Figure 7
Visualization, tracking and analysis of the dimerization of single SNAP-tagged D2L receptors and wild-type D2L receptors labeled with the fluorescent antagonist 1c.
(a,d) Schematic representation of the constructs. (b,e) Representative images of single CHO cells stably transfected with the two constructs, labeled and visualized by TIRF-M. Spot densities were 0.69 spots μm−2 (b) and 0.64 spots μm−2 (e). Inserts correspond to higher magnification images of the areas in the white boxes. Scale bar, 10 μm. (c,f) Representative intensity distributions of fluorescent spots identified over the first 10-frame time window of TIRF illumination of CHO cells, stably transfected with the corresponding constructs and labeled with the fluorescent ligand 1c (300 nM). Data were fitted with a mixed Gaussian model. A mixed Gaussian fit after partial photobleaching (dotted line) was used to estimate precisely the intensity of a single fluorescent spot in each image sequence.
