Intact skull chronic windows for mesoscopic wide-field imaging in awake mice

- Created bilateral chronic windows for imaging through the intact mouse skull.
- Includes a head-fixing screw for immobilizing the head during awake imaging.
- Preparation is stable for awake, widefield imaging of neuronal activity with GCaMP6.
- Functional connectivity maps were generated from awake spontaneous activity.

BackgroundCraniotomy-based window implants are commonly used for microscopic imaging, in head-fixed rodents, however their field of view is typically small and incompatible with mesoscopic functional mapping of cortex.New methodWe describe a reproducible and simple procedure for chronic through-bone wide-field imaging in awake head-fixed mice providing stable optical access for chronic imaging over large areas of the cortex for months.ResultsThe preparation is produced by applying clear-drying dental cement to the intact mouse skull, followed by a glass coverslip to create a partially transparent imaging surface. Surgery time takes about 30 min. A single set-screw provides a stable means of attachment (in relation to the measured lateral and axial resolution) for mesoscale assessment without obscuring the cortical field of view.Comparison with existing methodsWe demonstrate the utility of this method by showing seed-pixel functional connectivity maps generated from spontaneous cortical activity of GCAMP6 signals in both awake and anesthetized mice in longitudinal studies of up to 2 months in duration.ConclusionsWe propose that the intact skull preparation described here may be used for most longitudinal studies that do not require micron scale resolution and where cortical neural or vascular signals are recorded with intrinsic sensors or in transgenic mice expressing genetically encoded sensors of activity.