incorrect interpretation may produce unreliable data. The high frequency and extension of such pitfalls make mandatory the use of internal and external controls and adoption of cross-validation programs. The present study, supported by an extensive review of the related literature is intended as a guideline leading to proper interpretation of immunohistochemical data, an essential component of the diagnostic process. Our experience on the antigen retrieval procedures for different antigens is also presented.