What are the Steps Involved in Immunohistochemistry?
The IHC process involves several steps:
Sample Preparation: Tissue samples are collected and fixed, usually in formalin, to preserve cellular structure. Sectioning: The preserved tissues are embedded in paraffin and then sliced into thin sections. Deparaffinization and Rehydration: Paraffin is removed, and the tissue sections are rehydrated. Antigen Retrieval: Techniques like heat-induced epitope retrieval (HIER) are employed to unmask antigens. Blocking: Non-specific binding sites are blocked to minimize background staining. Primary Antibody Incubation: The tissue sections are incubated with primary antibodies specific to the target antigen. Secondary Antibody Incubation: Secondary antibodies conjugated with enzymes or fluorophores bind to the primary antibodies. Visualization: The enzyme or fluorophore is activated to produce a visible signal, indicating the presence of the target antigen.
