Figure 5.

Data simulations validate the ASA scoring system on large protein assemblies. (A) Application of the ASA scoring system to a simulated OAP distribution is given. A random fluorophore distribution is created (top left) and converted into both a binary image demonstrating pixels containing a fluorophore (left middle) and a simulated STED image (top right). The simulated STED image was placed through a series of experimental thresholds to test how well a threshold may identify fluorophore-containing pixels, compared to the known distribution of fluorophores (middle row). The ASA score for both known and calculated distributions was tallied (bottom row) and compared. (B) Sample AQP4 array data demonstrating the accuracy of various detection thresholds is given. (C) Quantification of the performance of the final ASA scoring algorithm using a 40% threshold on independent simulated data set is given.