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Oblique illumination imaging can significantly improve the contrast of transparent thin samples. However, in traditional oblique illumination methods, either the condenser is offset or a block is added to the condenser, which makes it complicated and challenged to build a stable oblique illumination imaging. Herein, we present a method to measure the optimal shading ratio of oblique illumination in an inverted microscope, and develop an apparatus for stable high-speed high-contrast imaging with uniform brightness. At optimal shading ratio, the oblique illumination imaging has better imaging quality than differential interference contrast, which characteristic is independent on sample. In oblique illumination with low magnification objective, the images have uneven brightness. According to target brightness, we have developed a brightness unevenness correction algorithm to form uniform background brightness for oblique illumination. Integrating the algorithm with imaging acquisition, corrected oblique illumination microscopy is appropriate to observe living cells with high contrast.

Cell assembly has important applications in biomedical research, which can be achieved with laser-heating induced thermal convective flow. In this paper, an opto-thermal approach is developed to assemble the yeast cells dispersed in solution. At first, polystyrene (PS) microbeads are used instead of cells to explore the method of microparticle assembly. The PS microbeads and light absorbing particles (APs) are dispersed in solution and form a binary mixture system. Optical tweezers are used to trap an AP at the substrate glass of the sample cell. Due to the optothermal effect, the trapped AP is heated and a thermal gradient is generated, which induces a thermal convective flow. The convective flow drives the microbeads moving toward and assembling around the trapped AP. Then, the method is used to assemble the yeast cells. The results show that the initial concentration ratio of yeast cells to APs affects the eventual assembly pattern. The binary microparticles with different initial concentration ratios assemble into aggregates with different area ratios. The experiment and simulation results show that the dominant factor in the area ratio of yeast cells in the binary aggregate is the velocity ratio of the yeast cells to the APs. Our work provides an approach to assemble the cells, which has a potential application in the analysis of microbes.

The photothermal effects have shown the possibilities for applications in optical manipulation. In this paper, an approach is demonstrated to generate and manipulate a bubble using the photothermal effects. First, a high-power laser is used to irradiate the light absorbing particles for creating a microbubble. The bubble grows up to a diameter of a few hundred micrometers in several seconds due to the diffusion of dissolved gases. The bubble does not float up and is confined at the lower boundary of the sample cell by the thermocapillary force. The force is induced by laser heating of the particles at the bubble base. Second, the bubble can be manipulated following the laser focal spot. The bubble is dragged by the horizontal component of thermocapillary force. The bubble re-grows as it moves because it absorbs the dissolved gases in its migration path. The bubble floats up finally when it grows up to the maximum size. The perpendicular component of thermocapillary force can be estimated equal to the buoyancy of the floated bubble and is about 38 nN at the laser power of 130 mW. Furthermore, we show the generation and manipulation of the bubbles in a capillary. The reason for the decrease in movement velocity in the capillaries has been studied and discussed. The approach of bubble manipulation shows a potential application in transporting the microparticles.

The ability to trap and rotate magnetic particles has important applications in biophysical research and optical micromachines. However, it is difficult to achieve the spin rotation of magnetic particles with optical tweezers due to the limit in transferring spin angular momentum of light. Here, we propose a method to obtain controlled spin rotation of a magnetic microparticle by the phoretic torque, which is originated from inhomogeneous heating of the microparticle’s surface. The microparticle is trapped and rotated nearby the laser focus center. The rotation frequency is several Hertz and can be controlled by adjusting the laser power. Our work provides a method to the study of optical rotation of microscopic magnetic particles, which will push toward both translational and rotational manipulation of the microparticles simultaneously in a single optical trap.

We experimentally demonstrate that optical tweezers can be used to accelerate the self-assembly of colloidal particles at a water–air interface in this Letter. The thermal flow induced by optical tweezers dominates the growth acceleration at the interface. Furthermore, optical tweezers are used to create a local growth peak at the growing front, which is used to study the preferential incorporation positions of incoming particles. The results show that the particles surfed with a strong Marangoni flow tend to fill the gap and smoothen the steep peaks. When the peak is smooth, the incoming particles incorporate the crystal homogeneously at the growing front.

We present an experimental study on opto-thermal oscillation and trapping of light absorbing particles. The oscillation is a three-dimensional motion in the solution. The particles at the lower substrate of the sample cell are driven towards the center of optical trap by the optical force. When the particles arrive at the location near the trap center, the laser heating on the particles results in a strong thermal gradient force that repels the particles to leave the focus spot. Next, the particles slow down under the viscous drag force. At last, the particles settle to the lower substrate of sample cell due to gravity, and restart the new oscillation process. For opto-thermal trapping of the absorbing particles, the particles are dispersed in a thin cell to compress the convention and enhance the viscous resistance. The particles can be trapped close to the spot due to the balance of optical and thermal gradient forces.

In this erratum, the function z(θ) in equation (7) of Opt. Lett.44, 2843 (2019)OPLEDP0146-959210.1364/OL.44.002843 has been corrected.

Optical tweezers with a low numerical aperture microscope objective is used to manipulate the microspheres at the water-air interface. In this letter, we determine the optimal optical trap for the lateral manipulation of microspheres at a water-air interface. The experimental results show that the trapping force is influenced by the expansion of the trapping beam at the back aperture of the objective. The optimal filling ratio of 0.65 is suggested for lateral optical manipulation at the water-air interface. The lateral trapping forces at the water-air interface are theoretically investigated with the ray-optics model. The numerical results show that the lateral trapping forces can be changed by shrinking the diameter of the trapping laser beam. The numerical results are in accordance with the experimental results.

One limitation on the performance of optical tweezers (abbreviated as OT) is the noise inherently present in each setup. Therefore, it is the desire to minimize and possibly eliminate the noise from the OT experiments. In this paper, a filter method based on wavelet analysis is proposed. At first we investigate the properties of OT outputs noise, and introduce the wavelet filtering method in simply. Following, we study on the OTs drift signal using different base: db4 and Haar. And also study on the signal using different filter algorithm: the soft,the hard threshold,and compulsive filter. These main conclusions based on foregoing analysis are reached: more larger the resolving scale is, more perfect the filtering effect is. The soft threshold value filtering effect is better than that of the hard threshold value filtering at the cost of calculation when the threshold value is same. The variance of the compulsive filtering is least when both the wavelet and the resolving scale are same for these filtering methods. For the compulsive filtering with same wavelets, the filtering effect of harr is better than that of db4 and the calculation of the former is fewer. Analysis the dynamic output of OT with different algorithm, it also shows that the effect of filter with the compulsive filtering is better than others. Accordingly, we found that applying the compulsive filtering with the Harr wavelet base and suitable resolving scale to the signal processing of OT outputs signal is helpful for the OT design and construction.

We report, in theory and experiment, on a novel class of controlled light capsules with nearly perfect darkness, directly employing intrinsic properties of modified Bessel-Gauss beams. These beams are able to naturally create three-dimensional bottle-shaped region during propagation as long as the parameters are properly chosen. Remarkably, the optical bottle can be controlled to demonstrate various geometries through tuning the beam parameters, thereby leading to an adjustable light capsule. We provide a detailed insight into the theoretical origin and characteristics of the light capsule derived from modified Bessel-Gauss beams. Moreover, a binary digital micromirror device (DMD) based scheme is first employed to shape the bottle beams by precise amplitude and phase manipulation. Further, we demonstrate their ability for optical trapping of core-shell magnetic microparticles, which play a particular role in biomedical research, with holographic optical tweezers. Therefore, our observations provide a new route for generating and controlling bottle beams and will widen the potentials for micromanipulation of absorbing particles, aerosols or even individual atoms., Comment: 22 pages, 8 figures

In optical tweezers, a piezo-stage (PZT) is widely used to precisely position samples for force clamp, calibrating optical trap and stretching DNA. For a trapped bead in solution, the oscillation response of PZT is vital for all kinds of applications. A coupling ratio, actual amplitude to nominal amplitude, can be calibrated by power spectral density during sinusoidal oscillations. With oscillation frequency increasing, coupling ratio decreases in both x- and y-directions, which is also confirmed by the calibration with light scattering of scanning two aligned beads on slide. Those oscillation responses are related with deformability of chamber and the intrinsic characteristics of PZT. If we take nominal amplitude as actual amplitude for sinusoidal oscillations at 50 Hz, the amplitude is overestimated ~2 times in x-direction and ~3 times in y-direction. That will lead to huge errors for subsequent calibrations.

Since RGD peptides (R: arginine; G: glycine; D: aspartic acid) are found to promote cell adhesion, they are modified at numerous materials surface for medical applications such as drug delivery and regenerative medicine. Peptide-cell surface interactions play a key role in the above applications. In this letter, we study the adhesion force between the RGD-coated bead and Hela cell surface by optical tweezes. The adhesion is dominated by the binding of α5β1 and RGD-peptide with higher adhesion probability and stronger adhesion strength compared with the adhesion of bare bead and cell surface. The binding force for a single α5β1-GRGDSP pair is determined to be 16.8 pN at a loading rate of 1.5 nN/s. The unstressed off-rate is 1.65 × 10-2 s-1 and the distance of transition state for the rigid binding model is 3.0 nm.

Optical tweezers have been used to trap and manipulate microparticles within living animals. When the optical trap is constructed with an oil-immersion objective, it suffers from spherical aberration. There have been many investigations on the influence of spherical aberration when the particles are trapped in a water medium. However, the dependence of optical force on trapping depth is still ambiguous when the trapped particles are immersed in a high refractive index medium (such as biological tissue, refractive index solution) in experiments. In this paper, the microparticles are immersed in an aqueous solution of glycerol to mimic the cells within biological tissue. As the trapping laser is focused into the specimen, spherical aberration is introduced, degrading the optical trapping performance. It is similar to trapping in water; altering the effective tube length can also compensate for the spherical aberration of the optical trap in a high refractive index medium. Finally, the cells in living mice are trapped by the optical tweezers with the help of spherical aberration compensation.

Recently, V. V. Kotlyar et al. [Opt. Lett.39, 2395 (2014)] have theoretically proposed a novel kind of three-parameter diffraction-free beam with a crescent profile, namely, the asymmetric Bessel (aB) beam. The asymmetry degree of such nonparaxial modes was shown to depend on a nonnegative real parameter c. We present a more generalized asymmetric Bessel mode in which the parameter c is a complex constant. This parameter controls not only the asymmetry degree of the mode but also the orientation of the optical crescent, and affects the energy distribution and orbital angular momentum (OAM) of the beam. As a proof of concept, the high-quality generation of asymmetric Bessel-Gauss beams was demonstrated with the super-pixel method using a digital micromirror device (DMD). We investigated the near-field properties as well as the far field features of such beams, and the experimental observations were in good agreement with the theoretical predictions. Additionally, we provided an effective way to control the beam’s asymmetry and orientation, which may find potential applications in light-sheet microscopy and optical manipulation.

The power spectrum density (PSD) has long been explored for calibrating optical tweezers stiffness. Fast Fourier Transform (FFT) based spectral estimator is typically used. This approach requires a relatively longer data acquisition time to achieve adequate spectral resolution. In this paper, an autoregressive (AR) model is proposed to obtain the Spectrum Density using a limited number of samples. According to our method, the arithmetic model has been established with burg arithmetic, and the final prediction error criterion has been used to select the most appropriate order of the AR model, the power spectrum density has been estimated based the AR model. Then, the optical tweezers stiffness has been determined with the simple calculation from the power spectrum. Since only a small number of samples are used, the data acquisition time is significantly reduced and real-time stiffness calibration becomes feasible. To test this calibration method, we study the variation of the trap stiffness as a function of the parameters of the data length and the trapping depth. Both of the simulation and experiment results have showed that the presented method returns precise results and outperforms the conventional FFT method when using a limited number of samples.

We demonstrate optical trapping of red blood cells (RBCs) in living animals by using a water immersion objective. First, the cells within biological tissue are mimicked by the particles immersed in aqueous solutions of glycerol. The optical forces depending on trapping depth are investigated when a parallel laser beam enters the water immersion objective. The results show that the optical forces vary with trapping depth, and the optimal trapping depth in aqueous solutions of glycerol (n=1.39) is 50 μm. Second, the optimal trapping depth in aqueous solutions of glycerol can be changed by altering the actual tube length of the water immersion objective. Finally, we achieved optical trapping and manipulation of RBCs in living mice.

We experimentally demonstrated Bessel-like beams utilizing digital micromirror device (DMD). DMD with images imitating the equivalent axicon can shape the collimated Gaussian beam into Bessel beam. We reconstructed the 3D spatial field of the generated beam through a stack of measured cross-sectional images. The output beams have the profile of Bessel function after intensity modulation, and the beams extend at least 50 mm while the lateral dimension of the spot remains nearly invariant. Furthermore, the self-healing property has also been investigated, and all the experimental results agree well with simulated results numerically calculated through beam propagation method. Our observations demonstrate that the DMD offers a simple and efficient method to generate Bessel beams with distinct nondiffracting and self-reconstruction behaviors. The generated Bessel beams will potentially expand the applications to the optical manipulation and high-resolution fluorescence imaging owing to the unique nondiffracting property.

The recent development of non-invasive imaging techniques has enabled the visualization of molecular events underlying cellular processes in live cells. Although microscopic objects can be readily manipulated at the cellular level, additional physiological insight is likely to be gained by manipulation of cells in vivo, which has not been achieved so far. Here we use infrared optical tweezers to trap and manipulate red blood cells within subdermal capillaries in living mice. We realize a non-contact micro-operation that results in the clearing of a blocked microvessel. Furthermore, we estimate the optical trap stiffness in the capillary. Our work expands the application of optical tweezers to the study of live cell dynamics in animals.

A symmetric Airy beam (SAB) autofocuses during free space propagation. Such autofocusing SAB is useful in optical manipulation and biomedical imaging. However, its inherently limited autofocusing property may degrade the performance of the SAB in those applications. To enhance the autofocus, a symmetric apodization mask was proposed to regulate the SAB. In combination with the even cubic phase that shapes the SAB, this even exponential function mask with an adjustable parameter regulates the contribution of different frequency spectral components to the SAB. The propagation properties of this new amplitude modulated SAB (AMSAB) were investigated both theoretically and experimentally. Simulation shows that the energy distribution and autofocusing property of an AMSAB can be adjusted by the exponential amplitude modulation. Especially, the beam energy will be more concentrated in the central lobe once the even cubic phase is modulated by the mask with a higher proportion of high-frequency spectral components. Consequently, the autofocusing property and axial gradient force of AMSABs are efficiently enhanced. The experimental generation and characterization for AMSABs were implemented by modulating the collimated beam with a phase-only spatial light modulator. The experimental results well supported the theoretical predictions. With the ability to enhance the autofocus, the proposed exponential apodization modulation will make SAB more powerful in various applications, including optical trapping, fluorescence imaging and particle acceleration.

For a triaxial ellipsoid in an optical trap with spherical aberration, the optical forces, torque and stress are analyzed using vectorial ray tracing. The torque will automatically regulate ellipsoid's long axis parallel to optic axis. For a trapped ellipsoid with principal axes in the ratio 1:2:3, the high stress distribution appears in x-z plane. And the optical force at x-axis is weaker than at y-axis due to the shape size. While the ellipsoid departs laterally from trap center, the measurable maximum transverse forces will be weakened due to axial equilibrium and affected by inclined orientation. For an appropriate ring beam, the maximum optical forces are strong in three dimensions, thus, this optical trap is appropriate to trap cells for avoiding damage from laser.

Optical tweezers has been used to manipulate micro-sized particles for many years, and has been widely used in various applications. The axial trapping stiffness is one of the most important parameters to evaluate the trapping ability of an optical tweezers. In this paper, we calibrated the axial optical stiffnesses for micro-sized polystyrene spheres. When an external force was applied to particle held by an optical trap, the particle was displaced from the trap center by an amount proportional to the applied force. We displaced the particle from the trap center by applying triangular waves of varying velocity, and the varying velocity was obtained by altering the frequency of the triangular waves. In this case the particle has two balance position distributed at two-side of the trap center. The calibration of the axial position was critical to the measurement of axial optical stiffness. In this paper, the axial displacement between the balance position and the trap center was calibrated with image information entropy signals. According to Stokes Law, when the axial displacement of the particle relative to the external force was known, the axial optical stiffness can be measured, and this method was known as viscous drag method. The stiffnesses for a 2μm-diameter at different trapped depth were measured. Typical values for axial optical stiffness of our optical tweezers were between 4.0 and 7.5 pN/μm when the laser power was 35mW. Dependence of axial optical trapping stiffness on the diameter of the particles was measured with viscous drag method. At last, the origin of the measurement error was discussed.

We introduce a corrected sinusoidal-wave drag force method (SDFM) into optical tweezers to calibrate the trapping stiffness of the optical trap and conversion factor (CF) of photodetectors. First, the theoretical analysis and experimental result demonstrate that the correction of SDFM is necessary, especially the error of no correction is up to 11.25% for a bead of 5 μm in diameter. Second, the simulation results demonstrate that the SDFM has a better performance in the calibration of optical tweezers than the triangular-wave drag force method (TDFM) and power spectrum density method (PSDM) at the same signal-to-noise ratio or trapping stiffness. Third, in experiments, the experimental standard deviations of calibration of trapping stiffness and CF with the SDFM are about less than 50% of TDFM and PSDM especially at low laser power. Finally, the experiments of stretching DNA verify that the in situ calibration with the SDFM improves the measurement stability and accuracy.

We propose a novel technique to directly transform a linearly polarized Gaussian beam into vector-vortex beams with various spatial patterns. Full high-quality control of amplitude and phase is implemented via a Digital Micro-mirror Device (DMD) binary holography for generating Laguerre-Gaussian, Bessel-Gaussian, and helical Mathieu–Gaussian modes, while a radial polarization converter (S-waveplate) is employed to effectively convert the optical vortices into cylindrically polarized vortex beams. Additionally, the generated vector-vortex beams maintain their polarization symmetry after arbitrary polarization manipulation. Due to the high frame rates of DMD, rapid switching among a series of vector modes carrying different orbital angular momenta paves the way for optical microscopy, trapping, and communication.

Optical trapping of core-shell magnetic microparticles is experimentally demonstrated by using cylindrical vector beams. Second, we investigate the optical trapping efficiencies. The results show that radially and azimuthally polarized beams exhibit higher axial trapping efficiencies than the Gaussian beam. Finally, a trapped particle is manipulated to kill a cancer cell. The results make possible utilizing magnetic particles for optical manipulation, which is an important advantage for magnetic particles as labeling agent in targeted medicine and biological analysis.

The Brownian motion of a polystyrene bead trapped in a time-sharing optical tweezers (TSOT) is numerically simulated by adopting Monte-Carlo technique. By analyzing the Brownian motion signal, the effective stiffness of a TSOT is acquired at different switching frequencies. Simulation results confirm that for a specific laser power and duty ratio, the effective stiffness varies with the frequency at low frequency range, while at high frequency range it keeps constant. Our results reveal that the switching frequency can be used to control the stability of time-sharing optical tweezers in a range.

The transverse trapping forces on a dielectric sphere located at an oil-water interface are theoretically investigated with the ray-optics model. The transverse trapping forces rely on the internal property of the particle-interface system, and increase with either the decrease of three phase contact angles at the oil-water interface or the use of oil phase with low refractive index. The numerical results also show that the transverse trapping forces can be improved by either decreasing the numerical aperture of the microscope objective or shrinking the diameter of the trapping laser beam.

Birefringent particles rotate when trapped in elliptically polarized light. When an infinity corrected oil-immersion objective is used for trapping, rotation of birefringent particles in optical tweezers based on an infinity optical microscope is affected by the spherical aberration at the glass-water interface. The maximum rotation rate of birefringent particles occurs close to the coverslip, and the rotation rate decreases dramatically as the trapped depth increases. We experimentally demonstrate that spherical aberration can be compensated by using a finite-distance-corrected objective to trap and rotate the birefringent particles. It is found that the trapped depth corresponding to the maximum rotation rate is 50 microm, and the rotation rates at deep trapped depths are improved.