Eukaryotic cells use membrane organelles, like the endoplasmic reticulum or the Golgi, to carry out different functions. Vertebrate rod photoreceptors use hundreds of membrane sacs (the disks) for the detection of light. We have used fluorescent tracers and single cell imaging to study the properties of rod photoreceptor disks. Labeling of intact rod photoreceptors with membrane markers and polar tracers revealed communication between intradiskal and extracellular space. Internalized tracers moved along the length of the rod outer segment, indicating communication between the disks as well. This communication involved the exchange of both membrane and aqueous phase and had a time constant in the order of minutes. The communication pathway uses approximately 2% of the available membrane disk area and does not allow the passage of molecules larger than 10 kDa. It was possible to load the intradiskal space with fluorescent Ca(2+) and pH dyes, which reported an intradiskal Ca(2+) concentration in the order of 1 microM and an acidic pH 6.5, both of them significantly different than intracellular and extracellular Ca(2+) concentrations and pH. The results suggest that the rod photoreceptor disks are not discrete, passive sacs but rather comprise an active cellular organelle. The communication between disks may be important for membrane remodeling as well as for providing access to the intradiskal space of the whole outer segment.