SignificanceWide-field optical imaging (WOI) can produce concurrent hemodynamic and cell-specific calcium recordings across the entire cerebral cortex in animal models. There have been multiple studies using WOI to image mouse models with various environmental or genetic manipulations to understand various diseases. Despite the utility of pursuing mouse WOI alongside human functional magnetic resonance imaging (fMRI), and the multitude of analysis toolboxes in the fMRI literature, there is not an available open-source, user-friendly data processing and statistical analysis toolbox for WOI data.AimTo assemble a MATLAB toolbox for processing WOI data, as described and adapted to combine techniques from multiple WOI groups and fMRI.ApproachWe outline our MATLAB toolbox on GitHub with multiple data analysis packages and translate a commonly used statistical approach from the fMRI literature to the WOI data. To illustrate the utility of our MATLAB toolbox, we demonstrate the ability of the processing and analysis framework to detect a well-established deficit in a mouse model of stroke and plot activation areas during an electrical paw stimulus experiment.ResultsOur processing toolbox and statistical methods isolate a somatosensory-based deficit 3 days following photothrombotic stroke and cleanly localize sensory stimulus activations.ConclusionsThe toolbox presented here details an open-source, user-friendly compilation of WOI processing tools with statistical methods to apply to any biological question investigated with WOI techniques.
Obtaining high signal to noise ratio is challenging in wide-field two photon microscopy and one must ensure the mouse brain can be imaged safely under high laser power. Here, we demonstrated a simultaneous thermal imaging and two photon imaging technique. The maximum temperature of the cortex was below 39°C using 400mW laser power with a 5 x 5mm field of view. Together with the brain activities under hind paw stimulation and at rest, we argued that high laser power for wide-field two-photon imaging can potentially be used while keeping the temperature under safety limit.
By paying careful attention to optical and mechanical design, a two-photon microscope (TPM) can image a large field of view (FOV) with high spatial and temporal resolution. Successfully applying such a TPM for functional neuroimaging requires careful consideration of the signal properties, physiological limitations, and image processing techniques. We present our thermal imaging results as well as functional imaging results in awake mice using a fast, large FOV TPM, and discuss how the system expands the spatial bandwidth available for studying functional connectivity.
Quick and accurate parcellation of neural networks has been a goal spanning multiple decades of research in the functional MRI world in order to organize and understand the overwhelmingly complex human brain with high statistical rigor. The same mathematical development in the mouse brain, which is frequently studied to understand human conditions, has been lagging. To this end, we perform high-throughput fluorescence imaging (GCaMP6f) in healthy mice in a cell-specific and non-invasive manner during rest and sensorimotor tasks in order to map healthy networks and understand patterns due to disease processes.
Modulation of brain state, e.g., by anesthesia, alters the correlation structure of spontaneous activity, especially in the delta band. This effect has largely been attributed to the ∼1 Hz slow oscillation that is characteristic of anesthesia and nonrapid eye movement (NREM) sleep. However, the effect of the slow oscillation on correlation structures and the spectral content of spontaneous activity across brain states (including NREM) has not been comprehensively examined. Further, discrepancies between activity dynamics observed with hemoglobin versus calcium (GCaMP6) imaging have not been reconciled. Lastly, whether the slow oscillation replaces functional connectivity (FC) patterns typical of the alert state, or superimposes on them, remains unclear. Here, we use wide-field calcium imaging to study spontaneous cortical activity in awake, anesthetized, and naturally sleeping mice. We find modest brain state-dependent changes in infraslow correlations but larger changes in GCaMP6 delta correlations. Principal component analysis of GCaMP6 sleep/anesthesia data in the delta band revealed that the slow oscillation is largely confined to the first three components. Removal of these components revealed a correlation structure strikingly similar to that observed during wake. These results indicate that, during NREM sleep/anesthesia, the slow oscillation superimposes onto a canonical FC architecture.
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