Geometry-dependent migration dynamics of fibroblasts on ECM protein micropatterns

Extracellular matrix (ECM) micropatterns provide well-defined adhesive environments for investigating how geometric confinement influences cell migration. Here, we combined ECM protein micropatterning with quantitative live-cell microscopy to examine NIH 3T3 fibroblast migration on straight and sinusoidal adhesive tracks. Time-lapse bright-field imaging, actin fluorescence imaging, centroid tracking, migration-rate quantification and aspect-ratio analysis revealed distinct migration behaviours under confined adhesion. On straight tracks, cells exhibited either oscillatory migration, characterised by repeated cycles of protrusion, retraction, and migration-rate bursts, or confined migration, characterised by limited centroid displacement and slow morphological remodelling. Sinusoidal tracks further introduced curvature-associated changes in migration dynamics, with cells adjusting leading-edge orientation along the curved path or becoming locally confined within curved regions. These observations demonstrate that adhesive geometry influences cell migration beyond simple path guidance and highlight the value of engineered ECM micropatterns combined with live-cell microscopy for resolving dynamic migration behaviours under confined conditions.