Microscopic observations on endogenous fluorochromes within a nerve fibre excited by a 325 nm He-Cd laser

A microscopic procedure to observe endogenous fluorochromes in superfused tissues is described. In this procedure, a 325 nm beam from a He-Cd laser is directed through the objective lens of a microscope onto the surface of a brain slice. Fluorescent structures within the tissue can be observed, photographed and measured. A nerve fibre running from the corporis callosi to the dorsal cerebral cortex was studied with this approach. The 325 nm laser beam excited material with an emission peak at 420 nm within a fibre on the cut surface of a rat brain slice superfused with Krebs-Ringer. A similar fibre was observed in vivo in the exposed mouse brain. Packets of the fluorescent material appear to move within the fibre in vitro. The fluorescence of the fibre was rapidly diminished by cyanide, colchicine, rotenone and the withdrawal of Ca2+. Elevation of [K+] in the superfusate caused a "haloing' of the fibre's granularity without decreasing the total fluorescence. This phenomenon is possibly related to the depolarization by [K+] of neurones. Although the structure of the fluorochrome was not established, it is suggested that it may be a neurotransmitter-related substance undergoing axonal movement.