Measurement of diffusion parameters using a sinusoidal iontophoretic source in rat cortex.

A new method was developed to extract diffusion parameters in brain tissue using a sinusoidal iontophoretic point source of tetramethylammonium operated at different frequencies. The resulting steady state oscillating extracellular concentration of this probe molecule was continuously monitored using an ion-selective microelectrode located about 100 microm from the source. Because the probe molecules must diffuse through the extracellular space (ECS), the oscillating concentration at the recording location will develop a phase lag and an amplitude attenuation relative to the sinusoidal source. These two components of the signal can be analyzed to determine the tortuosity factor lambda and the ECS volume fraction alpha. The method also measures the nonspecific clearance rate constant kappa. In brain slices this reflects washout of diffusing molecules. Values of alpha (0.18+/-0.05) and lambda (1.67+/-0.08) obtained from this frequency method in rat cortical slices were similar to those obtained by the real-time iontophoretic method employing a square pulse source. The relative merits of the frequency method compared to the pulse method are discussed.