We demonstrate the experimental setup and the use of multispectral fiber-photometry during fMRI in vivo for concurrent measurements of GCaMP6f, CBV, and BOLD. The fluorescent intensity of GCaMP6f and Rhodamine B, a red fluorescent dye which represents CBV, increased during spontaneous motion in the freely moving animal. GCaMP6f, CBV, and BOLD increased when forepaw electric stimulation was given in the simultaneous fMRI and fiber-photometry. We conclude that simultaneous multispectral fiber-photometry and functional MRI is a powerful tool which has an advantage in both temporal and spatial dimensions. Rhodamine B could be a surrogate CBV indicator in the regions of interest.
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