Development of a 3-D Physical Dynamics Monitoring System Using OCM with DVC for Quantification of Sprouting...

The extracellular matrix (ECM) plays a key role during cell migration, proliferation, and differentiation by providing adhesion sites and serving as a physical scaffold. Elucidating the interaction between the cell and ECM can reveal the underlying mechanisms of cellular behavior that are currently unclear. Analysis of the deformation of the ECM due to cell-matrix interactions requires microscopic, three-dimensional (3-D) imaging methods, such as confocal microscopy and second-harmonic generation microscopy, which are currently limited by phototoxicity and bleaching as a result of the point-scanning approach. In this study, we suggest the use of optical coherence microscopy (OCM) as a live-cell, volumetric, fast imaging tool for analyzing the deformation of fibrous ECM. We optimized such OCM parameters as the sampling rate to obtain images of the best quality that meet the requirements for robust digital volume correlation (DVC) analysis. Visualization and analysis of the mechanical interaction between collagen ECM and human umbilical vein endothelial cells (HUVECs) show that cellular adhesion during protrusion can be analyzed and quantified. The advantages of OCM, such as fine isotropic spatial resolution, fast time resolution, and low phototoxicity, make it the ideal optic tool for 3-D traction force microscopy.