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Tapping mode atomic force microscopy (AFM) was used to investigate the topography of isolated ryanodine receptors of rabbit skeletal muscle (RyR1) both in air and in a buffer. In air, images fourfold symmetric in appearance were obtained. Two different configurations could be distinguished in the AFM topography of single RyR1s, depending on whether there was a center protrusion. In the buffer, even though square images of RyR1s were discerned, the detailed topography of RyR1 appeared different from that in air. Possible reasons for this are discussed. © 2000 American Vacuum Society.