Prerequisite/Corequisite Learning:
- Discuss/define terminology and basic principles of Ag-Ab reactions
- Discuss/define terminology and basic principles of enzyme reactions
- Describe the immune system and its cells/tissue/organs
- Recognize Hematoxylin and Eosin (H&E) stained normal structure of the immune system using light microscope
- Recognize H&E stained normal tissue structure using light microscope
- Describe the structure of antibodies and types of antibodies
- Define antigenic determinants
- Distinguish between monoclonal and polyclonal antibodies
- Describe basic principles of tissue/cell fixation and preservation; tissue processing and microtomy
- Describe basic theories of stains and dyes
Learning Outcomes:
- Identify the use and purpose for performing immunohistochemistry staining
- Define/describe the immunohistochemistry principles and mechanisms underlying Ag- Ab reactions:
- Single step methods
- Multiple step methods
- Peroxidase anti-peroxidase (PAP) method
- Avidin-biotin (ABC) and labelled streptavidin biotin (LSAB) methods
- Polymer method
- Identify enzyme reactions in immunohistochemistry
- Discuss and compare visualization and amplification methods:
- Identify use of Fluorescent dyes
- Identify use of chromogenic dyes-Enzyme/Chromogen systems:
- Peroxidase and DAB (diaminobenzidine) or AEC (aminoethylcarbazole)
- Phosphatase and fast red
- Describe the limitations of IHC staining:
- Define and describe antigen preservation
- Recognize and describe antigen localization
- Discuss optimal fixation methods
- Identify antigen specificity
- Describe non-specific/specific background staining and potential causes
- Identify use of blocking methods to increase Ag-Ab binding specificity
- Identify antibody availability and differentiate between:
- Monoclonal antibodies
- Polyclonal antibodies
Author: Masaye Tanaka, ART, BSc, MEd
Version Date: September 2019
PEP hours: 16.5
CPS credits: 1.6
*Note: PEP hours and/or CPS credits will only be awarded upon successful completion of Final Exam.
