single-molecule force spectroscopy measures force and extension of an individual tether while controlling position or load. In an atomic-force arrangement, a synthetic chain connects a cantilever probe to a translated substrate. For the cantilever compliance correction, a deflection sensor gives , a calibrated spring constant gives , and substrate displacement gives chain extension after geometric offsets are removed. Additional linkers and instrument compliance must be included if appreciable. Thermal deflection calibration uses only after isolating the cantilever's calibrated thermal contribution.
An optical-tweezers implementation detects a tethered bead's displacement from a calibrated trap centre, with in the harmonic regime. Detector calibration, drag and trap linearity determine the usable force and frequency range. These instruments measure individual force–extension curves and distributions; averaging many curves can hide multiple tether populations.
For a synthetic freely jointed chain with Kuhn length ,A strongly stretched worm-like chain instead has before backbone stretching becomes important. Fits can estimate contour and persistence lengths only in their appropriate regimes. Hysteresis and pulling-rate dependence test whether an apparent mechanical transition is quasistatic or activated.
Control single versus multiple attachments, subtract nonspecific adhesion, vary pulling speed, calibrate temperature and stiffness, and correct drift. A rupture peak is not a universal bond strength independent of loading history. A primary nonbiological example measured individual polydimethylsiloxane chains by atomic-force methods; its flexibility interpretation combines those measurements with molecular calculations.[ref]
