  Live cell imaging of focal adhesions requires a sufficiently high temporal
resolution, which remains a challenging task for super-resolution microscopy.
We have addressed this important issue by combining photo-activated
localization microscopy (PALM) with super-resolution optical fluctuation
imaging (SOFI). Using simulations and fixed cell focal adhesion images, we
investigated the complementarity between PALM and SOFI in terms of spatial and
temporal resolution. This PALM-SOFI framework was used to image focal adhesions
in living cells, while obtaining a temporal resolution below 10 s. We
visualized the dynamics of focal adhesions, and revealed local mean velocities
around 190 nm per minute. The complementarity of PALM and SOFI was assessed in
detail with a methodology that integrates a quantitative resolution and
signal-to-noise metric. This PALM and SOFI concept provides an enlarged
quantitative imaging framework, allowing unprecedented functional exploration
of focal adhesions through the estimation of molecular parameters such as the
fluorophore density and the photo-activation and photo-switching rates.
