Optical trapping of sub-millimeter sized particles and microorganisms.

While optical tweezers (OT) are mostly used for confining smaller size particles, the counter-propagating (CP) dual-beam traps have been a versatile method for confining both small and larger size particles including biological specimen. However, CP traps are complex sensitive systems, requiring tedious alignment to achieve perfect symmetry with rather low trapping stiffness values compared to OT. Moreover, due to their relatively weak forces, CP traps are limited in the size of particles they can confine which is about 100 μm. In this paper, a new class of counter-propagating optical tweezers with a broken symmetry is discussed and experimentally demonstrated to trap and manipulate larger than 100 μm particles inside liquid media. Our technique exploits a single Gaussian beam folding back on itself in an asymmetrical fashion forming a CP trap capable of confining small and significantly larger particles (up to 250 μm in diameter) based on optical forces only. Such optical trapping of large-size specimen to the best of our knowledge has not been demonstrated before. The broken symmetry of the trap combined with the retro-reflection of the beam has not only significantly simplified the alignment of the system, but also made it robust to slight misalignments and enhances the trapping stiffness as shown later. Moreover, our proposed trapping method is quite versatile as it allows for trapping and translating of a wide variety of particle sizes and shapes, ranging from one micron up to a few hundred of microns including microorganisms, using very low laser powers and numerical aperture optics. This in turn, permits the integration of a wide range of spectroscopy techniques for imaging and studying the optically trapped specimen. As an example, we will demonstrate how this novel technique enables simultaneous 3D trapping and light-sheet microscopy of C. elegans worms with up to 450 µm length.