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Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
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Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

Published on: April 8, 2016

An overview of the complexities and subtleties of immunohistochemistry

A J Mighell1, W J Hume, P A Robinson

  • 1Leeds Dental Institute, UK. A.J.Mighell@leeds.ac.uk

Oral Diseases
|February 11, 1999
PubMed

Insights

This review highlights critical considerations for immunohistochemistry (IHC) techniques. Proper planning, antibody selection, and control interpretation are essential for reliable research findings using IHC.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Histology

Background:

  • Immunohistochemistry (IHC) is a valuable research method.
  • IHC studies often overlook critical technical complexities.
  • Inconsistent application limits the reliability of IHC results.

Purpose of the Study:

  • To identify common problems and limitations in immunohistochemistry.
  • To outline essential procedures for planning and interpreting IHC studies.
  • To emphasize the importance of technique standardization in IHC.

Main Methods:

  • Review of literature on immunohistochemistry best practices.
  • Discussion of factors influencing IHC staining.
  • Analysis of critical control requirements for IHC.

Main Results:

  • Alternative splicing and post-translational modifications generate protein isoforms.
  • Primary antibody selection significantly impacts IHC outcomes.
  • Tissue processing (fixation, antigen retrieval) and controls are crucial for accurate interpretation.

Conclusions:

  • Standardized protocols are necessary for robust immunohistochemistry.
  • Careful consideration of technical variables enhances IHC reliability.
  • Proper interpretation of IHC staining patterns is vital for valid conclusions.

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