Fig. 7. Colocalization of integrins and stress fibers imaged by CLSF and TIRF microscopy. (A) HUVECs were fixed 30 minutes (a-c), 1 hour (d-f) and 6 hours (g-i) after plating. The left-hand column shows the images of integrin distribution (a,d,g); the centre column shows stress fiber distribution (b,e,h); the right-hand column shows superimposed images of the corresponding images in the left and centre columns (c,f,i). These images were taken 1 µm above the basal plane of the cell with a CLSF microscope. At 30 minutes after plating, integrins were colocalized with F-actin at the marginal region of the cell (arrows in c). Arrowheads in e and f indicate a typical stress fiber. At 1 hour after plating, the integrin clusters were located along the stress fibers (two arrows in d and f show typical cases). The same trend was observed in integrin cultures at 6 hours after plating (g-i). (B) HUVECs were fixed at 1 hour after plating and observed by TIRF microscopy. (a) A TIRF image of integrin clusters at FCs; (b) stress fibers; (c) superimposed images of a and b. Bright images of integrin clusters can be seen at the ends of stress fibers (arrows in a-c). Bar, 20 µm.
