Action potential-induced fluorescence changes resolved with an optical fiber carrying excitation light

With the use of a single, implantable, optical fiber, to excite fluorescence and detect changes from voltage-sensitive dyes, transmembrane potential changes were measured without the need for a clear line-of-sight path between the excitation light, the tissue, and the detector. In a previous study, we were required to use signal averaging and could detect only cardiac action potentials from frog. In the present study we improved this system so that unaveraged cardiac action potentials were resolved with high fidelity, and action potentials from single nerve axons were detected. Endeavors to optimize the signal-to-noise ratio resulted in the selection of a larger core fiber with a rounded tip, styryl dyes, and filters based upon fluorescence spectra of the dyes when bound to membrane (rather than in solution). The frog gave signals nearly comparable in magnitude and signal-to-noise ratio to those seen with systems that use a fluorescence microscope. Action potential-induced signals could be detected in single lobster axons with the intracellular injection of a dye. The improvement in the signal-to-noise ratio allowed the use of a reduced-intensity excitation illumination which produced less bleaching of the dye.