Functional drug screening of tumor organoids on an Active-Matrix Digital Microfluidic Chip for cancer precision medicine

Abstract Cancer is a leading cause of morbidity and mortality worldwide, characterized by substantial inter-patient and intra-tumor heterogeneity that complicates treatment efficacy. While genomic profiling has advanced precision oncology, the prevalence of tumors lacking targetable mutations highlights the need for complementary functional assays. Here, we present a drug sensitivity testing platform based on active-matrix digital microfluidics (AM-DMF) integrated with patient-derived organoids (PDOs). We optimized protocols for sample loading, droplet generation and routing, and controlled drug exposure, achieving precise concentration gradients via iterative mix-split steps. By combining AM-DMF with PDOs, we enable automated liquid handling and high-content imaging to capture heterogeneous drug responses. This approach significantly enhances the efficiency, reproducibility, and sensitivity of drug testing-particularly for limited clinical samples and complex drug combinations. The platform provides an end-to-end workflow, from sample processing to quantitative response analysis, offering a systematic and scalable strategy for precision medicine. These findings establish an automatable, integrated pipeline for functional precision oncology, especially suited to scarce specimens and multi-drug regimens.