What are the Key Steps in Chromogenic Multiplex Staining?
1. Tissue Preparation: Sections are prepared from paraffin-embedded or frozen tissue samples. 2. Antigen Retrieval: Heat-induced epitope retrieval (HIER) or enzymatic digestion is used to unmask antigens. 3. Blocking: Non-specific binding sites are blocked to prevent background staining. 4. Primary Antibody Incubation: Specific primary antibodies are applied to the tissue. 5. Secondary Antibody Application: Enzyme-conjugated secondary antibodies are used to detect primary antibodies. 6. Chromogenic Substrate Addition: Substrates are added to visualize the staining. 7. Counterstaining: Hematoxylin or other dyes are used to provide contrast.
