Supplementary Figure 9: In vivo imaging of glutamate response of thalamocortical axons in vS1 cortex during vibrissa stimulation with full AO.

(a) Coronal section of the mouse brain shows the projection from VPM thalamus to vS1 cortex labeled by SF-Venus-iGluSnFR.A184S. (b) In vivo two-photon imaging of the thalamocortical axons in layer 4 of vS1 cortex is shown as the representative of 4 mice. Red box indicates the imaging region shown in panel d. (c) Air puff whisker stimulation during two-photon imaging. (d) Two-photon imaging of SF-Venus-iGluSnFR.A184S shows the glutamate release from thalamocortical axons and boutons varies as a function of depth as the representative of 7 measurements across 4 mice. Glutamate responses are averaged over the ROI defined by the red box region for all depths. (e, f) Comparison between System AO and Full AO correction on functional imaging of thalamocortical axons and boutons that express SF-Venus-iGluSnFR.A184S at 350 µm below the pia. Corresponding glutamate responses to vibrissa stimulation in the ROIs defined by the dashed circles are shown in panel f. (g, h) Same as panels e and f but at the depth of 650 µm. Experiments in panels e-h were repeated in 2 mice.