Tissue Preparation: Tissue samples are fixed and embedded in paraffin before being sectioned onto slides. Antibody Staining: Multiple primary antibodies specific to the desired biomarkers are applied. Each antibody is conjugated to a unique fluorophore or chromogen. Detection: Visualization is achieved through fluorescence or chromogenic detection, enabling the identification of multiple targets within the same tissue section. Image Analysis: Advanced software is used to analyze the stained tissue, quantifying the presence and distribution of the biomarkers.
