Implementing both short- and long-working-distance optical trappings into a commercial microscope

Abstract

Optical tweezers are now a widespread tool based on three-dimensional trapping by a tightly focused single laser beam. This configuration only works with large numerical aperture and short-working-distance (SWD) objectives, restricting optical manipulation to the high magnification end of the microscope nosepiece. Certain applications of optical trapping demand long-working distances (LWDs) at moderate magnification, imposing a more complex two-beam trapping configuration. In this article, we describe a complete setup that incorporates both SWD and LWD optical trapping functionalities into a single Axiovert 200M Zeiss microscope. We evaluate the performance of the setup in both trapping modes with latex particles, either fluorescent or not, of different sizes, in the 1–20μm range. We provide practical information allowing for optimal configuration of the two-beam geometry, in relation with longitudinal and lateral stabilities of the trap.