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Nucleosome DNA unwrapping and its disassembly into hexasomes and tetrasomes is necessary for genomic access and plays an important role in transcription regulation. Previous single-molecule mechanical nucleosome unwrapping revealed a low- and a high-force transitions, and force-FRET pulling experiments showed that DNA unwrapping is asymmetric, occurring always first from one side before the other. However, the assignment of DNA segments involved in these transitions remains controversial. Here, using high-resolution optical tweezers with simultaneous single-molecule FRET detection, we show that the low-force transition corresponds to the undoing of the outer wrap of one side of the nucleosome (~27 bp), a process that can occur either cooperatively or noncooperatively, whereas the high-force transition corresponds to the simultaneous unwrapping of ~76 bp from both sides. This process may give rise stochastically to the disassembly of nucleosomes into hexasomes and tetrasomes whose unwrapping/rewrapping trajectories we establish. In contrast, nucleosome rewrapping does not exhibit asymmetry. To rationalize all previous nucleosome unwrapping experiments, it is necessary to invoke that mechanical unwrapping involves two nucleosome reorientations: one that contributes to the change in extension at the low-force transition and another that coincides but does not contribute to the high-force transition. [ABSTRACT FROM AUTHOR]

This paper presents the upconversion emission and magnetic properties of NaGdF 4 :Yb, Er nanoparticles separately doped with Fe and Mg in the 10–40% concentration. The XRD analysis validates the structural properties of NaGdF 4 :Yb, Er nanoparticles with different concentrations of the dopants. The XPS spectra of Fe-doped NaGdF 4 :Yb, Er and Mg-doped NaGdF 4 :Yb, Er nanoparticles reveal the presence of the dopants and their valency. The 30%Mg doping resulted in an intense red upconversion emission at 650 nm compared to the Fe-doped NaGdF 4 :Yb, Er due to the high lifetime of 254 μs for the 4F 9/2 → 4I 15/2 (660 nm) energy level transitions of Er3+ ion. The Mg-doped nanoparticles showed a 2.7 red-to-green emission intensity ratio, 5.7 for Fe doping at the same 30% concentration and above this doping level, upconversion emission quenching resulted. The room temperature VSM study demonstrated paramagnetic property in the Mg-doped NaGdF 4 :Yb, Er nanoparticles characterized by negligible coercivity and retentivity with low saturation magnetization. Similar behaviour is also observed for the Fe-doped nanoparticles up to 20% concentration, and it became weak ferromagnetic at a high 30 and 40% dopant. In addition, the present work demonstrated the optical trapping of individual magnetic nanoparticles of 30%Fe-doped NaGdF 4 :Yb, Er by using a single beam optical tweezer technique under the tightly focused 980 nm diode laser interaction and the estimated trap stiffnesses is 1.85 pN μm−1 W−1. The multifunctional upconversion red emission, magnetic and optical trapping properties obtained in NaGdF 4 :Yb,Er,30%Fe nanoparticles can be used as a single particle probe in the theragnostic application. [ABSTRACT FROM AUTHOR]

Modern medicine increases the demand for safe blood products. Ex vivo cultured red blood cells (cRBC) are eagerly awaited as a standardized, safe source of RBC. Established culture models still lack the terminal cytoskeletal remodeling from reticulocyte to erythrocyte with changes in the biomechanical properties and interacts with membrane stiffness, viscosity of the cytoplasm and the cytoskeletal network. Comprehensive data on the biomechanical properties of cRBC are needed to take the last step towards translation into clinical use in transfusion medicine. Aim of the study was the comparative analysis of topographical and biomechanical properties of cRBC, generated from human CD34+ adult hematopoietic stem/progenitor cells, with native reticulocytes (nRET) and erythrocytes (nRBC) using cell biological and biomechanical technologies. To gain the desired all-encompassing information, a single method was unsatisfactory and only the combination of different methods could lead to the goal. Topographical information was matched with biomechanical data from optical tweezers (OT), atomic force microscopy (AFM) and digital holographic microscopy (DHM). Underlying structures were investigated in detail. Imaging, deformability and recovery time showed a high similarity between cRBC and nRBC. Young's modulus and plasticity index also confirmed this similarity. No significant differences in membrane and cytoskeletal proteins were found, while lipid deficiency resulted in spherical, vesiculated cells with impaired biomechanical functionality. The combination of techniques has proven successful and experiments underscore a close relationship between lipid content, shape and biomechanical functionality of RBC. [ABSTRACT FROM AUTHOR]

One-dimensional (1D) target search is a well-characterized phenomenon for many DNA-binding proteins but is poorly understood for chromatin remodelers. Herein, we characterize the 1D scanning properties of SWR1, a conserved yeast chromatin remodeler that performs histone exchange on +1 nucleosomes adjacent to a nucleosome-depleted region (NDR) at gene promoters. We demonstrate that SWR1 has a kinetic binding preference for DNA of NDR length as opposed to gene-body linker length DNA. Using single and dual color single-particle tracking on DNA stretched with optical tweezers, we directly observe SWR1 diffusion on DNA. We found that various factors impact SWR1 scanning, including ATP which promotes diffusion through nucleotide binding rather than ATP hydrolysis. A DNA-binding subunit, Swc2, plays an important role in the overall diffusive behavior of the complex, as the subunit in isolation retains similar, although faster, scanning properties as the whole remodeler. ATP-bound SWR1 slides until it encounters a protein roadblock, of which we tested dCas9 and nucleosomes. The median diffusion coefficient, 0.024 μm²/s, in the regime of helical sliding, would mediate rapid encounter of NDR-flanking nucleosomes at length scales found in cellular chromatin. [ABSTRACT FROM AUTHOR]

At the single-cell level, cellular parameters, gene expression and cellular function are assayed on an individual but not population-average basis. Essential to observing and analyzing the heterogeneity and behavior of these cells/clusters is the ability to prepare and manipulate individuals. Here, we demonstrate a versatile microsystem, a stereo acoustic streaming tunnel, which is triggered by ultrahigh-frequency bulk acoustic waves and highly confined by a microchannel. We thoroughly analyze the generation and features of stereo acoustic streaming to develop a virtual tunnel for observation, pretreatment and analysis of cells for different single-cell applications. 3D reconstruction, dissociation of clusters, selective trapping/release, in situ analysis and pairing of single cells with barcode gel beads were demonstrated. To further verify the reliability and robustness of this technology in complex biosamples, the separation of circulating tumor cells from undiluted blood based on properties of both physics and immunity was achieved. With the rich selection of handling modes, the platform has the potential to be a full-process microsystem, from pretreatment to analysis, and used in numerous fields, such as in vitro diagnosis, high-throughput single-cell sequencing and drug development. [ABSTRACT FROM AUTHOR]

Compact modeling of charge trapping processes in GaN transistors is of fundamental importance for advanced circuit design. The goal of this article is to propose a methodology for modeling the dynamic characteristics of GaN power HEMTs in the realistic case where trapping/detrapping kinetics are described by stretched exponentials, contrary to ideal pure exponentials, thus significantly improving the state of the art. The analysis is based on: 1) an accurate methodology for describing stretched-exponential transients and extracting the related parameters and 2) a novel compact modeling approach, where the stretched exponential behavior is reproduced via multiple RC networks, whose parameters are specifically tuned based on the results of 1). The developed compact model is then used to simulate the transient performance of the HEMT devices as a function of duty cycle and frequency, thus providing insight on the impact of traps during the realistic switching operation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The quantum matter synthesizer (QMS) is a new quantum simulation platform in which individual particles in a lattice can be resolved and re-arranged into arbitrary patterns. The ability to spatially manipulate ultracold atoms and control their tunneling and interactions at the single-particle level allows full control of a many-body quantum system. We present the design and characterization of the QMS, which integrates into a single ultra-stable apparatus a two-dimensional optical lattice, a moving optical tweezer array formed by a digital micromirror device, and site-resolved atomic imaging. We demonstrate excellent mechanical stability between the lattice and tweezer array with relative fluctuations below 10 nm, diffraction-limited imaging at a resolution of 655 nm, and high-speed real-time control of the tweezer array at a 2.52 kHz refresh rate, which will be adopted to realize fast rearrangement of atoms. The QMS also features new technologies and schemes, such as nanotextured anti-reflective windows and all-optical long-distance transport of atoms. [ABSTRACT FROM AUTHOR]

Compared to conventional devices, metasurfaces offer the advantages of being lightweight, with planarization and tuning flexibility. This provides a new way to integrate and miniaturize optical systems. In this paper, a metasurface capable of generating multiple bottle beams was designed. Based on the Pancharatnamâ€"Berry (Pâ€"B) phase principle, the metasurface lens can accurately control the wavefront by adjusting the aspect ratio of the titanium dioxide nanopillars and the rotation angle. When irradiated by left-handed circularly polarized light with a wavelength of 632.8 nm, the optical system can produce multiple micron bottle beams. Taking two bottle beams as examples, the longitudinal full widths at half-maximum of the optical tweezers can reach 0.85 ÎĽm and 1.12 ÎĽm, respectively, and the transverse full widths at half-maximum can reach 0.46 ÎĽm and 0.6 ÎĽm. Also, the number of generated bottle beams can be varied by controlling the size of the annular obstacle. By changing the x -component of the unit rotation angle, the metasurface can also change the shape of the bottle beam â€" the beam cross-section can be changed from circular to elliptical. This paper also analyzes the trapping of ytterbium atoms by the multi bottle beam acting as optical tweezers. It is found that the multi bottle beam can cool and trap multiple ytterbium atoms. [ABSTRACT FROM AUTHOR]

Cellular and tissue biosystems emerge from the assembly of their constituent molecules and obtain a set of specific material properties. To measure these properties and understand how they influence cellular function is a central goal of mechanobiology. Froma bottoms-up, physics or engineering point-of-view, such systems are a composition of basic mechanical elements. However, the sheer number and dynamic complexity of them, including active molecular machines and their emergent properties, makes it currently intractable to calculate how biosystems respond to forces. Because many diseases result from an aberrant mechanotransduction, it is thus essential to measure this response. Recent advances in the technology of optical tweezers have broadened their scope from single-molecule applications to measurements inside complex cellular environments, even within tissues and animals. Here, we summarize the basic optical trapping principles, implementations and calibration procedures that enable force measurements using optical tweezers directly inside cells of living animals, in combination with complementary techniques. We review their versatility to manipulate subcellular organelles and measure cellular frequency-dependent mechanics in the piconewton force range from microseconds to hours. As an outlook, we address future challenges to fully unlock the potential of optical tweezers for mechanobiology. [ABSTRACT FROM AUTHOR]

Fusion of transmitter-containing vesicles with plasma membranes at the synaptic and neuromuscular junctions mediates neurotransmission and muscle contractions, respectively, thereby underlying all thoughts and actions. The fusion process is driven by the coupled folding and assembly of three synaptic SNARE proteins––syntaxin-1 and SNAP-25 on the target plasma membrane (t-SNAREs) and VAMP2 on the vesicular membrane (v-SNARE) into a four-helix bundle. Their assembly is chaperoned by Munc18-1 and many other proteins to achieve the speed and accuracy required for neurotransmission. However, the physiological pathway of SNARE assembly and its coupling to membrane fusion remains unclear. Here, we review recent progress in understanding SNARE assembly and membrane fusion, with a focus on results obtained by single-molecule manipulation approaches and electric recordings of single fusion pores. We describe two pathways of synaptic SNARE assembly, their associated intermediates, energetics, and kinetics. Assembly of the three SNAREs in vitro begins with the formation of a t-SNARE binary complex, on which VAMP2 folds in a stepwise zipper-like fashion. Munc18-1 significantly alters the SNARE assembly pathway: syntaxin-1 and VAMP2 first bind on the surface of Munc18-1 to form a template complex, with which SNAP-25 associates to conclude SNARE assembly and displace Munc18-1. During membrane fusion, multiple trans-SNARE complexes cooperate to open a dynamic fusion pore in a manner dependent upon their copy number and zippering states. Together, these results demonstrate that stepwise and cooperative SNARE assembly drive stagewise membrane fusion. [ABSTRACT FROM AUTHOR]

Dual-species single-atom array in optical tweezers has several advantages over the single-species atom array as a platform for quantum computing and quantum simulation. Thus, creating the defect-free dual-species single-atom array with atom numbers over hundreds is essential. As recent experiments demonstrated, one of the main difficulties lies in designing an efficient algorithm to rearrange the stochastically loaded dual-species atoms arrays into arbitrary demanded configurations. We propose a heuristic connectivity optimization algorithm to provide the near-fewest number of atom moves. Our algorithm introduces the concept of using articulation points in an undirected graph to optimize connectivity as a critical consideration for arranging the atom moving paths. Tested in array size of hundreds atoms and various configurations, our algorithm shows a high success rate (>97%), low extra atom moves ratio, good scalability, and flexibility. Furthermore, we propose a complementary step to solve the problem of atom loss during the rearrangement. [ABSTRACT FROM AUTHOR]

We discussed evolution characteristics of optical vortices with the cross phase in free space and found that both splitting and rotating can occur for vortices with circular symmetry due to cross phase. The rotation is bidirectional and the rotational direction will reverse at some certain distance. The physical mechanism behind the splitting and rotating of vortices is considered to be the effective anisotropic diffraction, which results from the cross phase. We also investigated the dynamics of superimposed pattern with two astigmatic vortices of opposite cross phase. The rotating of vortices always happens, however, the splitting only occurs for nonzero net cross phase coefficient. The results in the paper provide us with a theoretical method to control the dynamics of vortices by adjusting the cross phase, then may find potential applications in the field of optical tweezers. [ABSTRACT FROM AUTHOR]

• The importance of nuclease-free and active protein. • Gel-based assays show enzyme polarity and duplex unwinding activity. • Fluorescence-based helicase assays reveal DNA unwinding in real-time. • Continuous NTPase assays provide kinetic parameters for energy coupling. • Optical tweezers and fluorescence imaging enable visual biochemistry. • Magnetic tweezers provide single base-pair resolution. There are multiple assays available that can provide insight into the biochemical mechanism of DNA helicases. For the first 22 years since their discovery, bulk-phase assays were used. These include gel-based, spectrophotometric, and spectrofluorometric assays that revealed many facets of these enzymes. From 2001, single-molecule studies have contributed additional insight into these DNA nanomachines to reveal details on energy coupling, step size, processivity as well as unique aspects of individual enzyme behavior that were masked in the averaging inherent in ensemble studies. In this review, important aspects of the study of helicases are discussed including beginning with active, nuclease-free enzyme, followed by several bulk-phase approaches that have been developed and still find widespread use today. Finally, two single-molecule approaches are discussed, and the resulting findings are related to the results obtained in bulk-phase studies. [ABSTRACT FROM AUTHOR]

YBa 2 Cu 3 O 7−δ (YBCO) single grain, as a pivotal and popular oxide ceramic, is of great importance in practical applications for its considerable potential in magnetic properties. Normally, sizable YBCO bulk superconductors are fabricated by multi-seeded melt-growth for time saving. However, the performance of such bulks has long suffered from certain methodological barriers related to artificially induced imperfect-seeding, involving unreliable precision in seeding alignment and detrimental impact on grain boundaries. Here, a facile but effective in situ self-assembly (ISSA) strategy is reported for seeding structural design, in which exactly (110)/(110)-aligned twin-seeds naturally arise through the self-replication of crystallographic orientation from one small seed during thermal procedure. Distinctively, after two induced grains impinging, a [110]-oriented growth zone emerges and expands to the edge of the sample, leading to a contamination-free homo-combination. As a result, 25 mm diameter fully-grown bulks with single-grain quality were reliably attained, which exhibited superior levitation forces up to 65.1 N and field trapping capability up to 0.8943 T with a prominent feature of mono-peak. The ISSA seeding strategy in this work, with its nature in achieving exact seeding alignment, is widely applicable for the preparation of other high-performance bulks in the YBCO family. [ABSTRACT FROM AUTHOR]

Recently, optical coordinate transformation has garnered considerable research interest for manipulating structured light in emerging optical communication applications. Herein, we propose a circle-Cassinian optical coordinate transformation based on polarization invariance to generate an elliptical-symmetry radially polarized (ESRP) optical beam. Accordingly, we designed three isotropic phase plates for placement in the 4f optical system. The numerical simulations demonstrated accurate generation of the ESRP beam with specified intensity, including an overall intensity adjustment applied in the input plane. Therefore, the proposed method can aid in designing vector light fields, and the ESRP beam can be applied to optical tweezers and surface plasmonic-field generation. [ABSTRACT FROM AUTHOR]

The quantitative analysis of cell surface antigens has attracted increasing attention due to the antigenic variation recognition that can facilitate early diagnoses. This paper presents a novel methodology based on the optical "cell-tearing" and the especially proposed "dilution regulations" to detect variations in cell surface antigens. The cell attaches to the corresponding antibody-coated slide surface. Then, the cell-binding firmness between a single cell and the functionalized surface is assayed by optically tearing using gradually reduced laser powers incorporated with serial antibody dilutions. Groups B and B3 of red blood cells (RBCs) were selected as the experiment subject. The results indicate that a higher dilution called for lower power to tear off the cell binding. According to the proposed relative-quantitative analysis theory, antigenic variation can be intuitively estimated by comparing the maximum allowable dilution folds. The estimation result shows good consistency with the finding in the literature. This study suggests a novel methodology for examining the variation in cell surface antigens, expected to be widely capable with potential sensor applications not only in biochemistry and biophysics, but also in the micro-/nano- engineering field. [ABSTRACT FROM AUTHOR]

Synaptotagmin-1 (Syt1) is a vesicular calcium sensor required for synchronous neurotransmitter release, composed of a single-pass transmembrane domain linked to two C2 domains (C2A and C2B) that bind calcium, acidic lipids, and SNARE proteins that drive fusion of the synaptic vesicle with the plasma membrane. Despite its essential role, how Syt1 couples calcium entry to synchronous release is poorly understood. Calcium binding to C2B is critical for synchronous release, and C2B additionally binds the SNARE complex. The C2A domain is also required for Syt1 function, but it is not clear why. Here, we asked what critical feature of C2A may be responsible for its functional role and compared this to the analogous feature in C2B. We focused on highly conserved poly-lysine patches located on the sides of C2A (K189-192) and C2B (K324-327). We tested effects of charge-neutralization mutations in either region (Syt1K189-192A and Syt1K326-327A) side by side to determine their relative contributions to Syt1 function in cultured cortical neurons from mice of either sex and in single-molecule experiments. Combining electrophysiological recordings and optical tweezers measurements to probe dynamic single C2 domain–membrane interactions, we show that both C2A and C2B polybasic patches contribute to membrane binding, and both are required for evoked release. The size of the readily releasable vesicle pool and the rate of spontaneous release were unaffected, so both patches are likely required specifically for synchronization of release. We suggest these patches contribute to cooperative membrane binding, increasing the overall affinity of Syt1 for negatively charged membranes and facilitating evoked release. [ABSTRACT FROM AUTHOR]

The endosomal sorting complexes required for transport (ESCRT) system is an ancient and ubiquitous membrane scission machinery that catalyzes the budding and scission of membranes. ESCRT-mediated scission events, exemplified by those involved in the budding of HIV-1, are usually directed away from the cytosol ("reverse topology"), but they can also be directed toward the cytosol ("normal topology"). The ESCRT-III subunits CHMP1B and IST1 can coat and constrict positively curved membrane tubes, suggesting that these subunits could catalyze normal topology membrane severing. CHMP1B and IST1 bind and recruit the microtubule-severing AAA+ ATPase spastin, a close relative of VPS4, suggesting that spastin could have a VPS4-like role in normaltopology membrane scission. Here, we reconstituted the process in vitro using membrane nanotubes pulled from giant unilamellar vesicles using an optical trap in order to determine whether CHMP1B and IST1 are capable of membrane severing on their own or in concert with VPS4 or spastin. CHMP1B and IST1 copolymerize on membrane nanotubes, forming stable scaffolds that constrict the tubes, but do not, on their own, lead to scission. However, CHMP1B-IST1 scaffolded tubes were severed when an additional extensional force was applied, consistent with a friction-driven scission mechanism. We found that spastin colocalized with CHMP1B-enriched sites but did not disassemble the CHMP1B-IST1 coat from the membrane. VPS4 resolubilized CHMP1B and IST1 without leading to scission. These observations show that the CHMP1B-IST1 ESCRT-III combination is capable of severing membranes by a friction-driven mechanism that is independent of VPS4 and spastin. [ABSTRACT FROM AUTHOR]

Cells are known to continuously remodel their local extracellular matrix (ECM) and in a reciprocal way, they can also respond to mechanical and biochemical properties of their fibrous environment. In this study, we measured how stiffness around dermal fibroblasts (DFs) and human fibrosarcoma HT1080 cells differs with concentration of rat tail type 1 collagen (T1C) and type of ECM. Peri-cellular stiffness was probed in four directions using multi-axes optical tweezers active microrheology (AMR). First, we found that neither cell type significantly altered local stiffness landscape at different concentrations of T1C. Next, rat tail T1C, bovine skin T1C and fibrin cell-free hydrogels were polymerized at concentrations formulated to match median stiffness value. Each of these hydrogels exhibited distinct fiber architecture. Stiffness landscape and fibronectin secretion, but not nuclear/cytoplasmic YAP ratio differed with ECM type. Further, cell response to Y27632 or BB94 treatments, inhibiting cell contractility and activity of matrix metalloproteinases, respectively, was also dependent on ECM type. Given differential effect of tested ECMs on peri-cellular stiffness landscape, treatment effect and cell properties, this study underscores the need for peri-cellular and not bulk stiffness measurements in studies on cellular mechanotransduction. [ABSTRACT FROM AUTHOR]

Acid-sensing ion channels (ASICs) are neuronal, proton-gated, Na+-selective ion channels. They are involved in various physiological and pathological processes such as neurodegeneration after stroke, pain sensation, fear behavior and learning. To obtain information on the activation mechanism of ASIC1a, we attempted in this study to impose distance constraints between paired residues in different channel domains by using cross-linkers reacting with engineered Cys residues, and we measured how this affected channel function. First, the optical tweezer 4′-Bis(maleimido)azobenzene (BMA) was used, whose conformation changes depending on the wavelength of applied light. After exposure of channel mutants to BMA, an activation of the channel by light was only observed with a mutant containing a Cys mutation in the extracellular pore entry, I428C. Western blot analysis indicated that BMA did not cross-link Cys428 residues. Extracellular application of methanethiosulfonate (MTS) cross-linkers of different lengths changed the properties of several Cys mutants, in many cases likely without cross-linking two Cys residues. Our observations suggest that intersubunit cross-linking occurred in the wrist mutant A425C and intrasubunit cross-linking in the acidic pocket mutant D237C/I312C. In these mutants, exposure to cross-linkers favored a non-conducting channel conformation and induced an acidic shift of the pH dependence and a decrease of the maximal current amplitude. Overall, the cross-linking approaches appeared to be inefficient, possibly due to the geometrical requirements for successful reactions of the two ends of the cross-linking compound. [ABSTRACT FROM AUTHOR]

There are numerous ways to control objects in the Stokes regime, with microscale examples ranging from the use of optical tweezers to the application of external magnetic fields. In contrast, there are relatively few explorations of theoretical controllability, which investigate whether or not refined and precise control is indeed possible in a given system. In this work, seeking to highlight the utility and broad applicability of such rigorous analysis, we recount and illustrate key concepts of geometric control theory in the context of multiple particles in Stokesian fluids interacting with each other, such that they may be readily and widely applied in this largely unexplored fluid-dynamical setting. Motivated both by experimental and abstract questions of control, we exemplify these techniques by explicit and detailed application to multiple problems concerning the control of two particles, such as the motion of tracers in flow and the guidance of one sphere by another. Further, we showcase how this analysis of controllability can directly lead to the construction of schemes for control, in addition to facilitating explorations of mechanical efficiency and contributing to our overall understanding of non-local hydrodynamic interactions in the Stokes limit. [ABSTRACT FROM AUTHOR]

Gaining control on particle–particle interactions and in this way on their (self)‐assembled structures is essentialfor colloidal and material sciences. Currently, different strategies are described to achieve such control, however, all of them lack the spatiotemporal resolution required at the microscale. In this work, the potential of combining optical trapping and resonant photoexcitation for modifying particle–particle interactions and subsequent assembling of dye‐doped particles at the solution interface is demonstrated. The particle assemblies prepared by nonresonant 1064 nm optical trapping undergo morphology changes after resonant photoexcitation of the embedded dye molecules. Depending on the physicochemical properties of interface, quick hexagonal close packing (HCP)‐rearrangement or explosive dispersion of assemblies is observed at air/solution (A/S) and glass/solution interfaces, respectively. By contrast, by resonant photoexcitation only, the dispersed dye‐doped particles are pushed toward the A/S interface, followed by association to yield HCP‐structured assemblies. The results are rationalized by considering the optical absorption force coupled with other nonoptical forces (e.g., capillary force, dipole–dipole or electrostatic repulsion) at the solution interface. Due to the inherent spatiotemporal properties of light and electronic transition of materials, absorption force is a unique element to control and modify the structural order of particle assemblies at interfaces. [ABSTRACT FROM AUTHOR]

The elastic properties of the double-stranded DNA handles used in optical tweezers experiments on biomolecules are customarily modeled by an extensible worm-like chain model. Fitting such a model to experimental data, however, is no trivial task, as the function depends on four parameters in a highly non-linear fashion. We hereby propose a method to bypass the fitting procedure and obtain an empirical force vs. extension curve that accurately reproduces the elasticity of the handles. [ABSTRACT FROM AUTHOR]

Online manipulation of multiple micro- and nanoscale agents is of major interest for various research applications. Among the biggest limitations of wireless external actuation are its global and coupled influences in the workspace, which limit the robust manipulation of multiple agents independently and simultaneously. In this paper, we propose novel motion planning algorithms, $\tt Bi$ - $\tt iSST$ and $\tt Ref$ - $\tt iSST$ , to quickly generate time-optimal trajectories for multiple agents sharing global external fields. Both algorithms are extended by the stable sparse rapidly-exploring random tree kinodynamic motion planning algorithm. The $\tt Bi$ - $\tt iSST$ uses a bidirectional approach to speed up the searching process. A novel connection process is proposed to connect the two trees efficiently by applying an optimization procedure. The $\tt Ref$ - $\tt iSST$ uses the workspace information to quickly generate global-routing trajectories as references, then guides the search process more effectively by getting more accurate heuristics according to the reference global-routing trajectories. A transition matrix similar to that in Markov Decision Processes is used to form the reference trajectory. Compared with the state-of-the-art $\tt iSST$ algorithm, the proposed algorithms quickly update feasible solutions and converge to a near-optimal, minimum-time solution to increase the efficiency of the simultaneous manipulation of multiple micro agents using global external fields. Extensive analysis and physical experiments are presented to confirm the effectiveness and the performance of the motion planning algorithms. Note to Practitioners—Autonomous manipulation of multiple micro- and nanoscale agents is of major interest for various research applications. Wireless actuation is a promising way to position those objects. The commonly used non-contact actuation techniques include magnetic actuation, electrical field actuation, optical tweezers, and actuated flows, etc. Among the biggest limitations of wireless external actuation are its global and coupled influences in the workspace, which limit the capability to robustly manipulate multiple agents independently and simultaneously. In this paper, we propose novel bidirectional informed sampling-based motion planning algorithms to quickly generate time-optimal trajectories for multiple agents sharing global external fields. Novel heuristics and informed reference trajectory are used to guide the search for manipulating multiple agents under global fields. Numerical simulations and physical experiments are presented to demonstrate the performance of the motion planning design. The proposed algorithm guarantees anytime performance and quickly converges to a near-optimal minimum time solution for multiple agents. Although we focus on the electric-field actuated multiple-micro-agent system, the proposed motion planning strategies are not limited to the actuation and can be generalized to other field-based applications, in which the actuation among a group of multiple agents are coupled or intertwined. [ABSTRACT FROM AUTHOR]

Single‐cell RNA sequencing (scRNA‐seq) is a powerful technology for revealing the heterogeneity of cellular states. However, existing scRNA‐seq platforms that utilize bead‐based technologies suffer from a large number of empty microreactors and a low cell/bead capture efficiency. Here, Well‐paired‐seq is presented, which consists of thousands of size exclusion and quasi‐static hydrodynamic dual wells to address these limitations. The size‐exclusion principle allows one cell and one bead to be trapped in the bottom well (cell‐capture‐well) and the top well (bead‐capture‐well), respectively, while the quasi‐static hydrodynamic principle ensures that the trapped cells are difficult to escape from cell‐capture‐wells, achieving cumulative capture of cells and effective buffer exchange. By the integration of quasi‐static hydrodynamic and size‐exclusion principles, the dual wells ensure single cells/beads pairing with high density, achieving excellent efficiency of cell capture (≈91%), cell/bead pairing (≈82%), and cell‐free RNA removal. The high utilization of microreactors and single cells/beads enable to achieve a high throughput (≈105 cells) with low collision rates. The technical performance of Well‐paired‐seq is demonstrated by collecting transcriptome data from around 200 000 cells across 21 samples, successfully revealing the heterogeneity of single cells and showing the wide applicability of Well‐paired‐seq for basic and clinical research. [ABSTRACT FROM AUTHOR]

It is well known that positions of topological defects (TDs) in liquid crystals can be manipulated experimentally by locally distorting the liquid crystalline (LC) order, as for example by melting induced by optical tweezers. In this work, we study numerically the nematic ordering profiles and the corresponding topological defect configurations in thin nematic liquid crystalline shells controlled by imposed local distortion of LC order. We demonstrate that within curved LC films such manipulations could be strongly affected by local Gaussian curvature if it exhibits strong spatial variations. We use mesoscopic approach in which the shell geometry and LC orientational order are described by curvature of the surface and nematic order parameter tensor. For illustration purposes, we consider LC shells exhibiting spherical topology. We show that on increasing prolateness of shells, which imposes spatially inhomogeneous Gaussian curvature, TDs are relatively strongly "glued" to a local Gaussian curvature. [ABSTRACT FROM AUTHOR]

We demonstrate photophoretic force-based optical trapping of multiple absorbing particles in air by loosely focusing a Gaussian beam emanating from a single mode fiber using convex lenses of different focal lengths, and investigate the dependence of the number of trapped particles and their sizes on the focal length. We observe the formation of particle chains at a particular focal length, and measure the axial dynamic range of optical trapping for each lens system. We then develop a numerical simulation to explain this observed dynamic range by estimating the temperature distribution across a particle surface, and determining the axial photophoretic force. Our simulation results are in reasonable agreement with experimental results. Interestingly, we also observe that the average size of trapped particles reduces as we increase the lens focal lengths. This is somewhat intriguing as each lens produces the same intensity profile, albeit at different axial distances. However, the axial intensity gradient reduces as the lens focal length is increased, which suggests that such gradients may somehow be involved in the mechanism of photophoretic confinement. [ABSTRACT FROM AUTHOR]

Interactions between stromal and lymphoma cells in the bone marrow are closely related to drug resistance and therapy failure. Physiologically relevant pre-clinical three-dimensional (3D) models recapitulating lymphoma microenvironmental complexity do not currently exist. In this study, we proposed a scheme for optically controlled hybrid lymphoma spheroid formation with the use of optical tweezers (OT). Following the preparation of stromal spheroids using agarose hydrogel, two aggressive non-Hodgkin lymphoma B-cell lines, Ri-1 (DLBCL) and Raji (Burkitt lymphoma), were used to conduct multi-cellular spheroid formation driven by in-house-developed fluorescence optical tweezers. Importantly, the newly formed hybrid spheroid preserved the 3D architecture for the next 24 h. Our model was successfully used for the evaluation of the influence of the anticancer agents doxorubicin (DOX), ibrutinib (IBR), and AMD3100 (plerixafor) on the adhesive properties of lymphoma cells. Importantly, our study revealed that a co-treatment of DOX and IBR with AMD3100 affects the adhesion of B-NHL lymphoma cells. [ABSTRACT FROM AUTHOR]

Spatial light modulators (SLMs) are versatile devices used for optical studies. These instruments have a wide area of application in photonics. Additionally, SLMs have potential utility in different applications, such as biomedical applications, laser based surgery for precise cutting and as optical tweezers to separate cells in a petri container. However, the high cost of SLM devices prevents their widespread use in many areas, including industrial areas and scientific research laboratories. This paper demonstrates how to design a digital light processor (DLP) based low-cost SLM and describes how to obtain structured electromagnetic waves with the designed SLM. Therefore, this research was undertaken to design and produce a low-cost SLM device for optical applications. For this purpose, two prerequisites had to be fulfilled, the first was to use suitable components of a projection device with DLP-based digital micro-mirror device (DMD), and the second was to eliminate unnecessary SLM components from the system. Finally, holographic images reflected on the SLM screen were created by using Mathematica software program to change the amplitude and phase of the electromagnetic waves in order to obtain the structured electromagnetic waves. [ABSTRACT FROM AUTHOR]

The optothermal manipulation of micro-objects is significant for understanding and exploring the unknown in the microscale word, which has found many applications in colloidal science and life science. In this work, we study the transverse forces of an optothermal trap in front of a gold film, which is an absorbing reflective surface for the incident laser beam. It is demonstrated that optothermal forces can be divided into two parts: optical force of a standing-wave trap, and thermal force of a thermal trap. The optical force of the standing-wave trap can be obtained by measuring the optical trapping force close to a non-absorbing film with same reflectance. The thermal force can be obtained by subtracting the optical force of the standing-wave trap from the total trapping force of the optothermal trap close to the gold film. The results show that both optical and thermal trapping forces increase with laser power increasing. The optical trapping force is larger than the thermal trapping force, which is composed of convective drag force and thermophoretic force. Further experiment is run to study the composition of thermal force. The result shows that the convective flow is generated later than the thermophoretic flow. The results proposed here are useful for enabling users to optimize optothermal manipulation method for future applications. [ABSTRACT FROM AUTHOR]

In order to accelerate ions by a laser, we go back to the original and the fundamental idea of how longitudinal field structure generation can be carried out in an ionized media and how particles may be trapped by the created wakefield. The latter condition is characterized by the phase velocity of the longitudinal structure vph be equal to the particle trapping width vtr. Since the trapping width is inversely proportional to the square-root of the mass of the accelerated particles, this width is much shorter for ions than for electrons. Thus, our dictum for laser ion acceleration is to impose a near critical density trap to decelerate laser group velocity, vg and subsequently to generate longitudinal wakefield to be able to trap ions under the condition of vtr = vph. We demonstrate this concept by PIC simulation and find that this method is effective, and the efficiency of laser ion acceleration is enhanced by a couple of orders of magnitude toward unity. [ABSTRACT FROM AUTHOR]

We report a new method for the rapid identification of pathogenic bacterial species at the single‐cell level that combines laser tweezers Raman spectroscopy (LTRS) with deep learning (DL). LTRS can accurately measure single‐cell Raman spectra (scRS) without destroying and labeling cells. Based on the scRS data, DL rapidly and accurately identifies pathogenic bacteria. We measured scRS of 15 species bacteria using homemade LTRS. For each species, approximately, 160 cells from three different patients were measured, one patient's data were used as test set, and the rest after being augmented was used as training set. A residual network (ResNet) model, trained on the augmented training set, achieved an accuracy of 94.53% on the test set. Moreover, we applied gradient‐weighted class activation mapping to visualize the proposed model. Finally, we demonstrated the advantages of ResNet over traditional machine‐learning algorithms. [ABSTRACT FROM AUTHOR]

We researched the mechanical unfolding of protein domains in monomeric protein NuG2 and the tandem polyproteins (NuG2)8 and (NuG2)16 using a dual-trap optical tweezers system. By stretching NuG2 and its polyproteins, (NuG2)8 and (NuG2)16 at the constant pulling speed of 500 nm sâˆ'1, we achieved the mechanical unfolding force of each domain in these proteins. Besides, we calculated the energy dissipation of NuG2, (NuG2)8 and (NuG2)16 by measuring the area enclosed by stretching and relaxation traces. Our results represent a key step towards engineering artificial polyproteins with controllable mechanical force and energy dissipation properties for force-buffering and energy dissipator applications. [ABSTRACT FROM AUTHOR]

Originally described in crane-fly spermatocytes, tethers physically link and transmit force between the ends of separating chromosomes. Optical tweezers and laser scissors were used to sever the tether between chromosomes, create chromosome fragments attached to the tether which move toward the opposite pole, and to trap the tethered fragments. Laser microsurgery in the intracellular space between separating telomeres reduced chromosome strain in half of tested chromosome pairs. When the telomere-containing region was severed from the rest of the chromosome body, the resultant fragment either traveled towards the proper pole (poleward), towards the sister pole (cross-polar), or movement ceased. Fragment travel towards the sister pole varied in distance and always ceased following a cut between telomeres, indicating the tether is responsible for transferring a cross-polar force to the fragment. Optical trapping of cross-polar traveling fragments places an upper boundary on the tethering force of ~1.5 pN.

A semi-analytical model is proposed to describe the force generated by a near-field optical trap. The model contains fitting parameters that can be adjusted to resemble a reference force-field. The model parameters for a plasmonic near-field trap consisting of a C-shaped engraving are determined using least squares regression. The reference values required for the regression analysis are calculated using the Maxwell stress tensor method. The speed and accuracy of the proposed model are compared with the conventional method. The model is found to be significantly faster with an acceptable level of accuracy. [ABSTRACT FROM AUTHOR]

Background: Dengue virus (DENV) causes the most significant mosquito-borne viral disease with a wide spectrum of clinical manifestation, including neurological symptoms associated with lethal dengue diseases. Dopamine receptors are expressed in central nervous system, and dopamine antagonists have been reported to exhibit antiviral activity against DENV infection in vivo and in vitro. Although identification of host-cell receptor is critical to understand dengue neuropathogenesis and neurotropism, the involvement of dopamine receptors in DENV infection remains unclear., Results: We exploited the sensitivity and precision of force spectroscopy to address whether dopamine type-2 receptors (D2R) directly interact with DENV particles at the first step of infection. Using optical tweezers, we quantified and characterized DENV binding to D2R expressed on Chinese hamster ovary (CHO) cells. Our finding suggested that the binding was D2R- and DENV-dependent, and that the binding force was in the range of 50-60 pN. We showed that dopamine antagonists prochlorperazine (PCZ) and trifluoperazine (TFP), previously reported to inhibit dengue infection, interrupt the DENV-D2R specific binding., Conclusions: This study demonstrates that D2R could specifically recognize DENV particles and function as an attachment factor on cell surfaces for DENV. We propose D2R as a host receptor for DENV and as a potential therapeutic target for anti-DENV drugs., (© 2024. The Author(s).)

In recent years, Raman spectroscopy has garnered growing interest in the field of biomedical research. It offers a non-invasive and label-free approach to defining the molecular fingerprint of immune cells. We utilized Raman spectroscopy on optically trapped immune cells to investigate their molecular compositions. While numerous immune cell types have been studied in the past, the characterization of living human CD3/CD28-stimulated T cell subsets remains incomplete. In this study, we demonstrate the capability of Raman spectroscopy to readily distinguish between naïve and stimulated CD4 and CD8 cells. Additionally, we compared these cells with monocytes and discovered remarkable similarities between stimulated T cells and monocytes. This paper contributes to expanding our knowledge of Raman spectroscopy of immune cells and serves as a launching point for future clinical applications.

Mutation in oncogene KRas plays a crucial role in the occurrence and progression of numerous malignant tumors. Malignancy involves changes in cell mechanics for extensive cellular deformation during metastatic dissemination. We hypothesize that oncogene KRas mutations are intrinsic to alterations in cellular mechanics that promote malignant tumor generation and progression. Here, we demonstrate the use of optical tweezers coupled with a confocal fluorescence imaging system and gene interference technique to reveal that the mutant KRas protein can be transported between homogeneous and heterogeneous tumor cells by tunneling nanotubes (TNTs), resulting in a significant reduction of membrane tension and acceleration of membrane phospholipid flow in the recipient cells. Simultaneously, the changes in membrane mechanical properties of the tumor cells also enhance the metastatic and invasive ability of the tumors, which further contribute to the deterioration of the tumors. This finding helps to clarify the association between oncogene mutations and changes in the mechanical properties of tumor cells, which provides a theoretical basis for the development of cancer treatment strategies. STATEMENT OF SIGNIFICANCE: Here, we present a laser confocal fluorescence system integrated with optical tweezers to observe the transfer of mutant KRas G12D protein from mutant cells to wild-type cells through TNTs. Malignancy involves changes in cell mechanics for extensive cellular deformation during metastatic dissemination. Our results demonstrate a significant decrease in membrane tension and an increase in membrane phospholipid flow in recipient cells. These alterations in mechanical properties augment the migration and invasive capabilities of tumor cells, contributing to tumor malignancy. Our findings propose that cellular mechanical properties could serve as new markers for tumor development, and targeting membrane tension may hold potential as a therapeutic strategy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Many neurodegenerative diseases feature misfolded proteins that propagate via templated conversion of natively folded molecules. However, crucial questions about how such prion-like conversion occurs and what drives it remain unsolved, partly because technical challenges have prevented direct observation of conversion for any protein. We observed prion-like conversion in single molecules of superoxide dismutase-1 (SOD1), whose misfolding is linked to amyotrophic lateral sclerosis. Tethering pathogenic misfolded SOD1 mutants to wild-type molecules held in optical tweezers, we found that the mutants vastly increased misfolding of the wild-type molecule, inducing multiple misfolded isoforms. Crucially, the pattern of misfolding was the same in the mutant and converted wild-type domains and varied when the misfolded mutant was changed, reflecting the templating effect expected for prion-like conversion. Ensemble measurements showed decreased enzymatic activity in tethered heterodimers as conversion progressed, mirroring the single-molecule results. Antibodies sensitive to disease-specific epitopes bound to the converted protein, implying that conversion produced disease-relevant misfolded conformers., (© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Toehold-mediated strand displacement (TMSD) is extensively utilized in dynamic DNA nanotechnology and for a wide range of DNA or RNA-based reaction circuits. Investigation of TMSD kinetics typically relies on bulk fluorescence measurements providing effective, bulk-averaged reaction rates. Information on individual molecules or even base pairs is scarce. In this work, we explore the dynamics of strand displacement processes at the single-molecule level using single-molecule force spectroscopy with a microfluidics-enhanced optical trap supported by state-of-the-art coarse-grained simulations. By applying force, we can trigger and observe TMSD in real-time with microsecond and nanometer resolution. We find TMSD proceeds very rapidly under load with single step times of 1 µs. Tuning invasion efficiency by introducing mismatches allows studying thousands of forward/backward invasion events on a single molecule and analyze the kinetics of the invasion process. Extrapolation to zero force reveals single step times for DNA invading DNA four times faster than for RNA invading RNA. We also study the kinetics of DNA invading RNA, a process that in the absence of force would rarely occur. Our results reveal the importance of sequence effects for the TMSD process and have relevance for a wide range of applications in nucleic acid nanotechnology and synthetic biology., (© 2024. The Author(s).)

Optical trapping is a potent tool for achieving precise and noninvasive manipulation of small objects in a vacuum and liquids. However, due to the substantial disparity between optical forces and interfacial adhesion, target objects should be suspended in fluid environments, rendering solid contact surfaces a restricted area for conventional optical tweezers. In this work, by relying on a single continuous wave (CW) laser, we demonstrate an optical manipulation system applicable for both fluid and solid domains, namely, amphibious hybrid laser tweezers. The key to our system lies in modulating the intensity of the CW laser with duration shorter than the dynamic thermal equilibrium time within objects, wherein strong thermal gradient forces with ∼6 orders of magnitude higher than the forces in optical tweezers are produced, enabling moving and trapping micro/nano-objects on solid interfaces. Thereby, CW laser-based optical tweezers and pulsed laser-based photothermal shock tweezers are seamlessly fused with the advantages of cost-effectiveness and simplicity. Our concept breaks the stereotype that CW lasers are limited to generating tiny forces and instead achieve ultrawide force generation spanning from femto-newtons (10 -15 N) to (10 -6 N). Our work expands the horizon of optical manipulation by seamlessly bridging its applications in fluid and solid environments and holds promise for inspiring optical manipulation techniques to perform more challenging tasks, which may unearth application scenarios in diverse fields such as fundamental physical research, nanofabrication, micro/nanorobotics, biomedicine, and cytology.

In recent years, optical tweezers have become an effective bioassay tool due to their unique advantages, especially in combination with suspension beads, which can be applied to develop a high-performance analysis platform capable of high-quality imaging and stable signal output. However, the optical tweezer-assisted bead analysis is still at the early stage, and further development of different favorable methods is in need. Herein, we have first developed the optical tweezer-assisted immuno-rolling circle amplification (immuno-RCA) on beads for protein detection. Prostate-specific antigen was selected as the model analyte, and the immunosandwich structure on beads was built by the high affinity of "antibody-antigen". The "protein-nucleic acid" signals were effectively converted through the covalent coupling procedure of antibodies and oligonucleotides, further initiating the RCA reaction to achieve signal amplification. The individual beads with the strong irregular Brownian motion in a fluid environment were eventually trapped by the optical tweezers to acquire the accurate and high-quality signal. Compared with the conventional immunoassay on beads, the sensitivity of the developed strategy was increased by 587 times with a limit of detection of 4.29 pg/mL (0.13 pM), as well as excellent specificity, stability, and reproducibility. This study developed the new optical tweezer-assisted beads imaging strategy for protein targets, which has great potential for being applied to clinical serology research and expands the application of optical tweezers in the bioassays.

αβ T cell receptors (TCRs) principally recognize aberrant peptides bound to major histocompatibility complex molecules (pMHCs) on unhealthy cells, amplifying specificity and sensitivity through physical load placed on the TCR-pMHC bond during immunosurveillance. To understand this mechanobiology, TCRs stimulated by abundantly and sparsely arrayed epitopes (NP 366-374 /D b and PA 224-233 /D b , respectively) following in vivo influenza A virus infection were studied with optical tweezers. While certain NP repertoire CD8 T lymphocytes require many ligands for activation, others are digital, needing just few. Conversely, all PA TCRs perform digitally, exhibiting pronounced bond lifetime increases through sustained, energizing volleys of structural transitioning. Optimal digital performance is superior in vivo, correlating with ERK phosphorylation, CD3 loss, and activation marker up-regulation in vitro. Given neoantigen array paucity, digital TCRs are likely critical for immunotherapies.

Micro and nano-scale manipulation of living matter is crucial in biomedical applications for diagnostics and pharmaceuticals, facilitating disease study, drug assessment, and biomarker identification. Despite advancements, trapping biological nanoparticles remains challenging. Nanotweezer-based strategies, including dielectric and plasmonic configurations, show promise due to their efficiency and stability, minimizing damage without direct contact. Our study uniquely proposes an inverted hybrid dielectric-plasmonic nanobowtie designed to overcome the primary limitations of existing dielectric-plasmonic systems, such as high costs and manufacturing complexity. This novel configuration offers significant advantages for the stable and long-term trapping of biological objects, including strong energy confinement with reduced thermal effects. The metal's efficient light reflection capability results in a significant increase in energy field confinement (EC) within the trapping site, achieving an enhancement of over 90% compared to the value obtained with the dielectric nanobowtie. Numerical simulations confirm the successful trapping of 100 nm viruses, demonstrating a trapping stability greater than 10 and a stiffness of 2.203 fN/nm. This configuration ensures optical forces of approximately 2.96 fN with an input power density of 10 mW/μm 2 while preserving the temperature, chemical-biological properties, and shape of the biological sample.

We investigated the single particle kinetics of the molecular release processes from two types of microcapsules used as drug delivery systems (DDS): biodegradable poly(lactic- co -glycolic) acid (PLGA) and a light-triggered-degradable liposome encapsulating gold nanospheres (liposome-GNP). To optimize the design of DDS capsules, it is highly desirable to develop a method for real-time monitoring of the release process. Using a combination of optical tweezers and confocal fluorescence microspectroscopy we successfully analyzed a single optically trapped PLGA particle and liposome-GNPs in solution. From temporal decay profiles of the fluorescence intensity, we determined the time constant τ of the release processes. We demonstrated that the release rate of spontaneously degradable microcapsules (PLGA) decreased with increasing size, while conversely, the release rate of external stimuli-degradable microcapsules (liposome-GNPs) increased in proportion to their size. This result is explained by the differences in the disruption mechanisms of the capsules, with PLGA undergoing hydrolysis and the GNPs in the liposome-GNP undergoing a photoacoustic effect under nanosecond pulsed laser irradiation. The present approach offers a way forward to an alternative microanalysis system for single drug delivery nanocarriers.

TMPyP is a porphyrin capable of DNA binding and used in photodynamic therapy and G-quadruplex stabilization. Despite its broad applications, TMPyP's effect on DNA nanomechanics is unknown. Here we investigated, by manipulating λ-phage DNA with optical tweezers combined with microfluidics in equilibrium and perturbation kinetic experiments, how TMPyP influences DNA nanomechanics across wide ranges of TMPyP concentration (5-5120 nM), mechanical force (0-100 pN), NaCl concentration (0.01-1 M) and pulling rate (0.2-20 μm/s). Complex responses were recorded, for the analysis of which we introduced a simple mathematical model. TMPyP binding, which is a highly dynamic process, leads to dsDNA lengthening and softening. dsDNA stability increased at low (<10 nM) TMPyP concentrations, then decreased progressively upon increasing TMPyP concentration. Overstretch cooperativity decreased, due most likely to mechanical roadblocks of ssDNA-bound TMPyP. TMPyP binding increased ssDNA's contour length. The addition of NaCl at high (1 M) concentration competed with the TMPyP-evoked nanomechanical changes. Because the largest amplitude of the changes is induced by the pharmacologically relevant TMPyP concentration range, this porphyrin derivative may be used to tune DNA's structure and properties, hence control the wide array of biomolecular DNA-dependent processes including replication, transcription, condensation and repair., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Hypothesis: Synthetic micro/nanomotors are gaining extensive attention for various biomedical applications (especially in drug delivery) due to their ability to mimic the motion of biological micro/nanoscale swimmers. The feasibility of these applications relies on tight control of propulsion speed, direction, and type of motion (translation, circular, etc.) along with the exerted self-propulsive force. We propose to exploit the variation of both self-propulsion speed and force of active colloids with different patch coverages (with and without supporting layer) for engineering diffusiophoretic micro/nanomotors., Experiments: The microswimmers were designed at various patch coverages (10°, 30°, and 90°) with (Ti/Pt) and without (Pt) an adhesion layer for the catalytic patch through glancing angle metal deposition (GLAD) technique. Mean-square displacement (MSD) analysis was performed to obtain the self-propulsion parameters like speed and angular speed. Using optical tweezers (OT), the self-propulsive force was measured from the force power spectral density., Findings: The findings of our experiments suggest the non-requirement of any adhesion layer preceding the catalyst deposition since the Pt 10° colloidal batch had the maximal self-propulsion speed (4.61±0.3μm/s) and force (345±57fN) for 5% w/v H 2 O 2 fuel concentration. Moreover, the self-propulsion speed and force decreased with increasing patch size, contrary to theoretical estimates. Also, the self-propulsive force obtained from MSD is 2 to 4 times lower in magnitude than the OT based force values. We believe that the self-propelling motion of the micromotors is possibly hindered due to interactions with the surface of the quartz cuvette during the optical microscopic analysis. Further, the MSD is limited to the self-propulsive motion in two dimensions. On the other hand, OT based force measurement involve trapping the particles in the bulk of the solution entirely avoiding the particle-substrate interactions. Hence, OT based force measurements are better than the propulsion velocity based stokes drag force estimates. We believe that this study can lay the foundation in designing efficient micro/nanomotors for translational biomedical applications., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Swathi Sudhakar reports financial support was provided by India Ministry of Science & Technology Department of Science and Technology. Swathi Sudhakar reports financial support was provided by Ministry of Education. Ethayaraja Mani reports financial support was provided by India Ministry of Science & Technology Department of Science and Technology. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper]., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Acoustic tweezers have gained substantial interest in biology, engineering, and materials science for their label-free, precise, contactless, and programmable manipulation of small objects. However, acoustic tweezers cannot independently manipulate multiple microparticles simultaneously. This study introduces acousto-dielectric tweezers capable of independently manipulating multiple microparticles and precise control over intercellular distances and cyclical cell pairing and separation for detailed cell-cell interaction analysis. Our acousto-dielectric tweezers leverage the competition between acoustic radiation forces, generated by standing surface acoustic waves (SAWs), and dielectrophoretic (DEP) forces, induced by gradient electric fields. Modulating these fields allows for the precise positioning of individual microparticles at points where acoustic radiation and DEP forces are in equilibrium. This mechanism enables the simultaneous movement of multiple microparticles along specified paths as well as cyclical cell pairing and separation. We anticipate our acousto-dielectric tweezers to have enormous potential in colloidal assembly, cell-cell interaction studies, disease diagnostics, and tissue engineering.