Fixation: Preserving the tissue to maintain cellular structure and antigenicity. Embedding: Embedding the fixed tissue in paraffin wax to facilitate sectioning. Sectioning: Cutting thin slices of the tissue for mounting on slides. Deparaffinization and Rehydration: Removing the paraffin and rehydrating the tissue sections. Antigen Retrieval: Unmasking antigens to enhance antibody binding, often through heat-induced methods. Blocking: Preventing non-specific binding of antibodies. Primary Antibody Incubation: Applying the primary antibody that binds to the antigen of interest. Secondary Antibody Incubation: Applying a secondary antibody that binds to the primary antibody and is linked to a detectable marker. Detection: Visualizing the antigen-antibody complex using appropriate substrates or fluorescent dyes.
