The pathways and mechanisms by which edematous fluid accumulation in the extracellular space (ECS) clears from brain are poorly understood. The objective of this study was to explore, using immunocytochemical technique, the fate of a proteinaceous fluid added to the brain ECS and to study the clearance pathways. The protein movement of this edema fluid was investigated using the direct infusion model on rats. Rat albumin (20 microliters) was slowly infused into the caudate-putamen of anesthetized adult rats and the spread and clearance of the edema was followed in various brain regions using immunocytochemical and conventional light and electron microscopy at 0, 1, 2, 3, 4, 6, and 8 days post-infusion. Our studies showed that protein-rich edema fluid cleared slowly from the brain, with 8 days required for the infusion albumin to exit completely from the brain parenchyma. Immediately following infusion, the albumin was distributed in the ECS of the white matter and the overlying deep cortical layers related to the infusion site. During the next 24 h, more of the infused albumin traveled through the ECS to the cortical surface where the albumin passed through the glia limitans to reach the subarachnoid front. Additionally, at 48 h post-infusion, that albumin, which had migrated to the ventricular wall, cleared from the ECS of the subependymal white matter and the ependymal clefts to reach the ventricular cerebrospinal fluid (CSF). In edematous regions, the perivascular spaces of venules and veins were filled with reaction product. Continuity of this perivascular reaction product existed from the deep edematous area to the temporobasal subarachnoid space from where the reaction product gradually disappeared from the parenchyma. From these studies we infer that during the late state of the resolution process the edema front moves toward both the ventricle and the cortical surface to reach the CSF. Thus, among the potential routes for edema clearance, the pathways leading to CSF clearance of fluid predominated. During this clearance process, neither neurons, glia nor the vascular endothelium showed any endocytotic response to the infused albumin throughout the 8-day course. We conclude from these observations that the CSF pathway is the major route of protein-rich edema clearance, when such clearance is not complicated by any concomitant CNS perturbation.