Figure 3
Single and double immunolabeling experiments on non-contrasted thin sections. (a–b) Immunogold synaptic protein localization on uncoated Tokuyasu sections. (a) Cerebellar synapses identified by the synaptic cleft (arrows), round dendritic spines (s) and the presence of many packed synaptic vesicles (sv) labeled with gold nanoparticles (white dots) for VGLUT1. (b) Bassoon immunopositive gold particles (white dots) are located at close proximity of the synaptic cleft (arrowheads) at the presynaptic side (sv). (c) Correlative confocal laser scanning microscopy and scanning electron microscopy shows the expression of the presynaptic protein VGLUT1 (green) and postsynaptic protein calbindin (magenta) in several cerebellar synapses (arrows). 170 nm fiducials (in image these appear as two round light cyan spheres) are used to align the light and electron microscopy images. (d) Many synaptic vesicles (sv) in a parallel fiber synaptic bouton stained with VGLUT1 (green) make a synaptic contact (arrow) with a dendritic spine (s) labeled with calbindin (magenta). (e) Super-resolution synaptic protein localization on non-contrasted Tokuyasu cerebellar sections. Synapse identified by the synaptic cleft (arrow) and the presence of many packed synaptic vesicles (sv) and a dendritic spine (s). Bassoon immunopositive signal (red) is located at close proximity of the synaptic cleft (arrow) at the presynaptic side (sv). (f) Dual colour GSDIM image of Tom20 (magenta) appearing at the outer membrane (arrows) of a mitochondria and the expression of ATP-Synthase (green) in the inner part of the mitochondria (asterisk) of Cos7 cells. All electron microscope images were acquired with a scanning electron microscope using an in-lens detector at an acceleration voltage of 1.5 keV. Scale bars: 100 nm.
