We investigated the adsorption of albumin on chemically modified gold surfaces by scanning force microscopy operating both in contact and noncontact mode. The surface modification was performed with thiol‐based selfassembling molecules carrying amine or methyl groups. The albumin on the aminoethanethiol‐coated gold formed a uniform layer and single molecules could be distinguished. On the dodecanethiol‐coated surface the protein adsorbed in aggregates or single isolated molecules depending on the incubation time. The width of the albumin molecule on both surfaces was similar, but the height was much lower on the amine than on the methyl surface. This was interpreted as a difference in the conformation of albumin depending on the substrate, and could explain the promotion of cell adhesion on amine‐treated polymers coated with albumin.