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Antibodies in Pathology: The Role of Multiplex Immunohistochemistry
Jaleh Mansouri, M.D.
While advanced molecular genetic techniques have acquired a dominant role in diagnostic
pathology, immunohistochemical (IHC) methods have remained an essential component in the
morphologic assessment of diseased tissue. In the last decade, advances in both antibody and digital
technology have optimized and expanded the applications of immunohistochemistry, especially in research
and oncologic pathology. One of these techniques, known as multiplex immunohistochemistry, involves
the use of two or more antibodies with distinct epitopes (and spectral properties) within the same tissue
section, and even within the same cell. Multiplex immunohistochemistry enables pathologists to directly
assess the differential expression of antibodies like cytokeratins within neighboring cells, and thus
increases the diagnostic accuracy of subtle lesions including atypical ductal hyperplasias or DCIS and
prostatic intraepithelial neoplasia (PIN). In the research context, multiplex immunohistochemistry allows
for colocalization of biomarkers in cellular organelles and can provide valuable insight into many disease
The main challenges in optimizing the use of these advanced IHC techniques are standardization
and validation of reagents and assays. In March of 2014, the College of American Pathologists (CAP)
issued a series of recommendations / guidelines that were published online on the Archives of Pathology and
Laboratory Medicine website addressing the need to validate antibodies and IHC assays prior to use in
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Past / Future Feature Pages:
May 2014: Computer Software and Systems
June 2014: Diagnostic Testing / Reagents
July 2014: Digital / Imaging / Photography
August 2014: Books & Journals
September 2014: Antibodies / Biomarkers
October 2014: Laboratories
November 2014: Consumable Lab Products / Clinical Lab Analyzers
December 2014: Grossing Equipment / Workstations
January 2015: Billing / Consulting / Management
February 2015: Microscopes / Microscopy Products
March 2015: Tissue Processors / Other Histology
April 2015: Laboratory Accreditation / Education / CME
May 2015: Computer Software and Systems
June 2015: Diagnostic Testing / Reagents
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