Independent and simultaneous three-dimensional optical trapping and imaging

Combining imaging and control of multiple micron-scaled objects in three dimensions opens up new experimental possibilities such as the fabrication of colloidal-based photonic devices, as well as high-throughput studies of single cell dynamics. Here we utilize the dual-objectives approach to combine 3D holographic optical tweezers with a spinning-disk confocal microscope. Our setup is capable of trapping multiple different objects in three dimensions with lateral and axial accuracy of 8 nm and 20 nm, and precision of 20 nm and 200 nm respectively, while imaging them in four different fluorescence channels. We demonstrate fabrication of ordered two-component and three dimensional colloidal arrays, as well as trapping of yeast cell arrays. We study the kinetics of the division of yeast cells within optical traps, and find that the timescale for division is not affected by trapping.