Reconstruction of three-dimensional spatial patterns of rat cerebellar cortex activity in vivo using voltage sensitive dyes and optical sectioning

The three-dimensional neuronal activity of parallel fibers and Purkinje cells in rat cerebellar cortex is investigated in vivo. Using a voltage-sensitive dye (RH795) and imaging under an epi-fluorescence microscope and a CCD video camera, the response of cerebellar neurons to electrical stimulation of parallel fibers is studied. A deblurring algorithm is used on serial sectioned images to refine the spatial pattern of optical activity as well as to reveal the underlying neuronal events. This method reduces the error from out-of-focus components, improving the quantification and mapping of neuronal activity at low magnification