Optical trapping of gold microparticles using linearly polarized, centrally obstructed Gaussian beams.

Metallic microparticles larger than the illumination wavelength are commonly considered poor optical trapping candidates due to their high extinction coefficient. This paper presents a numerical and experimental study on the three-dimensional (3D) trapping of gold microparticles using a centrally obstructed Gaussian beam based on the T-matrix method. The range of particle size for stable optical trapping is determined. For the trapping numerical aperture of 1.32 and illumination wavelength of 1.064 µm, numerical analysis proves that 3D trapping of gold microparticles with a radius bigger than 1.0 µm can be readily achieved. By imprinting a digital lens to the spatial light modulator, we slightly defocus the centrally obstructed Gaussian beam to shift the trapping location to the focal plane for clear observation. Experimental results demonstrate stable trapping of gold microparticles with a radius greater than 1.4 µm at high-power illumination, agreeing well with the theoretical predictions. The presented work should be of interest to the community applying metallic microparticles to relevant research.

Zero-order free holographic optical tweezers.

Holographic optical tweezers (HOTs) use spatial light modulators (SLM) to modulate light beams, thereby enabling the dynamic control of optical trap arrays with complex intensity and phase distributions. This has provided exciting new opportunities for cell sorting, microstructure machining, and studying single molecules. However, the pixelated structure of the SLM will inevitably bring up the unmodulated zero-order diffraction possessing an unacceptably large fraction of the incident light beam power. This is harmful to optical trapping because of the bright, highly localized nature of the errant beam. In this paper and to address this issue, we construct a cost-effective, zero-order free HOTs apparatus, thanks to a homemade asymmetric triangle reflector and a digital lens. As there is no zero-order diffraction, the instrument performs excellently in generating complex light fields and manipulating particles.

Rapid, artifact-reduced, image reconstruction for super-resolution structured illumination microscopy.

Super-resolution structured illumination microscopy (SR-SIM) is finding increasing application in biomedical research due to its superior ability to visualize subcellular dynamics in living cells. However, during image reconstruction artifacts can be introduced and when coupled with time-consuming postprocessing procedures, limits this technique from becoming a routine imaging tool for biologists. To address these issues, an accelerated, artifact-reduced reconstruction algorithm termed joint space frequency reconstruction-based artifact reduction algorithm (JSFR-AR-SIM) was developed by integrating a high-speed reconstruction framework with a high-fidelity optimization approach designed to suppress the sidelobe artifact. Consequently, JSFR-AR-SIM produces high-quality, super-resolution images with minimal artifacts, and the reconstruction speed is increased. We anticipate this algorithm to facilitate SR-SIM becoming a routine tool in biomedical laboratories.

Multi-color structured illumination microscopy for live cell imaging based on the enhanced image recombination transform algorithm.

Structured illumination microscopy (SIM) has attracted considerable interest in super-resolution, live-cell imaging because of its low light dose and high imaging speed. Obtaining a high-quality reconstruction image in SIM depends on the precise determination of the parameters of the fringe illumination pattern. The image recombination transform (IRT) algorithm is superior to other algorithms in obtaining the precise initial phase without any approximation, which is promising to provide a considerable solution to address the difficulty of initial phase estimation at low-modulation-depth conditions. However, the IRT algorithm only considers a phase shift of π∕2, which limits its applications in general scenarios. In this letter, we present a general form of IRT algorithm suitable for arbitrary phase shifts, providing a powerful tool for parameter estimation in low signal-to-noise cases. To demonstrate the effectiveness of the enhanced IRT algorithm, we constructed a multicolor, structured illumination microscope and studied at super-resolution, the cargo traffic in HRPE cells, and monitored the movement of mitochondrial structures and microtubules in COS-7 cells. The custom SIM system using the enhanced IRT algorithm allows multicolor capability and a low excitation intensity fluorescence imaging less than 1 W/cm2. High-quality super-resolution images are obtained, which demonstrates the utility of this approach in imaging in the life sciences.

Quantitative proteomics implicates YggT in streptomycin resistance in Salmonella enterica serovar Enteritidis.

OBJECTIVES: To identify proteins that may be associated with antibiotic resistance in the multidrug-resistant Salmonella enterica D14, by constructing proteomic profiles using mass spectrometry-based label-free quantitative proteomics (LFQP). RESULTS: D14 was cultured with four antibiotics (ampicillin, nalidixic acid, streptomycin, and tetracycline) separately. Subsequently, the findings from an equal combination of the four cultures were compared with the profile of sensitive S. enterica 104. 2255 proteins, including 149 differentially up-regulated proteins, were identified. Many of these up-regulated proteins were associated with flagellar assembly and chemotaxis, two-component system, amino acid metabolism, ß-lactam resistance, and transmembrane transport. A subset of 10 genes was evaluated via quantitative real-time PCR (qPCR), followed by the construction of cheR, fliS, fliA, arnA, and yggT deletion mutants. Only the yggT-deleted D14 mutant showed decrease in streptomycin resistance, whereas the other deletions had no effect. Furthermore, complementation of yggT and the overexpression of yggT in S. enterica ATCC 14028 increased the streptomycin resistance. Additionally, spot dilution assay results confirmed that Salmonella strains, harboring yggT, exhibited an advantage in the presence of streptomycin. CONCLUSIONS: The above proteomic and mutagenic analyses revealed that yggT is involved in streptomycin resistance in S. enterica.

Hybrid multifocal structured illumination microscopy with enhanced lateral resolution and axial localization capability.

Super-resolution (SR) fluorescence microscopy that breaks through the diffraction barrier has drawn great interest in biomedical research. However, obtaining a high precision three-dimensional distribution of the specimen in a short time still remains a challenging task for existing techniques. In this paper, we propose a super-resolution fluorescence microscopy with axial localization capability by combining multifocal structured illumination microscopy with a hybrid detection PSF composed of a Gaussian PSF and a double-helix PSF. A modified reconstruction scheme is presented to accommodate the new hybrid PSF. This method can not only recover the lateral super-resolution image of the specimen but also retain the specimen's depth map within a range of 600 nm with an axial localization precision of 20.8 nm. The performance of this approach is verified by testing fluorescent beads and tubulin in 293-cells. The developed microscope is well suited for observing the precise 3D distribution of thin specimens.

Rapid tilted-plane Gerchberg-Saxton algorithm for holographic optical tweezers.

Benefitting from the development of commercial spatial light modulator (SLM), holographic optical tweezers (HOT) have emerged as a powerful tool for life science, material science and particle physics. The calculation of computer-generated holograms (CGH) for generating multi-focus arrays plays a key role in HOT for trapping of a bunch of particles in parallel. To realize dynamic 3D manipulation, we propose a new tilted-plane GS algorithm for fast generation of multiple foci. The multi-focal spots with a uniformity of 99% can be generated in a tilted plane. The computation time for a CGH with 512×512 pixels is less than 0.1 second. We demonstrated the power of the algorithm by simultaneously trapping and rotating silica beads with a 7×7 spots array in three dimensions. The presented algorithm is expected as a powerful kernel of HOT.

Simultaneous optical trapping and imaging in the axial plane: a review of current progress.

Optical trapping has become a powerful tool in numerous fields such as biology, physics, chemistry, etc. In conventional optical trapping systems, trapping and imaging share the same objective lens, confining the region of observation to the focal plane. For the capture of optical trapping processes occurring in other planes, especially the axial plane (the one containing the z-axis), many methods have been proposed to achieve this goal. Here, we review the methods of acquiring the axial-plane information from which axial plane trapping is observed and discuss their advantages and limitations. To overcome the limitations existing in these methods, we developed an optical tweezers system that allows for simultaneous optical trapping and imaging in the axial plane. The versatility and usefulness of the system in axial-plane trapping and imaging are demonstrated by investigating its trapping performance with various optical fields, including Bessel, Airy, and snake-like beams. The potential applications of the reported technique are suggested to several research fields, including optical pulling, longitudinal optical binding, tomographic phase microscopy (TPM), and super-resolution microscopy.

Direct observation and characterization of optical guiding of microparticles by tightly focused non-diffracting beams.

Due to the propagation-invariant and self-healing properties, nondiffracting beams are highly attractive in optical trapping. However, little attention has been paid to investigating optical guiding of microparticles in nondiffracting beams generated by high-numerical-aperture (NA) optics with direct visualization. In this letter, we report a technique for direct observation and characterization of optical guiding of microparticles in a tight focusing system. With this technique, we observed a parabolic particle guiding trajectory with a longitudinal distance of more than 100µm and a maximal lateral deviation of 20 µm when using Airy beams. We also realized the tilted-path transport of microparticles with controllable guiding direction using tilted zeroth-order quasi-Bessel beams. For an NA of the focusing lens equal to 0.95, we achieved the optical guiding of microparticles along a straight path with a tilt angle of up to 18.8° with respect to the optical axis over a distance of 300 µm. Importantly, quantitative measurement of particle's motion was readily accessed by measuring the particle's position and velocity during the transport process. The reported technique for direct visualization and characterization of the guided particles will find its potential applications in optical trapping and guiding with novel nondiffracting beams or accelerating beams.

Chemerin-induced angiogenesis and adipogenesis in 3 T3-L1 preadipocytes is mediated by lncRNA Meg3 through regulating Dickkopf-3 by sponging miR-217.

PURPOSE: Obesity is often caused by the excess adipogenesis and regulated by long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). We performed this study to investigate the influence of Meg3 expression on adipogenesis and also the Meg3/miR-217/Dkk3 axis-mediated molecular mechanism in adipogenesis and angiogenesis. METHODS: 3 T3-L1 preadipocytes were incubated with chemerin and transfected with Meg3-overexpressing (OE-Meg3) and Dkk3-overexpressing (OE-Dkk3) plasmids, siRNAs, and miR-217 mimics, inhibitor and scrambled sequences for 48 h or 72 h. The changes in cell proliferation, adipogenesis and angiogenesis ability in 3 T3-L1 preadipocytes was detected by using the corresponding assay. The expressions of related proteins were detected via western blot. RESULTS: Chemerin decreased miR-217 expression and increased Meg3 expression, meanwhile promoted the proliferation, adipogenesis and angiogenesis in 3 T3-L1 preadipocytes. Besides, OE-Meg3 exerted the synergistic effect on 3 T3-L1 preadipocytes when co-treated with chemerin. The target interactions between Meg3 and miR-217 as well as between miR-217 and Dkk3 were validated using dual-luciferase reporter system. SiMeg3 antagonized chemerin-induced changes, while the addition of miR-217 inhibitor attenuated siMeg3-induced changes in 3 T3-L1 preadipocytes. The proliferation, adipogenesis and angiogenesis in 3 T3-L1 preadipocytes were suppressed by miR-217 mimics, while promoted by the OE-Dkk3 Chemerin promoted the expression of fatty acid binding protein 4 and vascular endothelial growth factor (VEGF) proteins, and decreased the expression of cyclin D1, c-Myc, and ß-catenin proteins. Meanwhile, these effects were further enhanced by OE-Meg3 or OE-Dkk3. However, the transfection of siMeg3, or miR-217 mimics, or siDkk3 reversed the previous changes. CONCLUSIONS: Meg3/miR-217/Dkk3 induced adipogenesis and angiogenesis in 3 T3-L1 preadipocytes via activating VEGF signaling pathway and inhibiting Wnt/ß-catenin signaling pathway.

Multi-view object topography measurement with optical sectioning structured illumination microscopy.

Various optical instruments have been developed for three-dimensional (3D) surface topography, including the white light interference, reflectance confocal microscopes, and digital holographic microscopes, etc. However, the steep local slope of objects may cause the light to be reflected in a way that it will not be captured by the objective lens because of the finite collection angle of the objective. To solve this "shadow problem," we report a method to enlarge the collection angle range of optical sectioning structured illumination microscopy by capturing sectioned images of the objects from multiple angle of views. We develop a multi-view image fusion algorithm to reconstruct a single 3D image. Using this method, we detect previously invisible details whose slopes are beyond the collection angle of the objective. The proposed approach is useful for height map measurement and quantitative analyses in a variety of fields, such as biology, materials science, microelectronics, etc.

FSP1 promotes the biofunctions of adventitial fibroblast through the crosstalk among RAGE, JAK2/STAT3 and Wnt3a/ß-catenin signalling pathways.

Emerging evidence indicates that fibroblast-specific protein 1 (FSP1) provides vital effects in cell biofunctions. However, whether FSP1 influences the adventitial fibroblast (AF) and vascular remodelling remains unclear. Therefore, we investigated the potential role and action mechanism of FSP1-mediated AF bioactivity. AFs were cultured and stimulated with FSP1 and siRNA-FSP1 in vitro. Viability assays demonstrated that siRNA-FSP1 counteracted AFs proliferative, migratory and adherent abilities enhanced with FSP1. Flow cytometry revealed that FSP1 increased AFs number in S phase and decreased cellular apoptosis. Contrarily, siRNA-FSP1 displayed the contrary results. RT-PCR, Western blotting and immunocytochemistry showed that FSP1 synchronously up-regulated the expression of molecules in RAGE, JAK2/STAT3 and Wnt3a/ß-catenin pathways and induced a proinflammatory cytokine profile characterized by high levels of MCP-1, ICAM-1 and VCAM-1. Conversely, FSP1 knockdown reduced the expression of these molecules and cytokines. The increased number of autophagosomes in FSP1-stimulated group and fewer autophagic corpuscles in siRNA-FSP1 group was observed by transmission electron microscope (TEM). Autophagy-related proteins (LC3B, beclin-1 and Apg7) were higher in FSP1 group than those in other groups. Conversely, the expression of p62 protein was shown an opposite trend of variation. Therefore, these pathways can promote AFs bioactivity, facilitate autophagy and induce the expression of the proinflammatory cytokines. Contrarily, siRNA-FSP1 intercepts the crosstalk of these pathways, suppresses AF functions, restrains autophagy and attenuates the expression of the inflammatory factors. Our findings indicate that crosstalk among RAGE, STAT3/JAK2 and Wnt3a/ß-catenin signalling pathways may account for the mechanism of AF functions with the stimulation of FSP1.

Generation of a double-ring perfect optical vortex by the Fourier transform of azimuthally polarized Bessel beams.

The perfect optical vortex (POV), the ring size being independent of its topological charge, has found potential applications in optical tweezers and optical communications. In this Letter, we report a new kind of POV, termed as double-ring POV (DR-POV), whose diameters of the two rings are independent of topological charge. We theoretically demonstrate that such a vortex is the Fourier transform of an azimuthally polarized Bessel beam. Experimental results agree well with theoretical prediction. We further investigate the vortex nature of the DR-POV through an interferometric method, showing that the two rings of the vortex have the same topological charge value (magnitude and sign). The specular properties of the DR-POV may find application in optical tweezers, such as trapping and rotating of low-refractive-index particles in the dark region between the two rings.

Real-time optical manipulation of particles through turbid media.

Complex diffusive scattering media pose significant challenges for light focusing as well as optical imaging to be implemented in practice. Recently, it has been demonstrated that the wavefront shaping technique can be applied to realize focusing and imaging through scattering medium. Here we report dynamic optical manipulation of particles through turbid media by employing the interleaved segment wavefront correction method, which is an improved genetic algorithm providing faster convergence speed and higher peak to background ratio. Manipulating micro-beads behind a scattering medium along both one and two dimensional predesigned trajectories in real time has been successfully demonstrated.

Aberration correction method based on double-helix point spread function.

Point spread function (PSF) engineering has met with lots of interest in various optical imaging techniques, including super-resolution microscopy, microparticle tracking, and extended depth-of-field microscopy. The intensity distributions of the modified PSFs often suffer from deteriorations caused by system aberrations, which greatly degrade the image contrast, resolution, or localization precision. We present an aberration correction method using a spiral-phase-based double-helix PSF as an aberration indicator, which is sensitive and quantitatively correlated to the spherical aberration, coma, and astigmatism. Superior to the routine iteration-based correction methods, the presented approach is iteration-free and the aberration coefficients can be directly calculated with the measured parameters, relieving the computing burden. The validity of the method is verified by both examining the intensity distribution of the conventional Gaussian PSF in three dimensions and observing muntjac skin fibroblast cells. This iteration-free correction method has a potential application in PSF engineering systems equipped with a spatial light modulator.

Aberration correction in holographic optical tweezers using a high-order optical vortex: publisher's note.

This publisher's note identifies an error in the author affiliations of Appl. Opt.57, 3618 (2018)APOPAI0003-693510.1364/AO.57.003618.

Aberration correction in holographic optical tweezers using a high-order optical vortex.

Holographic optical tweezers are a powerful optical trapping and manipulation tool in numerous applications such as life science and colloidal physics. However, imperfections in the spatial light modulator and optical components of the system will introduce detrimental aberrations to the system, thereby degrading the trapping performance significantly. To address this issue, we develop an aberration correction technique by using a high-order vortex as the correction metric. The optimal Zernike polynomial coefficients for quantifying the system aberrations are determined by comparing the distorted vortex and the ideal one. Efficiency of the proposed method is demonstrated by comparing the optical trap intensity distribution, trap stiffness, and particle dynamics in a Gaussian trap and an optical vortex trap, before and after aberration corrections.

Rotating of low-refractive-index microparticles with a quasi-perfect optical vortex.

Low-refractive-index microparticles, such as hollow microspheres, have shown great significance in some applications, such as biomedical sensing and targeted drug delivery. However, optical trapping and manipulation of low-refractive-index microparticles are challenging, owing to the repelling force exerted by typical optical traps. In this paper, we demonstrated optical trapping and rotating of large-sized low-refractive-index microparticles by using quasi-perfect optical vortex (quasi-POV) beams, which were generated by Fourier transform of high-order quasi-Bessel beams. Numerical simulation was carried out to characterize the focusing property of the quasi-POV beams. The dynamics of low-refractive-index microparticles in the quasi-POV with various topological charges was investigated in detail. To improve the trapping and rotating performances of the vortex, a point trap was introduced at the center of the ring. Experimental results showed that the quasi-POV was preferable for manipulation of large-sized low-refractive-index microparticles, with its control of the particles' rotating velocity dependent only on the topological charge due to the unchanged orbital radius.

Three-dimensional characterization of tightly focused fields for various polarization incident beams.

Tightly focused vectorial optical beams have found extensive applications in variety of technical fields like single-molecule detection, optical tweezers, and super-resolution optical microscopy. Such applications require an accurate measurement and manipulation of focal optical fields. We have developed a compact instrument (with dimensions of 35 × 35 × 30 cm3) to rapidly measure the intensity distribution in three dimensions of the focused fields of vectorial beams and any other incident beams. This instrument employs a fluorescent nanoparticle as a probe to scan the focal region to obtain a high spatial resolution of intensity distribution. It integrates a liquid-crystal spatial light modulator to allow for tailoring the point spread function of the optical system, making it a useful tool for multi-purpose and flexible research. The robust applicability of the instrument is verified by measuring the 3D intensity distributions of focal fields of various polarization and wavefront modulated incident beams focused by a high NA (=1.25) objective lens. The minimal data acquisition time achievable in the experiment is about 8 s for a scanning region of 3.2 × 3.2 µm2 (512 × 512 pixels). The measured results are in good agreement with those predicted by the vectorial diffraction theory.

Interleaved segment correction achieves higher improvement factors in using genetic algorithm to optimize light focusing through scattering media.

Focusing and imaging through scattering media has been proved possible with high resolution wavefront shaping. A completely scrambled scattering field can be corrected by applying a correction phase mask on a phase only spatial light modulator (SLM) and thereby the focusing quality can be improved. The correction phase is often found by global searching algorithms, among which Genetic Algorithm (GA) stands out for its parallel optimization process and high performance in noisy environment. However, the convergence of GA slows down gradually with the progression of optimization, causing the improvement factor of optimization to reach a plateau eventually. In this report, we propose an interleaved segment correction (ISC) method that can significantly boost the improvement factor with the same number of iterations comparing with the conventional all segment correction (ASC) method. In the ISC method, all the phase segments are divided into a number of interleaved groups; GA optimization procedures are performed individually and sequentially among each group of segments. The final correction phase mask is formed by applying correction phases of all interleaved groups together on the SLM. The ISC method has been proved significantly useful in practice because of its ability to achieve better improvement factors when noise is present in the system. We have also demonstrated that the imaging quality is improved as better correction phases are found and applied on the SLM. Additionally, the ISC method lowers the demand of dynamic ranges of detection devices. The proposed method holds potential in applications, such as high-resolution imaging in deep tissue.