This document provides an overview of an advanced histopathology techniques course. The course covers topics related to immunohistochemistry (IHC), immunofourescent methods, enzyme histochemistry, and in-situ hybridization. IHC is described as the localization of antigens in tissue sections using labeled antibodies and visualized with markers. Key steps in IHC include sample preparation, antigen retrieval, blocking background staining, antibody incubation and detection, and counterstaining. IHC is useful as a diagnostic tool, for assessing disease progression and cancer treatment, and differentiating cell types.
8. IHC takes its name from the roots "immuno," in
reference to antibodies used in the procedure,
and "histo," meaning tissue (compare to
immunocytochemistry). The procedure was
conceptualized and first implemented by Dr.
Albert Coons in 1941
Definition
IHC is a method for demonstrating the presence
and location of (protein) cellular or tissue
constituent (as antigens), by mean of antigen
9. How
Immunohistochemistry is the localization of
antigens in tissue sections by the use of labeled
antibody as specific reagents through antigen-
antibody interactions that are visualized by a
marker such as enzymes.
10. It is accomplished with antibodies that recognise
the target protein .and this Ab bind only to the
protein of interest in the tissue section
The aim is to achieve reproducible and
consistent demonstration of antigens with the
minimum background staining whilst
preserving the integrity of tissue architecture.
11. For optimal staining to occur it is necessary to
use the primary specific antibody at the correct
dilution
Blocking background staining (The major
causes of background staining in
immunohistochemistry are ionic interactions
and endogenous enzyme activity and
unbounded antibodies)…Endogenous enzyme
block – protein blocking -washing
Unmasking of antigen ……antigen retrieval
Electrostatically charged slide
12. Incubation
to prevent evaporation of antibodies, incubations
must be carried out in a moist atmosphere.
Using the wax pen around the section to assist
with retaining the reagents on the section
13. steps
-Preparation of sample (retrivation)
-Immunohistochemical reaction
-Visualizing the reaction
-Counter stain.
14.
15. Applications of IHC
especially advantages as a diagnostic tool for
infectious disease
It is useful for assessing the progression and
treatment of disease such as cancers
Differentiate between types of malignant cells
Detection of metastatic tumours