A high-throughput mass cytometry barcoding platform recapitulating the immune features for HCC detection

Early diagnosis of hepatocellular carcinoma (HCC), a highly malignant disease, is crucial for improving patient prognosis. Liquid biopsy has emerged as a promising approach for cancer detection, especially peripheral immunity profiling using mass cytometry time-of-flight (CyTOF). While the limitied throughput and high cost hamper its widespread application. To address these challenges, we developed an automated micro-droplet printing platform to achieve high-throughput mass cytometry barcoding (MCB) strategy. Utilizing a poly (methyl methacrylate) (PMMA) plane substrate, we established a multiplex barcoding checkboard in binary labeling strategy through "plate covering". To validate the utility of our system, we applied the multiplex barcoding system to label human peripheral blood samples from two cohorts consisting of 16 and 64 samples respectively, while successfully recapitulating the immune features associated with HCC. The ability of highthroughput barcoding samples empowers the application of CyTOF for large-scale HCC detection and other potential uses.