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A Dermatopathologist's Guide to Troubleshooting Immunohistochemistry Part 1: Methods and Pitfalls. | LitMetric

A Dermatopathologist's Guide to Troubleshooting Immunohistochemistry Part 1: Methods and Pitfalls.

Am J Dermatopathol

*Dermatology Resident, Department of Dermatology, University of Illinois at Chicago, Chicago, IL; and †Laboratory Manager (J.N.T.), IHC HistoTechnician (M.M.M.), Dermatopathologist (J.M.J.-H.), Director (D.M.E.), Ackerman Academy of Dermatopathology, University of Illinois, Chicago, IL.

Published: August 2015

AI Article Synopsis

  • Immunohistochemistry (IHC) is a technique used to identify specific antigens in tissue samples, using antibodies and enzymatic reactions to produce visible results.
  • The process can involve unmasking hidden epitopes that may be obscured due to formalin fixation by using heat or proteolytic enzymes.
  • The review is structured into two parts: the first discusses methodologies and challenges related to IHC testing, while the second addresses common staining issues and their solutions.

Article Abstract

Immunohistochemistry (IHC) is a method by which specific target antigens can be detected in formalin-fixed paraffin-embedded tissue and involves the use of monoclonal or polyclonal antibodies; visualization of specific tissue antigens is achieved through an enzymatic reaction that transforms a colorless chromogen to a colored one. These enzymes may be attached to the antibody through a protein-ligand method (eg, biotin-avidin or biotin-streptavidin) or through a secondary antibody. Epitopes that are masked by protein linkage during formalin fixation are unmasked using a retrieval system that either uses heat (heat-induced epitope retrieval) or proteolytic enzymes (proteolytic-induced epitope retrieval). Part 1 of this review will focus and elaborate on the available methodologies for IHC testing, common problems inherent to each technique, and how they can be resolved. Part 2 will focus on common problems and artifacts encountered during IHC staining, likely causes, and methods for addressing each problem.

Download full-text PDF

Source
http://dx.doi.org/10.1097/DAD.0000000000000335DOI Listing

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