Maximizing precision in quantitative autoradiographic determination of tissue tracer concentration using exposure optimization.

The relationships between tissue tracer concentrations, length of time of tissue contact with film, and the darkness of resulting autoradiographic images were explored. Operational equations were then developed relating ranges of film darkening to the ranges of tracer concentration contained within the sections. These equations were solved and used to define ranges of optical density that are optimal for precise determination of tracer concentration using digital image analyzers. The solutions indicated that optimal optical densities are a function of the range of tracer concentrations in the sections. For autoradiograms of typical cerebral blood flow and metabolism tracers, exposure should be controlled to produce images that are significantly less dark than what is generally considered pleasing to the eye.