Visualization of calcium-binding sites at plasma membrane of shock-frozen Acanthamoeba cells

Electron-dense deposits appear at the protoplasmic side of plasma membrane in Acanthamoeba log-phase cells when fixed either with glutaraldehyde or formaldehyde supplemented with Ca2+. Similar deposits appear when the cells are preloaded with Ca2+ and thereafter shock-frozen and prepared for electron microscopic examination by freeze-substitution technique. This suggests that their formation reflects the presence of a physiologically active system involved in capturing excess of inflowing calcium.