Real-Time Three-Dimensional Imaging of Cell-Substrate Adhesion Structures Based on Plasmonic Scattering Microscopy

Real-time monitoring of cell-substrate interactions remains a significant challenge in cellular biomechanics. Here, a plasmonic scattering microscopy (PSM) integrated with three-dimensional image reconstruction is presented for real-time analysis of the cell-substrate interface, simultaneously achieving submicrometer spatial resolution. The system was validated by imaging microspheres with known geometries and achieved a mean coefficient of determination of 86% in the reconstructed images. Furthermore, three-dimensional mapping of the interface between MCF-7 cells and a Ag substrate revealed three distinct contact modes, with the dynamic changes in cell adhesion gap ranges for each mode in agreement with previous predictions. This study provides insights into achieving label-free, real-time monitoring of the three-dimensional dynamic evolution of cell-substrate adhesion structures and demonstrates that the technique has the potential to serve as a valuable tool for analyzing cell adhesion dynamics.