Fig. 8. ICCM analysis of 5-YFP and -actinin-CFP. Two-photon fluorescence microscopy images collected at 37°C of -actinin-CFP (A-C) and 5-YFP (D-F) 2.5 hours after plating on 10 µg ml^–1 fibronectin. ICM and ICCM were conducted on each highlighted region of the cell. The distribution of -actinin, 5-integrin or colocalized species was determined via spatial autocorrelation or cross-correlation analysis. The dynamics of -actinin, 5-integrin or complexes moving together was determined via a temporal autocorrelation or cross-correlation analysis. Spatial functions were fit to Equation 4, and temporal functions were fit to one of Equations 11, 12 or 13. Equation 17 was used to estimate the fraction of each population that was immobile. The green bars show the fraction of a given species (diffusing, flowing or immobile) that is interacting with the second fluorescently tagged protein within a complex. The white part of the bar is the fraction noninteracting – i.e. the green bar in a region of the cell in Fig. 8A would represent the fraction of -actinin that is diffusing together with 5-integrin, whereas the green bar in Fig. 8D would represent the fraction of 5-integrin that is diffusing together with -actinin. Similarly, the green bar in Fig. 8B and 8E show the fractions of the respective proteins that are flowing together, and Fig. 8C and 8F represent the fraction of each protein population that is immobile and colocalized. Scale bars, 10 µm.