The effect of varying perfusion medium osmolality on the in vivo microdialysis recovery of caffeine and 5-chloro-2',3'dideoxy-3'-fluorouridine (FCdU) was investigated. The microdialysis probes were implanted in rats into the corpus striatum of the brain. Each probe was perfused with a solution containing 10 mu M FCdU (in distilled water, Ringer's solution or in 20% sucrose in Ringer's solution) or with a solution containing 10 mu M caffeine (in distilled water, Ringer's solution, in 20% sucrose in distilled water or in 20% sucrose in Ringer's solution). Recovery was calculated as the proportion of concentration loss over the microdialysis membrane to the perfusion medium (i.e. from 10 mu M). The recovery was significantly dependent on the composition of the perfusion medium. The recoveries of FCdU were 0.097 (55%) for water solution, 0.176 (100%) for Ringer's solution and 0.292 (166%) for sucrose's solution. The recoveries of caffeine were 0.196 (73%) for water solution, 0.270 (100%) for Ringer's solution and 0.335 (124%) for sucrose's solution in water and 0.303 (112%) for sucrose's solution in Ringer. A linear correlation between the recovery over microdialysis membrane and osmolality of perfusion medium was demonstrated for FCdU. The model describing the change of the recovery of caffeine with osmolality of perfusion medium fitted a polynomial function of second order. It is concluded that the osmolality of the perfusion medium influences the recovery in microdialysis experiments. A possible mechanism for this phenomenon is an increase in the extracellular volume fraction with increasing osmolality, thereby increasing the effective diffusion in the tissue. (C) 1999 Elsevier Science B.V. All rights reserved.