What are the Steps Involved in Multiplex Staining?
The process of multiplex staining typically involves several key steps:
1. Antigen Retrieval: Ensures that the epitopes are accessible for antibody binding. Techniques such as heat-induced epitope retrieval (HIER) or enzymatic digestion are commonly used.
2. Blocking: Minimizes non-specific binding by using blocking agents such as serum or protein solutions.
3. Primary Antibody Incubation: Specific antibodies against the target antigens are applied.
4. Secondary Detection: Depending on the method, secondary antibodies conjugated to fluorophores, enzymes, or metal isotopes are used.
5. Visualization and Imaging: Fluorescent or chromogenic signals are captured using fluorescence microscopy, brightfield microscopy, or mass cytometry.
