Alignment of the different channels used for post-hoc ICC. Measured IR values for two presynaptic markers in each one of the individual ROIs: channel 1 (Alexa 488, anti-RIM1α in this case) and channel 2 (Alexa 594, anti-CB1R in this case) are shown in a.f.u (arbitrary fluorescence units) (A) or normalized to their mean IR value (B). Distribution of IR values of channel 1 (black bars) and channel 2 (red bars) in arbitrary fluorescence units (C) and normalized to mean levels (D). E) Graphical representation of the distribution of normalized IR values (<0.5 and >2 fold mean in blue and red, respectively) through the whole population of synaptic boutons. F) Normalized FM1-43 unloading kinetics of the three groups of synaptic boutons according to their RIM1α IR levels, means ± S.E.M. are plotted. Data from a single experiment (n = 671 boutons for the whole population trace; n = 96 boutons for IR levels >2, red trace and n = 231 boutons with IR levels < 0.5 mean value).
Ramírez-Franco et al. BMC Neuroscience 2013 14:127 doi:10.1186/1471-2202-14-127