Axial displacement calibration and tracking of optically trapped beads

Abstract

High-precision axial displacement tracking of trapped beads is an indispensable feature of optical tweezers in advanced single-molecule studies. Here, we demonstrate an alternative method that enables axial calibration and tracking to be carried out on the same sample to avoid unnecessary errors. This method works by applying a dynamic force balance on a bead trapped between a piezoelectrically driven glass slide and an optical trap; in this configuration, the bead can be stopped precisely in different positions and imaged by a camera. A simple gradient algorithm is used to process the images into calibration data. After optimization of the calibration method and samples, our method exhibited better than 5 nm experimental axial resolution, with a measurement range of +/-500 nm around the objective focus at video speed. Moreover, for the first time, the deviation of the focusing plane in dual-trap optical tweezers was measured. We confirmed the axial deviation between two optical traps in our setup to be ~10 nm, corresponding to a force spectroscopy gage error of ~1 pN. This approach offers a favorable solution for in-use setup updating, as it can be seamlessly integrated into any optical tweezers system without requiring new hardware updates.