Focal adhesion dynamics and protrusion of the leading edge are critical for cancer cell migration and wound healing of epithelial cells. Massive actin polymerization at the leading edge generates a forward protrusion force at the cell membrane. At the same time, the entire actin network moves toward the cell center, which is called the retrograde actin flow. The interaction between the actin flows and the focal adhesions (FAs) has been proposed to enhance the membrane protrusion. However, roles of interaction between the retrograde actin flow and FAs largely remain unclear. In this project, we developed a new electroporation-based Single-Molecule Speckle (eSiMS) microscopy, which is easy-to-use live-cell fluorescent single-molecule technique with high spatiotemporal resolution (Yamashiro et al., Mol. Biol. Cell, 2014). Using eSiMS microscopy, we revealed that the retrograde actin flow in the front of mature FAs is fast and biased toward FAs, suggesting that FA exhibits "grabbing activity" towards the surrounding actin network, especially in the cell front. To further investigate roles of the grabbing activity of FAs on the retrograde flow, we examined molecular motions of FA components using SiMS microscopy. Interestingly, several FA components including vinculin, talin and integrinß1 exhibit biased motion toward FAs along the retrograde actin flow, suggesting that the flow recruits FA components to FAs for growth or maintenance. Furthermore, our single-molecule observations suggest that the grabbing activity of FAs on the retrograde flow is transmitted to the extracellular matrix (ECM), which may contribute to remodeling of ECM in front of FAs. The actin flow may remodel ECM at cell periphery during migration.