Single Section Biomarker Measurement and Colocalization via a Novel Multiplexing Staining Technology

Measuring colocalization of multiple biomarkers may contribute to understanding tumor growth and progression. Traditionally, multiple biomarkers colocalization has been performed by processing multiple serial tissue sections. To provide true colocalization measures, we are investigating single section multiplexing techniques. Utilizing a fluorescent-based sequential stain and bleach system SSB we investigated multiplexing 8 markers in breast cancer tissue microarray sections. The experiments consisted of a 4 predictive biomarker panel ER, PR, HER2, and Ki67 and markers to assist in the image processing. The goals included comparing the immunofluorescent signal with bright field single chromogen IHC scores, the measurements of tumor heterogeneity, and to discover technical challenges. Early results suggest that SSB signals correspond to traditional IHC staining. Additionally, our work has highlighted improvements to workflow and the staining process. We present a review of our progress and expectations for this technology.