Combinations of immunology, histology and chemistry techniques

1. IMMUNOHISTOCHEMICAL
(IHC)
By:
1. Farina Bt Nor Hashimi (12171211079)
2. Nurul Fateha Wathiqah Bt Johari (12171211057)
3. Siti Hawa Bt Rahim (12171211124)

2. Immunohistochemistry
What is it?
- It’s a combination of immunological, histological and biochemical technique for the
identification and visualization of antigens in cells and tissues by using antibodies.
- It gives spatial location, can be used to locate particular cells and proteins and also
can be used to identify cellular events – e.g. apoptosis

3. ̵ Although less sensitive quantitatively than immunoassays such as western blot or
ELISA, it enables the observation of processes in the context of intact tissue.
Importance of IHC technique:
- Can be detected by several ways
- Bind to antigen in specific manner
- Antibodies can be raised against almost any type of antigen

4. Principle
◦ Wide used biological technique
◦ Develop from antigen-antibody binding reaction
◦ Method that visualize distribution and localization of specific antigen or
cellular component in separated tissue or tissue section
◦ Provide in situ information

5. 1. Primary antibody binds to specific antigen
2. Antibody-antigen complex is formed by incubation with a secondary, enzyme
conjugated and antibody
3. With presence of substrate and chromogen, the enzyme catalyzes to generate
colored deposits at the sites of antibody-antigen binding

6. IHC APPLICATIONS
i) Disease diagnosis
-To diagnosed a cancer as benign or malignant
-To determine the stage and grade of a tumor
-To identify the cell type and origin of a
metastasis to find the site of the primary tumor
Example: Diagnostic of breast cancer, lung
cancer, and prostate cancer.

7. ii) Drug discovery and development
Discovery including Drug Efficacy and Safety
Studies, Drug Repositioning, Patient
Stratification, Target Validation and Biomarker
Expression Studies.
Development to test drug efficacy by detecting
either the activity or the up- or down regulation of disease targets.

8. Case Study
◦ Example : Breast Cancer
◦ A patient in early forty’s was diagnosed with tumor in breast by
using mammogram.
◦ A sample of the tumor cell was taken via biopsy to examined the
suspicious area for a possibility of cancer cell.
◦ IHC technique was conduct to determine the cellular activity.
◦ In this case, IHC is use to differentiate the malignancy or benign of
the cell

9. ◦ To detect the cell condition, some steps of IHC was applied.
1. preparation of antibody
-monoclonal or polyclonal
2. preparation of tissue
-usually use aldehyde or frozen
3. antibody application
-two ways (direct or indirect)
4. visualization of antibody
-using immunofluorescence
-fluorescence/staining tag was labeled on antibody

10. Example of breast cancer cells using Immunohistochemical
technique. The cells are counterstained with hematoxylin.