Intercellular diffusion of macromolecules is observed between fibers located deep within the lens but not between superficial cells. (A) Injections of 10 kDa TMRD (red) were made into individual fiber cells situated at different depths below the surface of a living P2 lens. The 0 hour time point is a nominal value. It took approximately 10 minutes to complete the injections and photograph the lens. During the interim TMRD had already begun to diffuse from injected cells in the lens core (arrowheads). (B) TMRD injected into surface cells (blue arrows) was well retained, even 12 hours after the injection. By contrast, TRMD was not retained by injected cells in the center of the lens (arrowheads) and, after 12 hours, formed a diffuse cloud of fluorescence in the lens core (*). The results are representative of 12 experiments. Bar, 250 μm.