Fig. 4

Monitoring Ca²⁺ activities in awake head-fixed mice. a Wide-field image of a Thy1-GCaMP6f mouse implanted with a See-Shell taken 6 weeks after implantation. Four locations indicated by the colored blocks were imaged using a 2P microscope. Scale bar indicates 1 mm. b Schematic of the mouse on the custom-designed disk treadmill used for 2P imaging. c Average intensity images calculated from 5-min time series acquired 200–300 µm deep from pial surface. Scale bar indicates 100 µm. d Color-coded time series of Ca²⁺ activities of neurons identified and annotated by open circles in the respective average intensity images in c. e Time lapse Ca²⁺ imaging: Left: Average intensity images calculated from 5 min time series acquired from the same field of view in a See-Shell-implanted Thy1-GCaMP6f mouse in six separate imaging sessions. Activities of individual neurons, indicated by arrows, are shown in the right. Scale bar indicates 100 µm