(A) Example of dual color image obtained by a GluA1 and GluA2 dual labeling (epifluorescence image on the left) and the super-resolved image acquired with d-STORM technique, both at 647 and 532 (middle and right image). GluA1 presents a HA tag, as shown on the scheme, HA tag is labelled with Alexa 647 and endogenous GluA2 with Alexa 532. An example of co-localization of two super-resolved clusters is reported down to the right. (B, C and D) represent the three steps necessary to determine super-resolved object co-localization (n = 6 cells, and 151 individual clusters). In (B), the size of an object is determined to discriminate potential clusters from the single particle, both having a different physiological meaning. In (C), the bivariate nearest neighbor distance is calculated to determine the average distance between two objects. Control distributions are obtained with the calculation of the distance when the same number of objects is randomly redistributed within the cell border. Finally, (D) is the distribution of Manders’ coefficients. No co-localization gives a null value, 100% co-localization a value of one. The dual labeling of GluA1/A2 reports the presence of small and large objects (B), distance between cluster of GluA1 labelled with anti-HA is closer to GluA2 cluster than randomly distributed (C) 95% of clusters co-localized at least partly (D).