Abstract
In this report, we combine total internal reflection-fluorescence correlation spectroscopy (TIR-FCS) with a single optical
trap to simultaneously manipulate and measure the dynamics of individual molecules near the substrate-solution
interface. As a proof of principle, polystyrene particles (84 nm in diameter) are used as a model system to test our
approach in studying their diffusion properties near surfaces, which are treated with polyethylene glycol 8000, bovine
serum albumin or sodium hydroxide. The evanescent field of 543 nm excitation propagates ~100 nm into the solution,
and the fluorescence detection is spatially confined by a 25 or 50 μm pinhole that is parfocal with the specimen plane.
The optical trap is generated using a cw Ti:sapphire laser at 780 nm. Our results indicate that the particles' diffusion is
influenced by surface interactions, which might have further implications on biomembrane studies. Furthermore, the
observed translational diffusion of individual particles can be manipulated using an optical trap. By combining the single
molecule sensitivity of TIR-FCS with a noninvasive manipulation method, such as optical trapping, we will be able to
probe molecular dynamics in biomimetic systems and living cells.
