Simultaneous Manipulation Research Articles

Broadband Polarization-Conversion Metasurface for a Cassegrain Antenna with High Polarization Purity

Though polarization-conversion (PC) metasurfaces have been thoroughly explored in the microwave and optical regimes, they have seldom been used in everyday applications, such as improving the overall performance of antennas for wireless communication. This study not only focuses on the strategy for designing an efficient PC metasurface, but also applies it to construct a high-performance Cassegrain antenna. This metasurface is capable of simultaneous polarization manipulation and arbitrary phase tailoring, while the antenna has the advantages of low profile, high directivity, broad bandwidth, and high polarization purity.

Abstract 1759: Studying T-cell force activation and DNA repair dynamics at the single-molecule level with optical tweezers and correlated fluorescence microscopy

Abstract DNA repair, the collection of highly regulated mechanisms by which a cell identifies and repairs DNA damage, remains one of the most essential processes of human life. Without DNA repair mechanisms cells lose the ability to transcribe important regions of their genome, resulting in harmful mutations, which could eventually jeopardize cellular wellbeing. Sources of DNA damage include double-stranded breaks and DNA intra- and interstrand crosslinks, which can ultimately become malignant tumors, leading to cancer. To study DNA repair, single-molecule studies have proven to greatly enhance understanding at the molecular level. However, it is often challenging to obtain adequate sensitivity and resolution, as well as biological conditions mimicking in vivo environments. The C-Trap™ system allows for realtime visualization of the interaction between DNA and DNA repair proteins, under biologically relevant conditions with high spatial and temporal resolution. The C-Trap™ is the only instrument that integrates optical tweezers, confocal/STED microscopy, and an advanced microfluidics system in a truly correlated manner. It enables live, simultaneous and correlative visualization and manipulation of molecular interactions with sub-picoNewton (pN) force resolution and a kilo-Hz (to mega-Hz) temporal resolution. Previously we have presented applications in: protein (un)folding and conformational changes; DNA-protein interactions and genome modifications; effects of mechanical stress on DNA/RNA structure; motility of cytoskeletal molecular motors; and protein droplet and aggregation dynamics. By culturing or adhering cells to the surface of the flow chamber, single-molecule cell-based studies can also be performed with the C-Trap. Cell-surface bound mechano-receptors can be activated by forming tethers with an optically trapped particle and then creating a controlled tension on the molecule. Correlated fluorescent imaging can elucidate internal protein cascades and other similar reactions to the applied force. Here, we present our experiments visualizing and quantifying the mechanisms of DNA repair and the force activation of T-cells using the C-Trap system. These experiments show that the technological advances in hybrid single-molecule methods can be turned into an easy-to-use and stable instrument that opens up new venues in many research areas. Citation Format: Ali Raja, Leif Anderson, Willem Peutz, Andrea Candelli, Gerrit Sitters. Studying T-cell force activation and DNA repair dynamics at the single-molecule level with optical tweezers and correlated fluorescence microscopy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1759.

High-throughput multicolor optogenetics in microwell plates.

Optogenetic probes can be powerful tools for dissecting complexity in cell biology, but there is a lack of instrumentation to exploit their potential for automated, high-information-content experiments. This protocol describes the construction and use of the optoPlate-96, a platform for high-throughput three-color optogenetics experiments that allows simultaneous manipulation of common red- and blue-light-sensitive optogenetic probes. The optoPlate-96 enables illumination of individual wells in 96-well microwell plates or in groups of wells in 384-well plates. Its design ensures that there will be no cross-illumination between microwells in 96-well plates, and an active cooling system minimizes sample heating during light-intensive experiments. This protocol details the steps to assemble, test, and use the optoPlate-96. The device can be fully assembled without specialized equipment beyond a 3D printer and a laser cutter, starting from open-source design files and commercially available components. We then describe how to perform a typical optogenetics experiment using the optoPlate-96 to stimulate adherent mammalian cells. Although optoPlate-96 experiments are compatible with any plate-based readout, we describe analysis using quantitative single-cell immunofluorescence. This workflow thus allows complex optogenetics experiments (independent control of stimulation colors, intensity, dynamics, and time points) with high-dimensional outputs at single-cell resolution. Starting from 3D-printed and laser-cut components, assembly and testing of the optoPlate-96 can be accomplished in 3-4 h, at a cost of ~$600. A full optoPlate-96 experiment with immunofluorescence analysis can be performed within ~24 h, but this estimate is variable depending on the cell type and experimental parameters.

A flexible geometry for panoramic visual and optogenetic stimulation during behavior and physiology

BackgroundTo study visual processing, it is necessary to precisely control visual stimuli while recording neural and behavioral responses. It can be important to present stimuli over a broad area of the visual field, which can be technically difficult. New MethodWe present a simple geometry that can be used to display panoramic stimuli. A single digital light projector generates images that are reflected by mirrors onto flat screens that surround an animal. It can be used for behavioral and neurophysiological measurements, so virtually identical stimuli can be presented. Moreover, this geometry permits light from the projector to be used to activate optogenetic tools. ResultsUsing this geometry, we presented panoramic visual stimulation to Drosophila in three paradigms. We presented drifting contrast gratings while recording walking and turning speed. We used the same projector to activate optogenetic channels during visual stimulation. Finally, we used two-photon microscopy to record responses in direction-selective cells to drifting gratings. Comparison with Existing Method(s)Existing methods have typically required custom hardware or curved screens, while this method requires only flat back projection screens and a digital light projector. The projector generates images in real time and does not require pre-generated images. Finally, while many setups are large, this geometry occupies a 30 × 20 cm footprint with a 25 cm height. ConclusionsThis flexible geometry enables measurements of behavioral and neural responses to panoramic stimuli. This allows moderate throughput behavioral experiments with simultaneous optogenetic manipulation, with easy comparisons between behavior and neural activity using virtually identical stimuli.

Spin-Orbit Interaction of Light in Plasmonic Lattices.

In the past decade, the spin-orbit interaction (SOI) of light has been a driving force in the design of metamaterials, metasurfaces, and schemes for light-matter interaction. A hallmark of the spin-orbit interaction of light is the spin-based plasmonic effect, converting spin angular momentum of propagating light to near-field orbital angular momentum. Although this effect has been thoroughly investigated in circular symmetry, it has yet to be characterized in a noncircular geometry, where whirling, periodic plasmonic fields are expected. Using phase-resolved near-field microscopy, we experimentally demonstrate the SOI of circularly polarized light in nanostructures possessing dihedral symmetry. We show how interaction with hexagonal slits results in four topologically different plasmonic lattices, controlled by engineered boundary conditions, and reveal a cyclic nature of the spin-based plasmonic effect which does not exist for circular symmetry. Finally, we calculate the optical forces generated by the plasmonic lattices, predicting that light with mere spin angular momentum can exert torque on a multitude of particles in an ordered fashion to form an optical nanomotor array. Our findings may be of use in both biology and chemistry, as a means for simultaneous trapping, manipulation, and excitation of multiple objects, controlled by the polarization of light.

CRISPR/Cas9-RNA interference system for combinatorial metabolic engineering of Saccharomyces cerevisiae.

The yeast Saccharomyces cerevisiae is widely used in industrial biotechnology for the production of fuels, chemicals, food ingredients, food and beverages, and pharmaceuticals. To obtain high‐performing strains for such bioprocesses, it is often necessary to test tens or even hundreds of metabolic engineering targets, preferably in combinations, to account for synergistic and antagonistic effects. Here, we present a method that allows simultaneous perturbation of multiple selected genetic targets by combining the advantage of CRISPR/Cas9, in vivo recombination, USER assembly and RNA interference. CRISPR/Cas9 introduces a double‐strand break in a specific genomic region, where multiexpression constructs combined with the knockdown constructs are simultaneously integrated by homologous recombination.We show the applicability of the method by improving cis,cis‐muconic acid production in S. cerevisiae through simultaneous manipulation of several metabolic engineering targets.The method can accelerate metabolic engineering efforts for the construction of future cell factories.

A microfluidic platform for multi-size 3D tumor culture, monitoring and drug resistance testing

The establishment of a miniaturized platform capable of microscale control for sequential, coordinated, and throughput biomimetic tumor manipulation and analysis offer a high degree of biological and clinical relevance to cancer therapy and pharmaceutical research. In this study, we developed a microfluidic system with well-established microwell arrays and simple double-layer composition that allows massive fabrication of biomimetic multi-size 3D tumors and their simultaneous manipulation and orthogonal analysis. Serial manipulations including cell localization, array-like self-assembly/cultivation, and dynamic analysis of different types of 3D tumors, were accomplished in the microfluidic device. We demonstrated that the microfluidic platform is stable and throughput cell trapping and tumor generation with quantity uniform. Furthermore, on-chip monitoring and throughput analysis of tumor phenotypes and responses to culture and different chemotherapies were also achieved in the device. The microfluidic advancement offers a new methodological approach for the development of high-performance and multifunctional 3D tumor systems and for tissue-mimicking cancer research and therapy evaluation. This microfluidic platform, which has the capability of multiple control in time and space, holds great potential for applications in the fields of tumor biology, tissue engineering, and clinical medicine.

Engineering Ultrathin MoS2 Nanosheets Anchored on N‐Doped Carbon Microspheres with Pseudocapacitive Properties for High‐Performance Lithium‐Ion Capacitors

AbstractLithium‐ion capacitors (LICs) are emerging as one of the most advanced energy storage devices by combining the virtues of both supercapacitor and lithium‐ion battery. The key point of constructing high‐performance LICs is to balance the electrochemical kinetics and capacity mismatch between battery‐type anode and capacitive‐type cathode materials. Herein, a strategy is presented for simultaneous manipulation of a MoS2/N‐doped carbon microspheres anode and hierarchical porous carbon cathode by using polyimide precursor. Owing to the fast lithium diffusion rate, high pseudocapacitive behavior, and expanding the interlayer of the MoS2 composite network architecture, the material can achieve excellent rate capacity and cyclic stability. Hierarchical porous carbon has an ultrahigh specific surface area and superior capacitive behavior. A high‐performance LIC is successfully constructed by using the superior anode and cathode materials. The device can deliver a maximum energy density of 120 Wh kg−1 and keep the capacity retention of 85.5% after 4000 cycles, revealing the competition in advanced energy storage devices. Accordingly, the simultaneous manipulation of metal sulfide and hierarchical porous carbon by the same precursor can be used toward fabricating other ideal electrode structures for energy storage.

Vision-Based Surgical Suture Looping Through Trajectory Planning for Wound Suturing

Robot-assisted surgery has revolutionized the field of surgery over the past few decades. Despite many successes, achieving full automation in these surgeries remains a challenging task. In this paper, a dynamic approach is proposed to automate knot tying with an in-house robot vision system. Through efficient path planning and coordination between two grippers, the workspace required for constructing the suture loop can be reduced while issues such as suture slippage and collisions between instruments can be eliminated. Visual images were employed to monitor the two grippers in real time and their positions were evaluated using transformation matrices obtained experimentally. A linear quadratic control scheme was applied to optimize the tracking performance of the two grippers. From the experiments, this visual evaluation method can achieve a position accuracy of 1 mm in the workspace. The proposed algorithm was evaluated and automatic suture looping operation was successfully performed in all six trials. Different parameters in the control scheme were also examined by introducing external impulse disturbances during the knot-tying process. This proposed knot-tying approach demonstrates a simple and efficient way to construct a suture knot in a minimal workspace. Note to Practitioners —A surgical operation usually takes several hours to complete, which is a test of surgeon’s endurance. To better assist surgeons, several repetitive surgical tasks can be performed automatically with a robotic system. This paper presents a new method for robot-assisted surgical knot tying in a confined environment. Through simultaneous manipulation and trajectory planning of the two surgical grippers, suture loops were constructed while keeping the suture in tension. Images were employed to guide the gripers in completing the knot-tying process with high precision. Experiments were conducted to demonstrate the automated suture looping operation with the system and the proposed method. The robustness of the method was also examined by introducing impulse disturbances and occlusions during the experiments. This proposed method does not require complicated hardware setup, allowing easy implementations on various surgical systems.

Synthesis of Colloidal SU-8 Polymer Rods Using Sonication.

The bulk synthesis of fluorescent colloidal SU-8 polymer rods with tunable dimensions is described. The colloidal SU-8 rods are prepared by shearing an emulsion of SU-8 polymer droplets and then exposing the resulting non-Brownian rods to ultrasonic waves, which breaks them into colloidal rods with typical lengths of 3.5-10 µm and diameters of 0.4-1 µm. The rods are stable in both aqueous and apolar solvents, and by varying the composition of apolar solvent mixtures both the difference in refractive index and mass density between particles and solvent can be independently controlled. Consequently, these colloidal SU-8 rods can be used in both 3D confocal microscopy and optical trapping experiments while carefully tuning the effect of gravity. This is demonstrated by using confocal microscopy to image the liquid crystalline phases and the isotropic-nematic interface formed by the colloidal SU-8 rods and by optically trapping single rods in water. Finally, the simultaneous confocal imaging and optical manipulation of multiple SU-8 rods in the isotropic phase is shown.

Feasibility of using a fully immersive virtual reality system for kinematic data collection

Commercially-available Virtual Reality (VR) systems have the potential to be effective tools for simultaneous visual manipulation and kinematic data collection. Previously, these systems have been integrated with research-grade motion capture systems to provide both functionalities; however, they are yet to be used as stand-alone systems for kinematic data collection. The present study aimed to validate the HTC VIVE VR system for kinematic data collection by evaluating the accuracy of its position and orientation signals. The VIVE controller and tracker were each compared to a Polhemus Liberty magnetic tracking system sensor for angular and translational measurement error and signal drift. A sensor from each system was mounted to opposite ends of a rigid segment which was driven through fifty rotations and fifty translations. Mean angular errors for both the VIVE tracker and controller were below 0.4°. Mean translational error for both sensors was below 3 mm. Drift in the Liberty signal components was consistently lower than drift in VIVE components. However, all mean rotational drift measures were below 0.1° and all mean translational measures were below 0.35 mm. These data indicate that the HTC VIVE system has the potential to be a valid and reliable means of kinematic data collection. However, further investigation is necessary to determine the VIVE’s suitability for capturing extremely minute or high-volume movements.

Millimeter-structured nondiffracting surface beams

A new type of scalar nondiffracting beam is presented whose main feature is the freedom on the choice of a two-dimensional spatial intensity pattern dependent upon the longitudinal coordinate. Examples of such millimeter-structured surface beams are provided over Cartesian and cylindrical planes, and, motivated by the idea of designing beams capable of simultaneous manipulation, guidance, and/or trapping of micro- and millimeter-size particles, here we also derive analytical expressions for their beam shape coefficients in the framework of the generalized Lorenz–Mie theory, a first step toward the analysis of light–matter interactions with spherical particles from such light fields.

Simultaneous manipulation of microparticles in multiple planes with the Thue–Morse zone plate beam

We demonstrate optical manipulation with an optical beam generated by a Thue–Morse zone plate (TMZP) in this paper. The results show that the generated TMZP beam can simultaneously manipulate microparticles positioned in multiple focal planes of the beam, owing to the self-reconstruction property and the symmetry foci with the same intensity along the optical axis. The TMZP beam will be promising in the three-dimensional optical tweezers.

Intensiometric biosensors visualize the activity of multiple small GTPases in vivo

Ras and Rho small GTPases are critical for numerous cellular processes including cell division, migration, and intercellular communication. Despite extensive efforts to visualize the spatiotemporal activity of these proteins, achieving the sensitivity and dynamic range necessary for in vivo application has been challenging. Here, we present highly sensitive intensiometric small GTPase biosensors visualizing the activity of multiple small GTPases in single cells in vivo. Red-shifted sensors combined with blue light-controllable optogenetic modules achieved simultaneous monitoring and manipulation of protein activities in a highly spatiotemporal manner. Our biosensors revealed spatial dynamics of Cdc42 and Ras activities upon structural plasticity of single dendritic spines, as well as a broad range of subcellular Ras activities in the brains of freely behaving mice. Thus, these intensiometric small GTPase sensors enable the spatiotemporal dissection of complex protein signaling networks in live animals.

Multisegmented Metallic Nanorods: Sub-10 nm Growth, Nanoscale Manipulation, and Subwavelength Imaging.

Multisegmented metallic nanorods (MS-M-NRs) have attracted increasing attention thanks to their integrated structures and complex functions. The integration of nanoscale segments in 1D enables maximum exposure of each segment and enhanced interaction between adjacent segments. Such structural integration will induce functional complexity in the nanorods, leading to superior properties for the individual components. Herein, recent progress on the development of MS-M-NRs is reviewed. Their precise fabrication, nanoscale manipulation, and subwavelength imaging, as well as simultaneous manipulation and imaging are discussed, respectively. Specifically, precise fabrication of MS-M-NRs focuses on porous anodic alumina (PAA) templated electrodeposition, which enables sub-10 nm growth of the segments and their interfaces/fronts. Nanoscale manipulation of MS-M-NRs introduces the fundamental methods that are employed for delicate movement control on the nanorods through internal or external stimulations. Subwavelength imaging of MS-M-NRs highlights the achievements on identification and location of constituent nanoscale segments/gaps based on their differences and interactions. Simultaneous manipulation and imaging of MS-M-NRs addresses the significance and potential applications of the nanorods with magnetic-plasmonic dual modulation. The development of MS-M-NRs will greatly contribute to materials science and nanoscience/nanotechnology.

Polarization multiplexing for double images display

Metasurface provides subwavelength structures for manipulating wavefronts of light. The benefits of subwavelength components offer a continuous modulation of amplitude, phase, and polarization, thus eliminating the production of higher-order images and improving the utilization of light intensity. Despite the rapid progress in this field, multiparameter control of light using single layer metasurface is rarely reported. In fact, multiparameter control of light helps to improve information storage capacity and image fidelity. With simultaneous manipulation of polarization and amplitude at each pixel, it is possible to encode two separate images into one metasurface and reconstruct them under proper conditions. In a proof of concept experiment, we demonstrate an independent display of two binary images at the same position with polarization de-multiplexing from a single metasurface. This unique technology of encoding two images through amplitude and polarization manipulation provides a new opportunity for various applications in, such as encryption, information storage, polarization holograms, optical communications and fundamental physics.

Mechatronics Design, Modeling, and Characterization of a Soft Robotic Table for Object Manipulation on Surface

This paper presents the mechatronics design, modeling, and characterization of a soft robotic table inspired by caterpillar locomotion. The table is capable of manipulating objects on the XY-horizontal plane through surface deformations. The full realization of the table system including its actuation system, pneumatic system, electrical system, and user interface is discussed. The analytical modeling of these systems is explained. A design technique for the table is presented where its performance can be tuned by modifying its parameters via finite element analysis. The characteristics of the soft table are evaluated in terms of its hardware capabilities and object manipulation capabilities. These are important for the development of the table's control system. The key functional and operational parameters of the table such as the maximum object load, operational speed, and power consumptions are discussed. Experiments are conducted on three distinct objects to analyze the performance of object movement. Object transportability is analyzed based on the object's shape, size, and weight. The unique features of the table including the intrinsic soft handling of objects and simultaneous multiple object manipulation are validated. The soft table in action of various object manipulations can be viewed in the video supplied.

A vector holographic optical trap

The invention of optical tweezers almost forty years ago has triggered applications spanning multiple disciplines and has also found its way into commercial products. A major breakthrough came with the invention of holographic optical tweezers (HOTs), allowing simultaneous manipulation of many particles, traditionally done with arrays of scalar beams. Here we demonstrate a vector HOT with arrays of digitally controlled Higher-Order Poincaré Sphere (HOPS) beams. We employ a simple set-up using a spatial light modulator and show that each beam in the array can be manipulated independently and set to an arbitrary HOPS state, including replicating traditional scalar beam HOTs. We demonstrate trapping and tweezing with customized arrays of HOPS beams comprising scalar orbital angular momentum and cylindrical vector beams, including radially and azimuthally polarized beams simultaneously in the same trap. Our approach is general enough to be easily extended to arbitrary vector beams, could be implemented with fast refresh rates and will be of interest to the structured light and optical manipulation communities alike.

INFORMATIONAL AND ANALYTICAL SUPPLY OF PPP PROJECTS IMPLEMENTED BY REGIONAL CONSTRUCTION CLASS

A method of rationalizing the selection of a team for the communicative accompaniment of the PPP of projects implemented by the regional building cluster is proposed. The method is based on the application of a genetic algorithm, which allows simultaneous manipulation of many parameters and generate new through the use of different operators. Unlike other optimization methods, genetic algorithms tend to analyze different areas of the decision space simultaneously and therefore more suited to finding new domains with the best values of the target function. Solving the task of forming a team for the implementation of innovative construction projects does not require a strict finding of the global optimum on the investigated parameter space - in the shortest time to find the best of acceptable, local solution. The sequencing of the implementation of the genetic algorithm is described. The special effect of the algorithm is determined by the ability to search for through the survival of the strongest alternative solutions in uncertain fuzzy conditions. The procedure for finding the strongest alternative is described. Key words: construction company, public private partnership, genetic algorithm, innovation development, management tool, fuzzy conditions, uncertainty, strategic changes. кандидат технічних наук , Чуприна Ю . А . Інформаційно - аналітичний супровід ДПП проектів , що реалізуються регіональним будівельним кластером / Київський національний університет будівництва і архітектури , Україна , Київ Запропоновано метод раціоналізації підбору колективу для комунікативного супроводу ДПП проектів що реалізуються регіональним будівельним кластером. Метод ґрунтується на застосуванні генетичного алгоритму, що дозволяє одночасно маніпулювати багатьма параметрами і генерувати нові рішення за допомогою застосування різних операторів. На відміну від інших методів оптимізації генетичні алгоритми, як правило, аналізують різні області простору одночасно і тому більш пристосовані до знаходження нових областей з найкращими значеннями цільової функції. Розв’язання задачі формування колективу для виконання інноваційних будівельних проектів не вимагає строгого знаходження глобального оптимуму на досліджуваному просторі параметрів,- досить за короткий час знайти найкраще з прийнятних, локальне рішення. Описано послідовність виконання генетичного алгоритму. Особливий ефект алгоритму визначається здатністю здійснювати пошук за рахунок виживання найсильніших альтернативних рішень в невизначених нечітких умовах. Описана процедура пошуку найсильнішої альтернативи. Ключові слова: будівельне підприємство,державне приватне партнерство, генетичний алгоритм, інноваційний розвиток, інструмент управління, нечіткі умови, невизначеність, стратегічні зміни.

Gated Single-Molecule Transport in Double-Barreled Nanopores.

Single-molecule methods have been rapidly developing with the appealing prospect of transforming conventional ensemble-averaged analytical techniques. However, challenges remain especially in improving detection sensitivity and controlling molecular transport. In this article, we present a direct method for the fabrication of analytical sensors that combine the advantages of nanopores and field-effect transistors for simultaneous label-free single-molecule detection and manipulation. We show that these hybrid sensors have perfectly aligned nanopores and field-effect transistor components making it possible to detect molecular events with up to near 100% synchronization. Furthermore, we show that the transport across the nanopore can be voltage-gated to switch on/off translocations in real time. Finally, surface functionalization of the gate electrode can also be used to fine tune transport properties enabling more active control over the translocation velocity and capture rates.