ABSTRACT

This chapter presents an experimental strategy based on atomic force microscope (AFM) force sensing on individual cells to report on the effects of external pharmacological stimuli on cellular functions. It shows that AFM-based force measurements in conjunction with fluorescence imaging of intracellular component can fingerprint the contribution of signalling pathways subsequent to the activation of specific receptor at the cell membrane. Compared with the data obtained using cell-free assays, direct monitoring of drug-modulated signalling in live cell systems offers high content information, in conditions designed to closely resemble a physiological environment. To delineate the contribution of the signalling pathways activated by a given receptor in the development of mechanical response in individual cells, one should decouple or isolate their contributions. Stimulation of cell receptors leads to the activation of several distinct signalling pathways involving a variety of structural and functional intracellular proteins and biochemical second messengers.