The procedure for multiplex staining typically involves several key steps:
Tissue Preparation: Sections of tissue are prepared and placed on slides. Antigen Retrieval: Methods like heat-induced epitope retrieval (HIER) or enzyme digestion are used to unmask antigens, making them accessible to antibodies. Blocking: Non-specific binding sites are blocked to reduce background staining. Primary Antibody Incubation: Primary antibodies specific to the targets of interest are applied. Secondary Antibody Incubation: Secondary antibodies conjugated with fluorophores or enzymes are used to detect the primary antibodies. Visualization: The stained tissue section is examined using a microscope equipped with appropriate filters or light sources for detecting the different labels.
