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Background: Craniotomy-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 Method: We 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., Results: The 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 30min. 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 Methods: We 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., Conclusions: We 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., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The extraction and detection of specific responses from a large amount of background noise has been the subject of a considerable body of research in brain functional imaging, and more specifically in optical intrinsic signal imaging. Recent work by Kalatsky and Stryker (2003) showed that by combining different conditions and using periodic stimuli, recording times can be reduced. Spectral decomposition is then used to provide amplitude and phase information locked to the stimulus. A drawback of the above method is that by focusing only on a single harmonic, response information is limited. The shape of the hemodynamic response function (HRF) and the temporal variations in the neural responses cannot be assessed. In this work it is argued that additional information about neural responses can be gathered by using higher harmonics. Moving bars were used to generate visuotopic maps on large portions of the cat visual cortex. Up to four simultaneously bars moving repetitively across the visual field at different frequencies were used to sample the HRF in the Fourier domain. The HRF profile obtained with continuous stimulation was spatially homogeneous throughout the cortex and similar to the HRF profile obtained using episodic stimulation. Furthermore, by modeling the optical response as a convolution between HRF and neuronal responses, the ratio of the second harmonic to the first provided an estimation of the receptive field size. This was further validated by measuring spatial frequency selectivity. Therefore, the use of higher harmonics opens new avenues to estimate receptive field size from temporal signals., ((c) 2010 Elsevier B.V. All rights reserved.)