A new type of flat substrate has been used to visualize structures inside living cells by surface-enhanced Raman scattering (SERS) and to study biochemical processes within cells. The SERS substrate is formed by stabilized aggregates of gold nanostars on a glass microscope slide coated with a layer of poly (4-vinyl pyridine) polymer. This type of SERS substrate provides good cell adhesion and viability. Au nanostars’ long tips can penetrate the cell membrane, allowing it to receive the SERS signal from biomolecules inside a living cell. The proposed nanostructured surfaces were tested to study, label-free, the distribution of various biomolecules in cell compartments.