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Updated: Apr 17, 2026
Multiparametric Tumor Organoid Drug Screening Using Widefield Live-Cell Imaging for Bulk and Single-Organoid Analysis
Published on: December 23, 2022
Functional drug screening of tumor organoids on an active-matrix digital microfluidic chip for cancer precision
Ruimeng Sun1,2, Zhengyang Feng3, Tong Wu1,2
1School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230026, China.
Abstract:
Cancer is a leading global cause of morbidity and mortality, with inter- and intra-tumor heterogeneity contributing to suboptimal therapeutic outcomes. While genomic profiling has advanced precision oncology, many tumors lack targetable mutations, underscoring the need for functional assays. Here, we present a drug sensitivity testing platform for patient-derived organoids (PDOs) based on active-matrix digital microfluidics (AM-DMF). The system comprises a microfluidic timing-control board (<50 ms switching precision), a temperature-regulation module (37 ± 0.3 °C) and an integrated imaging module for multiscale visualization. Droplet generation, transport, mixing, and dilution on the AM-DMF platform are programmably controlled via electrowetting-on-dielectric (EWOD) actuation, with volume stability from 1 to 100 nL per droplet (CV < 3%). A multilayer design of the AM-DMF chips suppressed crosstalk and leakage current, enabling stable operation over 10 h. PDO-loaded droplets were efficiently generated through symmetric "one-to-two" splitting, achieving >71% success within 4-5 tearing cycles. Evaluation of cisplatin cytotoxicity across multiple PDOs on AM-DMF chips revealed time- and dose-dependent responses, including early morphological contraction (12 h) and subsequent structural disintegration (72 h) at high concentrations. Live/dead staining confirmed membrane damage under high-dose treatment, reflected by increased red fluorescence. Quantitative analysis demonstrated consistent dose-dependent inhibition of viability across all PDOs. Notably, on-chip measurements showed response trends consistent with conventional 96-well plate assays, yet with moderately higher inhibition rates, indicating enhanced assay sensitivity. Overall, we have developed an automated, high-throughput drug-testing system for PDOs, wherein the entire workflow, including organoid culture, precise drug concentration gradient generation, multi-dose drug screening, staining, and imaging, was seamlessly integrated and performed on a single active-matrix digital microfluidics chip. Collectively, this integrated AM-DMF platform offers a systematic, scalable strategy for functional precision medicine, especially suited to scarce clinical specimens and complex multidrug regimens.
Insights
We developed a novel drug sensitivity testing platform using active-matrix digital microfluidics (AM-DMF) for patient-derived organoids (PDOs). This high-throughput system enables precise drug screening for personalized cancer therapy.
Area of Science:
- Biotechnology and Biomedical Engineering
- Cancer Research and Precision Oncology
Background:
- Cancer heterogeneity poses challenges to effective treatment, driving the need for functional drug sensitivity assays beyond genomic profiling.
- Patient-derived organoids (PDOs) offer a promising model for personalized cancer therapy but require efficient drug screening platforms.
Purpose of the Study:
- To develop and validate an automated, high-throughput drug sensitivity testing platform for PDOs utilizing active-matrix digital microfluidics (AM-DMF).
- To integrate organoid culture, drug gradient generation, screening, staining, and imaging onto a single microfluidic chip for functional precision medicine.
Main Methods:
- Development of an AM-DMF system with precise timing control, temperature regulation, and integrated imaging.
- Programmable droplet generation, transport, mixing, and dilution using electrowetting-on-dielectric (EWOD) actuation for PDO handling.
- Evaluation of cisplatin cytotoxicity on PDOs using the AM-DMF platform, including morphological and viability assessments.
Main Results:
- The AM-DMF platform demonstrated stable operation with precise droplet volume control (1-100 nL, CV < 3%) and efficient PDO droplet generation (>71% success).
- On-chip drug screening revealed time- and dose-dependent cytotoxic effects of cisplatin on PDOs, consistent with conventional assays but with potentially higher sensitivity.
- Integrated workflow from organoid culture to multi-dose drug screening and imaging was successfully demonstrated on a single chip.
Conclusions:
- The integrated AM-DMF platform provides a systematic and scalable solution for high-throughput drug sensitivity testing of PDOs.
- This technology facilitates functional precision medicine, particularly for scarce clinical specimens and complex drug regimens, improving therapeutic outcomes.
