Immunohistochemistry (IHC) Basics in Histology (by LabCE)

(based on 184 customer ratings)

Authors: Diane L. Sterchi, MS, HTL(ASCP) and Donna M. Hedger, BS, HT(ASCP)
Reviewer: Tanisha Neely, HT(ASCP)

Course provided by LabCE.

This course introduces the various techniques that are used in the preparation and evaluation of immunohistochemistry (IHC) slides. Procedures and terminology related to IHC are also discussed and strategies for troubleshooting problems are presented.

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Continuing Education Credits

Florida Board of Clinical Laboratory Personnel Credit Hours - Immunohistochemistry/Advanced Histologic Techniques: 2 hour(s)
Course number 20-547723, approved through 9/1/2018


  • Explain various immunohistochemical (IHC) techniques used in the laboratory.
  • Identify appropriate quality control (QC) used while performing IHC procedures.
  • Describe slide preparation procedures used in IHC procedures.
  • Define epitope unmasking and retrieval procedures that may be used in performing IHC procedures.
  • Identify and solve problems that can occur when performing IHC procedures.

Customer Ratings

(based on 184 customer ratings)

Course Outline

  • The Role of Immunohistochemistry (IHC) in the Histology Laboratory
      • Immunohistochemistry (IHC) Uses and Considerations
      • Antibodies, Antigens, and Epitopes
  • Antibodies
      • Monoclonal Antibodies
      • Polyclonal Antibodies
      • Antibody Titering
  • Fixation
      • Tissue Handling
      • Aldehyde Fixation
      • Formalin/Aldehyde Substitutes
      • Zinc Fixatives
      • Alternative Fixatives for IHC
  • Processing and Microtomy
      • Processing Principles
      • Frozen Sections
  • Epitope Retrieval
      • General Epitope/Antigen Unmasking
      • Why Perform Epitope/Antigen Unmasking?
  • Unmasking Methods
      • Heat-Induced Epitope Retrieval (HIER)
      • Enzyme-Induced Epitope Retrieval (EIER)
  • Methods of Detection and Visualization
      • Substrates
      • Chromogens
      • 3,3’-diaminobenzidine (DAB) and 3-amino-9-ethylcarbazole (AEC)
      • Alkaline Phosphatase
      • Chromogen Enhancers
  • Immunohistochemical (IHC) Methods
      • Immunohistochemical (IHC) Techniques
      • Indirect Method
      • Direct Method
      • Avidin-Biotin Complex (ABC) Methods
      • Avidin-Biotin Complex (ABC) Method
      • Secondary Antibody
      • Peroxidase, Anti-Peroxidase (PAP) Method
  • Blocking Methods
      • General Blocking Methods
      • Avidin/Biotin Blocking
      • Serum Blocking
      • Blocking Endogenous Peroxidase Activity
      • Enzyme Blocking
  • Immunohistochemisty (IHC) Staining Reagents
      • General Reagents
      • Antibody Diluents
      • Buffers and Rinses
  • Immunohistochemical (IHC) Controls
      • Quality Control (QC)
      • Tissue Controls/Control Slides
      • Negative Quality Control (QC) Slides
      • Positive Quality Control (QC) Slides
      • System Quality Control (QC)
      • Antibody Validations
  • Troubleshooting
      • Troubleshooting
  • Record Keeping
      • Record Keeping
  • References
      • References

Additional Information

Level of instructruction: Basic to intermediate
Intended audience:  This program is designed as an educational and training tool for histology laboratory personnel. This course is also appropriate for students enrolled in clinical laboratory science and histology technician programs as a supplement to classroom instruction and clinical experiences.
Author information: Diane L. Sterchi, MS, HTL(ASCP) is a senior research associate for Covance Laboratories, Department of Discovery and Translational Services, Pathology in Greenfield, Indiana. She has worked in hospital, veterinary, and research settings in the discipline of histology. Ms. Sterchi holds an MS degree from Indiana Wesleyan University, Marion, Indiana. She has written several textbook chapters and manuals, is a frequent presenter at seminars and has conducted numerous workshops on topics including general histology, immunohistochemistry, and in situ hybridization.
Donna M. Hedger, BS, HT(ASCP) is a Pathology Manager at Gwinnett Hospital System in Lawrenceville, GA. She graduated from Georgia Southern University in Statesboro, Georgia with a Bachelor of Science degree in biology. She is a certified histotechnician. 
Reviewer information:  Tanisha N. Neely, HT(ASCP) is a freelance writer with a work history in histology, microbiology, and pharmaceutical research. Ms. Neely attended Indiana University where she earned a bachelors degree in arts & humanities from the School of General Studies and a Certificate in Histotechnology from the School of Medicine. She is a member of the National Society for Histotechnology and the Indiana Society for Histotechnology.
Course descripiton:  This course introduces the various techniques that are used in the preparation and evaluation of immunohistochemistry (IHC) slides. Procedures and terminology related to IHC are also discussed and strategies for troubleshooting problems are presented.


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