Figure 7. Use of HER4 binding Affimers in super-resolution imaging and single molecule tracking.
(A) Phage ELISA for HER4 binding Affimers. (B) Average photon counts/pixel for HER4-binding Affimer labelled with CF640R and bound to CHO cells transfected with HER4 and to MCF7 cells expressing endogenous levels of HER4. (C) Wide field image of CHO cells transfected with HER4-CYT-eGFP showing localisation of HER4 via GFP fluorescence (top) and labelled with HER Affimer–Alexa647 (middle). The corresponding dSTORM image of HER4 Affimer conjugated to Alexa647 (bottom) with a 25 nm localisation precision. Scale bar = 2 μm. Right plots to show the number of molecules and cluster size of clusters identified by dSTORM. (D) Diffusion coefficients (left panel), and MSD curve (right panel) of HER4 Affimers labelled with CF640R and tracked on MCF7 cells expressing endogenous HER4.