Recording functional signals from various brain regions is crucial in many biomedical studies. For example, monitoring multiple brain regions such as the cortex, the thalamus, and the striatum is necessary under general anesthesia. However, it is still challenging to achieve cortical-wide field-of-view, deep penetrating, high resolution and high recording speed simultaneously. In this study, we proposed the use of epifluorescent microscopy to simultaneously record neuronal activities in the superficial cortex and functional signals from multiple deep brain regions. Neural activity intensity in deep brain regions was relayed to the superficial cortex via optical fibers, with their depths adjusted according to the thickness of cover glass. We have demonstrated the capability to simultaneously record the entire superficial cortical neuronal activities and functional signals of the thalamus and the substantia nigra (SN) during the anesthesia and recovery processes. This method can be integrated with other advanced imaging techniques such as miniscopes and may prove useful in many other applications.
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