The first application of these biosensors revealed that talin, a molecule that connects integrin receptors with the actin cytoskeleton through multiple interaction sites, indeed experiences mechanical forces of about 7–10 pN. A particularly interesting finding of this study was that binding of talin’s interaction partner vinculin is crucial to generate high tension across talin. Importantly, this vinculin-dependent increase in talin tension could only be resolved with HP35- but not with TSMod-based talin sensors (Austen et al., 2015). We therefore suggest that future experiments use all three currently available single-molecule calibrated tension sensors in parallel to more accurately determine mechanical forces across distinct proteins in cells (Fig. 3). However, such experiments will be still limited by the tension threshold of HP35st and forces above 10–12 pN will remain unresolved.