CAP Pro Course - Histology - Immunohistochemistry Part 1 (2025 & 2026)

Author: Carla Shoffeitt, MSM, HT(ASCP)
Reviewer: Laurie Bjerklie, MA, MLS(ASCP)CM

Continuing Education Credits

Objectives

  • Discuss the basic principles of immunohistochemistry (IHC).
  • Review fixation and processing requirements related to IHC.
  • Identify and discuss methods of epitope retrieval.
  • Discuss methods and detection systems used in the visualization of antibody-antigen reactivity.
  • Outline basic IHC staining protocol and staining considerations.
  • Discuss quality control (QC) measures used in IHC, including actions taken when QC results are not acceptable.

Course Outline

  • Discuss the basic principles of immunohistochemistry (IHC).
      • Defining Immunohistochemistry
      • Antibodies, Antigens, and Epitopes
      • Antibody-Antigen Binding
      • Monoclonal versus Polyclonal Antibodies
      • Antibody Dilution and Titration
      • Jane is preparing serial titrations of a new concentrated antibody. She has diluted the antibody with a proteinaceous, buffered solution. This solutio...
      • You have been asked to research various antibodies for C4d to be used for IHC staining of transplant heart and kidney biopsies. You find that C4d is a...
  • Review fixation and processing requirements related to IHC.
      • Fixative Considerations Based on Specimen Type
      • Formalin Fixation
      • Fixation and Processing Considerations for Breast Markers
      • Microtomy and Slide Preparation
      • Amy receives a needle core biopsy of a breast mass on Tuesday morning. Upon review of the accompanying requisition, Amy sees that the time the breast ...
      • Microtomy and slide preparation can affect the staining quality of IHC staining. Which of these considerations specifically related to slide prep is i...
      • What type of fixation method should be used for blood smears, touch-preps, and cytocentrifuged cytology preparations to ensure accurate staining?
  • Identify and discuss methods of epitope retrieval.
      • Unmasking of Epitopes
      • Proteolytic-Induced Epitope Retrieval
      • Heat-Induced Epitope Retrieval
      • An IHC-stained slide has been returned to you by the pathologist due to concern with the staining pattern on the control tissue. Upon review, you conf...
      • Drew has been asked to provide an educational presentation on immunohistochemistry. When considering the discussion on epitope retrieval, which of the...
  • Discuss methods and detection systems used in the visualization of antibody-antigen reactivity.
      • Direct and Indirect Methods of Immunohistochemical Staining
      • Peroxidase Anti-Peroxidase Method
      • Avidin-Biotin Complex and Labeled Streptavidin-Biotin Methods
      • Polymer-Based Immunohistochemistry
      • Your laboratory is evaluating the purchase of a new automated IHC staining instrument. You have learned that the vendor for the instrument is promotin...
      • Carson works in a research laboratory that will soon start immunohistochemistry (IHC) staining. Carson has been reviewing the various methods of IHC s...
      • When performing immunohistochemistry (IHC) staining with a polymer-based staining method, you observe that the staining protocol does not include a st...
  • Outline basic IHC staining protocol and staining considerations.
      • Blocking Non-Specific Binding
      • Importance of Washing Steps
      • Incubation of Primary Antibody and Detection System Reagents
      • Chromogen and Substrates
      • Maria is trying to determine which enzyme substrate-chromogen combination should be used in her IHC laboratory for staining that is non-soluble and su...
      • Trina is optimizing a new antibody that requires dilution and multiple titrations. When determining the incubation period for the various dilutions, w...
  • Discuss quality control (QC) measures used in IHC, including actions taken when QC results are not acceptable.
      • Controls in Immunohistochemistry
      • Validation of New Primary Antibodies
      • Validating New Lots of Antibody and Ancillary Reagents
      • Storage Considerations for Immunohistochemistry Reagents
      • Documenting Instrument Function and Maintenance
      • Participation in Quality Assessment Programs
      • The definition of test validation is the demonstration that a test system works in the manner in which it was intended. In immunohistochemistry (IHC),...
      • Which of the following is most important when choosing a positive control?
      • You have just received a new antibody in the laboratory. After diluting and aliquoting the antibody, how would you store the aliquots?
  • References
      • References

Additional Information

Level of Instruction: Intermediate
Intended Audience: This program is designed as an educational and training tool for Histotechnologists, Histotechnicians, Medical Laboratory Scientists, and Medical Laboratory Technicians, as well as medical laboratory science and histology students, pathology residents, and practicing pathologists.
Author Information: Carla J. Shoffeitt, MSM, HT(ASCP), is the System Director of Anatomic Pathology for Emory Healthcare in Atlanta, Georgia. She holds a Master of Science Degree in Healthcare Management as well as a Bachelor of Science degree in Biology from Troy University, Troy, Alabama. She is certified as a Histotechnician and has 29 years of experience in the field of Histology.
Reviewer Information: Laurie Bjerklie, MA, MLS(ASCP)CM, is currently an Education Developer. She earned a B.S. in Medical Laboratory Science from the University of North Dakota and an M.A. in Curriculum and Instruction from Saint Xavier University. She has over 15 years of experience in higher education and has held program director and faculty positions in both MLT and MLS programs.

This course is part of the CAP Competency Assessment Hub.

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