Figure 1.

Flow-chart of a typical experiment. Cells were mounted in the perfusion chamber and equilibrated in PS (37°C, ∼10 min) (O); perfusion with ILS (1); permeabilization with digitonin (2 min) or amphotericin B (8 min) followed by the wash out (2); change of osmolality, pH, or [Mg²⁺] of the external solution (3); return to initial conditions to test reversibility of the volume changes (4). Trypan Blue added at this point to verify that the cell remained permanently permeabilized.