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.
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