The IHC process typically involves the following steps: 1. Fixation: Tissue samples are preserved using chemicals such as formalin. 2. Embedding: Fixed tissues are embedded in paraffin to create solid blocks. 3. Sectioning: Thin tissue sections are cut from the paraffin blocks and mounted on slides. 4. Deparaffinization and Rehydration: Paraffin is removed, and tissues are rehydrated. 5. Antigen Retrieval: Heat or enzymatic treatment is used to expose antigenic sites. 6. Blocking: Non-specific binding sites are blocked. 7. Primary Antibody Incubation: Slides are incubated with primary antibodies specific to the target antigen. 8. Secondary Antibody Incubation: Slides are incubated with enzyme or fluorophore-conjugated secondary antibodies. 9. Visualization: Substrate is added if using enzyme-conjugated antibodies, or slides are directly observed under a fluorescent microscope if using fluorophore-conjugated antibodies.
