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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 9/1/2020


  • 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.

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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


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