title: A Dermatopathologist's Guide to Troubleshooting Immunohistochemistry Part 1: Methods and Pitfalls.
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.
authors.affiliation: *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.
authors.forename: Amanda F
authors.initials: AF
authors.lastname: Marsch
authors.forename: Jonathan N
authors.initials: JN
authors.lastname: Truong
authors.forename: Melissa M
authors.initials: MM
authors.lastname: McPherson
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Junkins-Hopkins
authors.forename: Dirk M
authors.initials: DM
authors.lastname: Elston
mesh_terms: D000906:Antibodies; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D013194:Staining and Labeling; D016707:Tissue Fixation / Q000379:methods*
chemical_list: D000906:Antibodies