Figure S1 - Synapto-Protective Drugs Evaluation in Reconstructed Neuronal Network

Automated quantification of synapses with SynD software. Synaptic disconnection was assessed by counting v-GLUT1 clusters affixed to MAP-2 positive striatal dendrites. Images were all obtained with the same acquisition parameters and were similarly processed with ImageJ software before being used for quantification. (a) v-GLUT1/MAP-2 merge used for quantification using SynD software.(b) Automatic soma detected using thresholding followed by morphological opening and closing. (c) Dendrites detected using steerable filters. A cost function evaluates each pixel around the current neurite mask, and those with a cost lower than a specified threshold are added to the mask. The cost function takes into account how neurite-like the neighborhood around the candidate pixel is (i.e. ideally a bright line segment surrounded by dark regions on either side), and if the candidate pixel is in the direction of the already detected parts of the neurite. (d) The detected neurite mask overlaid on the v-GLUT1 channel, only the parts located either within the neurite mask or a small neighborhood around it are used for synapse detection. (e) Synapse regions are detected by thresholding the v-GLUT1 channel; putative centers are located by deconvoluting the image. (EPS)