4. “IHC is a biochemical assay technique that takes advantage
of chemical reactions to locate and visualize the interaction
between antibodies and their target antigen on sample
tissue”
What is Immunohistochemistry?What is Immunohistochemistry?
6. IHC SignificanceIHC Significance
While there are countless medical detection systems (x-ray, CT-
scan, MRI etc.); There are few identification tools available.
With new carcinogens being discovered and an ever increasing
population; the demand for cancer diagnostics is at an all-time high.
Even with the emergence of genetic testing; in many cases, IHC
remains the most cost-effective diagnostic tool from which
accurate and often life-saving diagnosis and/or prognosis can be
derived.
7. 2 Primary Applications2 Primary Applications
Diagnosis:
Identification of the
cause of an illness or set
of symptoms
{Infectious Agent?}
Prognosis:
Predicting the likely outcome of
an illness
8. Where is IHC Performed?Where is IHC Performed?
Clinical Labs
- Provide info on the health of the patient;
diagnosis/prognosis (Hospitals)
Reference Labs
- Large scale testing facility;
have extensive array of diagnostic
tools; receive toughest cases
Research labs
- Associated with
Universities; testing is
more experimental;
generate publications
14. ““Y” Use Antibodies?Y” Use Antibodies?
IHC takes advantage of the following antibody attributes:
Ability to locate and tag target proteins with incredible specificity
Constant region that can be located by secondary antibodies
Large molecules that can be outfitted with detection systems
19. 2. Histology (histo-)2. Histology (histo-)
Definition:
The study of the microscopic structure of organic
tissues. {Can you guess the tissue type depicted below?}
20. From Biopsy to SlideFrom Biopsy to Slide
Biopsy
Fixation (10% NBF)
Dehydration & Clearing
Embedding
Microtomy (3-4 microns)
Apply tissue to (+) charged slide
Baking (2-4 hours/overnight for fatty tissues)
22. 3. Chemistry3. Chemistry
Definition:
The science of the composition,
structure, properties and
reactions of matter
Examples of IHC Chemical Reactions:
Binding of antibody to antigen
Enzyme and chromogen interaction
41. CoverslippingCoverslipping
After being counterstained, slides are now ready to be
coverslipped to preserve the stain and allow for viewing under
the microscope
