The most basic optical tweezer setup will likely include the following components: a laser (usually
Nd:YAG), a beam expander, some optics used to steer the beam location in the sample plane, a
microscope objective and
condenser to create the trap in the sample plane, a position detector (e.g. quadrant
photodiode) to measure beam displacements and a microscope illumination source coupled to a
CCD camera (typically via an
optical mount. An
Nd:YAG laser (1064 nm wavelength) is a common choice of laser for working with biological specimens. This is because such specimens (being mostly water) have a low
absorption coefficient at this wavelength. A low absorption is advisable so as to minimise damage to the biological material, sometimes referred to as
opticution. Perhaps the most important consideration in optical tweezer design is the choice of the objective. A stable trap requires that the gradient force, which is dependent upon the
numerical aperture (NA) of the objective, be greater than the scattering force. Suitable objectives typically have an NA between 1.2 and 1.4. While alternatives are available, perhaps the simplest method for position detection involves imaging the trapping laser exiting the sample chamber onto a quadrant photodiode. Lateral deflections of the beam are measured similarly to how it is done using
atomic force microscopy (AFM). Expanding the beam emitted from the laser to fill the
aperture of the objective will result in a tighter, diffraction-limited spot. While lateral translation of the trap relative to the sample can be accomplished by translation of the microscope slide, most tweezer setups have additional optics designed to translate the beam to give an extra degree of translational freedom. This can be done by translating the first of the two lenses labelled as "Beam Steering" in the figure. For example, translation of that lens in the lateral plane will result in a laterally deflected beam from what is drawn in the figure. If the distance between the beam steering lenses and the objective is chosen properly, this will correspond to a similar deflection before entering the objective and a resulting
lateral translation in the sample plane. The position of the beam waist, that is the focus of the optical trap, can be adjusted by an axial displacement of the initial lens. Such an axial displacement causes the beam to diverge or converge slightly, the result of which is an axially displaced position of the beam waist in the sample chamber. Visualization of the sample plane is usually accomplished through illumination via a separate light source coupled into the optical path in the opposite direction using
dichroic mirrors. This light is incident on a CCD camera and can be viewed on an external monitor or used for tracking the trapped particle position via
video tracking.
Alternative laser beam modes The majority of optical tweezers make use of
conventional TEM00 Gaussian beams. However a number of other beam types have been used to trap particles, including high order laser beams i.e.
Hermite-Gaussian beams (TEMxy),
Laguerre-Gaussian (LG) beams (TEMpl) and
Bessel beams. Optical tweezers based on Laguerre-Gaussian beams have the unique capability of trapping particles that are optically reflective and absorptive. Laguerre-Gaussian beams also possess a well-defined
orbital angular momentum that can rotate particles. This is accomplished without external mechanical or electrical steering of the beam. Both zero and higher order Bessel Beams also possess a unique tweezing ability. They can trap and rotate multiple particles that are millimeters apart and even around obstacles.
Micromachines can be driven by these unique optical beams due to their intrinsic rotating mechanism due to the
spin and orbital angular momentum of light.
Multiplexed optical tweezers A typical setup uses one laser to create one or two traps. Commonly, two traps are generated by splitting the laser beam into two orthogonally polarized beams. Optical tweezing operations with more than two traps can be realized either by time-sharing a single laser beam among several optical tweezers, or by diffractively splitting the beam into multiple traps. With acousto-optic deflectors or
galvanometer-driven mirrors, a single laser beam can be shared among hundreds of optical tweezers in the focal plane, or else spread into an extended one-dimensional trap. Specially designed diffractive optical elements can divide a single input beam into hundreds of continuously illuminated traps in arbitrary three-dimensional configurations. The trap-forming hologram also can specify the mode structure of each trap individually, thereby creating arrays of optical vortices, optical tweezers, and holographic line traps, for example. When implemented with a
spatial light modulator, such holographic optical traps also can move objects in three dimensions. Advanced forms of holographic optical traps with arbitrary spatial profiles, where smoothness of the intensity and the phase are controlled, find applications in many areas of science, from micromanipulation to
ultracold atoms. Ultracold atoms could also be used for realization of quantum computers.
Single mode optical fibers The standard fiber optical trap relies on the same principle as the optical trapping, but with the Gaussian laser beam delivered through an
optical fiber. If one end of the optical fiber is molded into a
lens-like facet, the nearly gaussian beam carried by a single mode standard fiber will be focused at some distance from the fiber tip. The effective Numerical Aperture of such assembly is usually not enough to allow for a full 3D optical trap but only for a 2D trap (optical trapping and manipulation of objects will be possible only when, e.g., they are in contact with a surface ). A true 3D optical trapping based on a single fiber, with a trapping point which is not in nearly contact with the fiber tip, has been realized based on a not-standard annular-core fiber arrangement and a total-internal-reflection geometry. On the other hand, if the ends of the fiber are not moulded, the laser exiting the fiber will be diverging and thus a stable optical trap can only be realised by balancing the gradient and the scattering force from two opposing ends of the fiber. The gradient force will trap the particles in the transverse direction, while the
axial optical force comes from the scattering force of the two counter propagating beams emerging from the two fibers. The equilibrium z-position of such a trapped bead is where the two scattering forces equal each other. This work was pioneered by A. Constable
et al.,
Opt. Lett. 18,1867 (1993), and followed by J.Guck
et al.,
Phys. Rev. Lett. 84, 5451 (2000), who made use of this technique to stretch microparticles. By manipulating the input power into the two ends of the fiber, there will be an increase of an "optical stretching" that can be used to measure viscoelastic properties of cells, with sensitivity sufficient to distinguish between different individual cytoskeletal phenotypes. i.e. human erythrocytes and mouse fibroblasts.
Multimode fiber-based traps "Optical cell rotator" (OCR) technology decouples trapping from imaging optics. This, its modular design, and the high compatibility of divergent laser traps with biological material indicates the great potential of this new generation of laser traps in medical research and life science. Based on
adaptive optics, OCR allow the dynamic reconfiguring the optical trap during operation and adapt it to the sample.
Cell sorting One of the more common cell-sorting systems makes use of flow cytometry through
fluorescence imaging. In this method, a suspension of biologic cells is sorted into two or more containers, based upon specific fluorescent characteristics of each cell during an assisted flow. By using an electrical charge that the cell is "trapped" in, the cells are then sorted based on the fluorescence intensity measurements. The sorting process is undertaken by an electrostatic deflection system that diverts cells into containers based upon their charge. In the optically actuated sorting process, the cells are flowed through into an optical landscape i.e. 2D or 3D optical lattices. Without any induced electrical charge, the cells would sort based on their intrinsic refractive index properties and can be re-configurability for dynamic sorting. An optical lattice can be created using diffractive optics and optical elements. K. Xiao and D. G. Grier applied holographic video microscopy to demonstrate that this technique can sort colloidal spheres with part-per-thousand resolution for size and refractive index. The main mechanism for sorting is the arrangement of the optical lattice points. As the cell flow through the optical lattice, there are forces due to the particles
drag force that is competing directly with the optical gradient force
(See Physics of optical tweezers) from the optical lattice point. By shifting the arrangement of the optical lattice point, there is a preferred optical path where the optical forces are dominant and biased. With the aid of the flow of the cells, there is a resultant force that is directed along that preferred optical path. Hence, there is a relationship of the flow rate with the optical gradient force. By adjusting the two forces, one will be able to obtain a good optical sorting efficiency. Competition of the forces in the sorting environment need fine tuning to succeed in high efficient optical sorting. The need is mainly with regards to the balance of the forces; drag force due to fluid flow and optical gradient force due to arrangement of intensity spot. Scientists at the University of St. Andrews have received considerable funding from the UK
Engineering and Physical Sciences Research Council (
EPSRC) for an optical sorting machine. This new technology could rival the conventional fluorescence-activated cell sorting.
Evanescent fields An
evanescent field is a residue
optical field that "leaks" during
total internal reflection. This "leaking" of light fades off at an exponential rate. The evanescent field has found a number of applications in nanometer resolution imaging (microscopy); optical micromanipulation (optical tweezers) are becoming ever more relevant in research. In optical tweezers, a continuous evanescent field can be created when light is propagating through an
optical waveguide (multiple
total internal reflection). The resulting evanescent field has a directional sense and will propel microparticles along its propagating path. This work was first pioneered by S. Kawata and T. Sugiura, in 1992, who showed that the field can be coupled to the particles in proximity on the order of 100 nanometers. This direct coupling of the field is treated as a type of photon tunnelling across the gap from prism to microparticles. The result is a directional optical propelling force. The evanescent field generated by mid-infrared laser has been used to sort particles by molecular vibrational resonance selectively. Mid-infrared light is commonly used to identify molecular structures of materials because the vibrational modes exist in the mid-infrared region. Optical force enhancement by molecular vibrational resonance can be achieved by exciting the stretching mode of Si-O-Si bond at 9.3 μm. It is shown that silica microspheres containing significant Si-O-Si bond move up to ten times faster than polystyrene microspheres due to molecular vibrational resonance. Moreover, this same group also investigated the possibility of optical force chromatography based on molecular vibrational resonance. Surface plasmons, an enhanced evanescent wave localized at a metal/dielectric interface has been demonstrated using a photonic force microscope, the total force magnitude being found 40 times stronger compared to a normal evanescent wave. By patterning the surface with gold microscopic islands it is possible to have selective and parallel trapping in these islands. The forces of the latter optical tweezers lie in the femtonewton range. The evanescent field can also be used to trap
cold atoms and molecules near the surface of an optical waveguide or
optical nanofiber.
Indirect approach Ming Wu, a
UC Berkeley Professor of electrical engineering and computer sciences invented the new optoelectronic tweezers. Wu transformed the optical energy from low powered light emitting diodes (LED) into electrical energy via a photoconductive surface. The idea is to allow the LED to switch on and off the photoconductive material via its fine projection. As the optical pattern can be easily transformable through optical projection, this method allows a high flexibility of switching different optical landscapes. The manipulation/tweezing process is done by the variations between the electric field actuated by the light pattern. The particles will be either attracted or repelled from the actuated point due to its induced electrical dipole. Particles suspended in a liquid will be susceptible to the electrical field gradient, this is known as
dielectrophoresis. One clear advantage is that the electrical conductivity is different between different kinds of cells. Living cells have a lower conductive medium while the dead ones have minimum or no conductive medium. The system may be able to manipulate roughly 10,000 cells or particles at the same time. See comments by Professor Kishan Dholakia on this new technique, K. Dholakia,
Nature Materials 4, 579–580 (01 Aug 2005) News and Views. "The system was able to move live E. coli bacteria and 20-micrometre-wide particles, using an optical power output of less than 10 microwatts. This is one-hundred-thousandth of the power needed for [direct] optical tweezers". Another notably new type of optical tweezers is optothermal tweezers invented by Yuebing Zheng at
The University of Texas at Austin. The strategy is to use light to create a temperature gradient and exploit the thermophoretic migration of matter for optical trapping. The team further integrated thermophoresis with
laser cooling to develop opto-refrigerative tweezers to avoid thermal damages for noninvasive optical trapping and manipulation.
Optical binding When a cluster of microparticles are trapped within a monochromatic laser beam, the organization of the microparticles within the optical trapping is heavily dependent on the redistributing of the optical trapping forces amongst the microparticles. This redistribution of light forces amongst the cluster of microparticles provides a new force equilibrium on the cluster as a whole. As such we can say that the cluster of microparticles are somewhat bound together by light. One of the first experimental evidence of optical binding was reported by Michael M. Burns, Jean-Marc Fournier, and Jene A. Golovchenko, though it was originally predicted by T. Thirunamachandran.
Fluorescence optical tweezers In order to simultaneously manipulate and image samples that exhibit
fluorescence, optical tweezers can be built alongside a
fluorescence microscope. Such instruments are particularly useful when it comes to studying single or small numbers of biological molecules that have been fluorescently labelled, or in applications in which fluorescence is used to track and visualize objects that are to be trapped. This approach has been extended for simultaneous sensing and imaging of dynamic protein complexes using long and strong tethers generated by a highly efficient multi-step enzymatic approach{{cite journal |vauthors= Avellaneda MJ, Koers EJ, Minde DP, Sunderlikova V, Tans SJ |title= Simultaneous sensing and imaging of individual biomolecular complexes enabled by modular DNA–protein coupling|journal=Communications Chemistry |volume=3 |pages=1–7 |year=2020 |issue= 1|article-number= 20|doi=10.1038/s42004-020-0267-4 == See also ==