Real-Time Label-Free Imaging and Quantitative Analysis of Macrophage Morphodynamics Using Optical Diffraction...

This study establishes a label-free optical diffraction tomography (ODT)-based workflow for live-cell imaging and analysis to observe time-dependent morphological changes in macrophages. The method enables continuous recording of single-cell morphology and movement over extended periods under stable environmental conditions and allows extraction of quantitative parameters, including projected area, perimeter, and average migration speed. This workflow provides a practical approach for capturing dynamic cellular behaviors at the single-cell level without exogenous labeling. Using RAW264.7 macrophages as a model, time-lapse imaging was performed under lipopolysaccharide stimulation with baicalin pretreatment to capture dynamic cellular changes under different conditions. Representative cells were selected for tracking and quantitative analysis. The results show that this workflow supports stable long-term single-cell tracking and reflects temporal changes in cell morphology and motility. This approach provides a label-free method for observing dynamic cellular behaviors in response to different stimuli and can serve as a useful complement to conventional endpoint-based assays. It may also be applicable to other adherent cell types for studies of cell morphodynamics.