Cells respond to external biochemical and mechanical cues to generate myriad responses including force generation, motility, gene expression and differentiation. The connection between external cue and internal response are the biochemical pathways that control the cell. Understanding these responses is critical to gaining insight into diverse biomedical phenomena such as cancer, immunity and tissue differentiation and morphogenesis. Our workshop will offer, through morning lectures and afternoon laboratory sessions, ways to assess the biochemistry and mechanobiology of cells, and the theoretical basis for interpreting physical properties and force sensing and generation. We will cover the use of FRET-based biosensors related to cell mechanics and responses, such as RhoA and Rac1, plus light-modulated signaling molecules. We will present the optics and plans for assembling your own versatile light control microscopy systems that includes vertical light sheet microscopy, spinning total internal reflection fluorescence (TIRF) and photoactivation capability. New to this year’s workshop will be two morning sessions on image analysis using the FIJI toolbox, taught by guest speaker Teng-Leong Chew from HHMI Janelia Research Center. Experimental stations will include atomic force microscopy combined with light sheet microscopy, 3D force microscopy (magnetic tweezers) with TIRF, the use of a high throughput, 12 channel microscope for cell rheology and FRET microscopy using fluorescent biosensors.