Figure 1.

Methods for obtaining voltage-sensitive dye recordings from Purkinje cell axons. Schematic of the experimental setup shown in the middle: IR-DIC video microscopy was used for patching neurons; the voltage-sensitive dye was excited using a 532 nm solid-state laser in wide-field illumination mode; emission light was recorded with a high-speed CCD-camera; a spinning-disc confocal scanner was used for morphological reconstruction. A, A composite EGFP fluorescence image of a Purkinje cell with intact axonal arbor generated from stacks of images obtained with a spinning-disc confocal scanner. Cerebellar slices were from transgenic mouse lines (L7-tau-GFP shown; pcp2-EGFP was also used) in which GFP or EGFP was expressed in Purkinje cells. B, Voltage-sensitive dye fluorescence image of a part of the axonal arbor in recording position obtained with the CCD camera for voltage imaging (different cell from A).