Summary High resolution atomic force microscopy (AFM) can be per- formed simultaneously with optical microscopy techniques, such as fluorescence or differential interference contrast (DIC) microscopy. The combined methodologies provide comple- mentary information about the studied sample which establishes the basis for a better understand- ing of physiological processes and the function of biomolecules, and allow a more detailed characteriza- tion of the structure of living cells [1]. Here we present the Agilent 5500 ILM AFM/SPM, which is an easy-to-use AFM and inverted light microscope (ILM) combination that enhances live-cell imaging under physiological conditions. The advantage of this combined setup is further demonstrated by studying fluorescence-labeled membrane receptors via force spectroscopy using antibody- functionalized AFM tips.