3D-Printed Droplet Microfluidic TRA-ICP-MS Platform: Single-Cell Analysis of Cadmium and Nanoplastics Coexposure in <i>Synechocystis</i>

Nanoplastics (NPs) in aquatic environments raise concerns as carriers that alter the bioavailability of co-occurring pollutants, such as cadmium (Cd), affecting combined toxicity. Precision toxicology now demands single-cell assessments to provide novel insights into pollutant interactions. In this study, we utilized a 3D-printed droplet microfluidic platform integrated with time-resolved analysis (TRA)─inductively coupled plasma mass spectrometry (ICP-MS)─to investigate the uptake behavior of single algal cells exposed to Cd and Eu-containing polystyrene (PS) NPs. 3D printing enables rapid prototyping and design flexibility for optimized microfluidic chips, while the monolithic structure eliminates assembly errors, reduces dead volume, and supports large-scale production. The droplet platform offers high-throughput single-cell encapsulation; coupled with TRA-ICP-MS, it minimizes cross-contamination and enhances sensitivity for multielement single-cell analysis. Single-cell analysis revealed that coexposure increased both the proportion of Eu/Cd-containing cells and the uptaken Eu/Cd content. The adsorption of Cd²⁺ imparted a more positive surface charge to PS NPs. This promoted heterogeneous aggregation between algal cells and PS NPs, thereby enhancing the bioavailability of PS/Cd²⁺ to the algae. Complementing these single-cell measurements, bulk-cell assays were conducted to evaluate the toxicological impacts of coexposure to Cd and PS NPs on microalgae. The results demonstrate that coexposure to PS NPs and Cd²⁺ resulted in synergistic effects, including enhanced growth inhibition, photosynthetic impairment, membrane damage, and increased secretion of extracellular polymers. These findings highlight the increased ecological risks posed to aquatic organisms by the coexposure to PS NPs and Cd²⁺, emphasizing the need for comprehensive assessments of nanoplastic-pollutant interactions in aquatic ecosystems.