  Aiming at absolute force calibration of optical tweezers, following a
critical review of proposed theoretical models, we present and test the results
of MDSA (Mie-Debye-Spherical Aberration) theory, an extension of a previous
(MD) model, taking account of spherical aberration at the glass/water
interface. This first-principles theory is formulated entirely in terms of
experimentally accessible parameters (none adjustable). Careful experimental
tests of the MDSA theory, undertaken at two laboratories, with very different
setups, are described. A detailed description is given of the procedures
employed to measure laser beam waist, local beam power at the transparent
microspheres trapped by the tweezers, microsphere radius and the trap
transverse stiffness, as a function of radius and height in the (inverted
microscope) sample chamber. We find generally very good agreement with MDSA
theory predictions, for a wide size range, from the Rayleigh domain to large
radii, including the values most often employed in practice, and at different
chamber heights, both with objective overfilling and underfilling. The results
asymptotically approach geometrical optics in the mean over size intervals, as
they should, and this already happens for size parameters not much larger than
unity. MDSA predictions for the trapping threshold, position of stiffness peak,
stiffness variation with height, multiple equilibrium points and `hopping'
effects among them are verified. Remaining discrepancies are ascribed to focus
degradation, possibly arising from objective aberrations in the infrared, not
yet included in MDSA theory.
