Morphological response of HeLa cells to Doxorubicin exposure in a microfluidic environment.

Cervical cancer remains one of the most frequently diagnosed malignancies among women worldwide. Therefore, analysing cell morphology is essential for advancing our understanding. In this study, we demonstrate that our microfluidic device generates stable laminar flow, establishing it as a reliable platform to evaluate the dose-dependent response of HeLa cells to doxorubicin (DOX; 0.1–3 µM). Propidium iodide staining was utilized to quantify cell viability and automated image analysis was performed through three methods (Fiji, QuPath and GEMA algorithm). Exposure to low DOX concentrations (0.1–0.5 µM) exerted a predominantly cytostatic effect, limiting cell expansion and mitigating density-dependent confluent stress. Conversely, higher concentrations (1–3 µM) induced severe, dose-dependent cytotoxicity. Crucially, spatial confinement within the microfluidic channel geometries promoted the formation of cell aggregates that exhibited enhanced localized resistance to DOX, highlighting the platform’s utility for resolving microenvironment-mediated drug responses