Understanding the basis of brain function requires knowledge of cortical operations over wide-spatial scales, but also within the context of single neurons. In vivo, wide-field GCaMP imaging and sub-cortical/cortical cellular electrophysiology were used in mice to investigate relationships between spontaneous single neuron spiking and mesoscopic cortical activity. We make use of a rich set of cortical activity motifs that are present in spontaneous activity in anesthetized and awake animals. A mesoscale spike-triggered averaging procedure allowed the identification of motifs that are preferentially linked to individual spiking neurons by employing genetically targeted indicators of neuronal activity. Thalamic neurons predicted and reported specific cycles of wide-scale cortical inhibition/excitation. In contrast, spike-triggered maps derived from single cortical neurons yielded spatio-temporal maps expected for regional cortical consensus function. This approach can define network relationships between any point source of neuronal spiking and mesoscale cortical maps.DOI: http://dx.doi.org/10.7554/eLife.19976.001

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 widefield 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 30 minutes. A single set-screw provides a stable means of attachment 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.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.

Based on EEG data recorded from 11 subjects with eyes open and the left mastoid (M) reference, three data sets were generated by re-referencing to the conventional linked mastoids (L), average (A) and the new 'infinity' (I) reference provided by the reference electrode standardization technique (REST, Yao 2001 Physiol. Meas. 22 693-711). The EEG power in the alpha frequency band with the four different references was calculated and compared with respect to the total energy and spatial amplitude weight centre (AWC) coordinates, to compare the effects of different references on power mapping in the frequency domain. Compared with the I reference, the AWCs of the EEG with the M reference show significant shifts to the right, frontal and superficial positions, the L reference significant shifts to frontal and superficial positions, and the A reference shifts the AWC significantly to a deeper position. Furthermore, the power maps of the M and L references have larger total power than the I reference, while that of the A reference has the smallest total power. These results confirm that different choices of reference electrodes result in systematic changes in the distribution of EEG frequency power, and in order to reduce the effect of such systematic shifts on the explanation of EEG mappings, a common reference is necessary for EEG research. We recommend the I reference for objective use in cross-laboratory studies and clinical practices, as it is far from all the other electrodes and can act as a neutral reference.

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