Quantitative Analysis of Single Cell Adhesion and Migration Using μ-PIV and TIRF

In order to quantify the single cell adhesion and migration, we propose a novel method using total internal reflection florescence (TIRF) microscopy and μ-PIV. A living cell has focal contacts on the substrate so that cell adhesion depends upon the formation of adhesive contacts between the cell and the substrate. Determination of the cell-substrate contact area (or the number of contact points) is necessary in order to understand how biomaterial properties influence cell adhesion. By using TIRF microscopy, we can obtain the image of near-focal contact (sub-micrometer scale). From the obtained images, it is determined whether the cell adheres properly. The cell movement which is caused by changing the hydrodynamic condition is also observed. The moving focal contacts are measured quantitatively with μ-PIV. This combination of TIRFM and μ-PIV proves to be very useful for analyzing cell viability in a new environment.