Figure 3

Model validation. (a) A fluorescent liquid (dotted green area) between the coverslip and a divergent spherical lens was used as a known phantom sample to validate the model. (b) The estimated lens profile from MA-TIRF acquisitions follows the expected slope (dashed red line) of the lens within the observed region. (c) Color-coded depth representations of two independent acquisitions and reconstructions of the same biological sample (endothelial cells co-stained with Alexa-561- and Alexa-488-coupled phalloidin) reveal an accurate co-localization. White arrows mark adhesion sites. (d) The robust co-localization is further demonstrated in the 2D histogram reporting the relative depth between the two fluorophores, which exhibits an almost perfect diagonal pattern.