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Related Concept Videos

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Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Getting controls under control: the time is now for immunohistochemistry.

Emina Emilia Torlakovic1, Søren Nielsen2, Mogens Vyberg2

  • 1Department of Laboratory Hematology, Toronto General Hospital, University Health Network, Toronto, Ontario, Canada Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada Canadian Immunohistochemistry Quality Control (CIQC), Vancouver, BC, Canada.

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Summary

Diagnostic immunohistochemistry (dIHC) requires standardization. This study discusses the importance of external controls and reference standards for improving dIHC test performance and overall standardization.

Keywords:
IMMUNOHISTOCHEMISTRYIMMUNOPHENOTYPINGQUALITY CONTROLSTAINING

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Area of Science:

  • Biomedical diagnostics
  • Laboratory medicine
  • Histopathology

Background:

  • Diagnostic immunohistochemistry (dIHC) has historically been viewed as an art, lacking rigorous scientific test performance characterization.
  • Key test performance characteristics for dIHC have not been clearly defined, hindering its recognition as a standardized laboratory test.
  • The absence of standardized controls and reference standards has contributed to variability and a lack of reproducibility in dIHC.

Purpose of the Study:

  • To highlight the critical role of standardization in diagnostic immunohistochemistry (dIHC).
  • To discuss the necessity of defining and evaluating relevant test performance characteristics for dIHC.
  • To emphasize the importance of standardized external controls and reference standards for dIHC.

Main Methods:

  • Literature review and conceptual discussion on the standardization of immunohistochemistry.
  • Analysis of the impact of undefined test performance characteristics on dIHC.
  • Exploration of the role of external controls and reference standards in IHC standardization.

Main Results:

  • Diagnostic IHC has lacked clear definitions for test performance characteristics.
  • There is a significant need for standardized controls and reference standards in dIHC.
  • Standardization efforts are crucial for establishing dIHC as a reliable laboratory test.

Conclusions:

  • Standardizing external controls is essential for defining dIHC test performance characteristics.
  • Standardized controls and reference standards are fundamental for the overall standardization of IHC.
  • Implementing standardization will enhance the reliability and reproducibility of diagnostic IHC.