Fig. 3

Long-term high-resolution images of weak fluorescent protein-labelled tissue. A, B Image of YFP- and ChR2-YFP-labelled brain slices; the numbers in the images indicate the power of the laser used. Representative of n = 3 slices each conditioning. A is a large-field view and B is the image of the cortex (scale bars: 1 mm in A and 200 μm in B). C Representative images of high-resolution time-series imaging of ChR2-YPF-labelled tissue before and after AKS treatment. The numbers in the images indicate the power of the laser used. Representative of n = 3 slices each conditioning (scale bar: 20 μm). D Time-stack images showing the photobleaching effect before and after AKS treatment around imaging time of C. Representative of n = 3 slices each conditioning. E Analysis of the relative mean fluorescence intensity around time-series imaging of C (n = 3 slices each conditioning). All values are included in Additional file 11: Table S4. F 3D high-resolution imaging of the ChR2-YFP-labelled CRH neurons in the central amygdala. Representative of n = 2 samples. G Image cropped from F, showing how the dendrites of CRH neurons connect to the soma of CRH neurons in the central amygdala, indicated by a yellow arrow (scale bar: 2 μm). H Image cropped from F, showing the dendrites of CRH neurons connecting to dendrites of CRH neurons in the central amygdala, indicated by a yellow arrow (scale bar: 2 μm)