Single-Molecule Investigation of Load-Dependent Actomyosin Dissociation Kinetics for Cardiac and Slow Skeletal Myosin.

Myosins are ATP-powered, force-generating motor proteins involved in cardiac and muscle contraction. The external load experienced by the myosins modulates and coordinates their function in vivo. Here, this study investigates the tension-sensing mechanisms of rabbit native ß-cardiac myosin (ßM-II) and slow skeletal myosins (SolM-II) that perform in different physiological settings. Using mobile optical tweezers with a square wave-scanning mode, a range of external assisting and resisting loads from 0 to 15 pN is exerted on single myosin molecules as they interact with the actin filament. Influenced of load on specific strongly-bound states in the cross-bridge cycle is examined by adjusting the [ATP]. The results implies that the detachment kinetics of actomyosin ADP.Pi strongly-bound force-generating state are load sensitive. Low assisting load accelerates, while the resisting load hinders the actomyosin detachment, presumably, by slowing both the Pi and ADP release. However, under both high assisting and resisting load, the rate of actomyosin dissociation decelerates. The transition from actomyosin ADP.Pi to ADP state appears to occur with a higher probability for ßM-II than SolM-II. This study interpret that dissociation of at least three strongly-bound actomyosin states are load-sensitive and may contribute to functional diversity among different myosins.

Raman Spectroscopy of Optically Trapped Living Human T Cell Subsets and Monocytes.

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.

Spatiotemporal formation of a single liquid-like condensate and amyloid fibrils of α-synuclein by optical trapping at solution surface.

Liquid-like protein condensates have recently attracted much attention due to their critical roles in biological phenomena. They typically show high fluidity and reversibility for exhibiting biological functions, while occasionally serving as sites for the formation of amyloid fibrils. To comprehend the properties of protein condensates that underlie biological function and pathogenesis, it is crucial to study them at the single-condensate level; however, this is currently challenging due to a lack of applicable methods. Here, we demonstrate that optical trapping is capable of inducing the formation of a single liquid-like condensate of α-synuclein in a spatiotemporally controlled manner. The irradiation of tightly focused near-infrared laser at an air/solution interface formed a condensate under conditions coexisting with polyethylene glycol. The fluorescent dye-labeled imaging showed that the optically induced condensate has a gradient of protein concentration from the center to the edge, suggesting that it is fabricated through optical pumping-up of the α-synuclein clusters and the expansion along the interface. Furthermore, Raman spectroscopy and thioflavin T fluorescence analysis revealed that continuous laser irradiation induces structural transition of protein molecules inside the condensate to ß-sheet rich structure, ultimately leading to the condensate deformation and furthermore, the formation of amyloid fibrils. These observations indicate that optical trapping is a powerful technique for examining the microscopic mechanisms of condensate appearance and growth, and furthermore, subsequent aging leading to amyloid fibril formation.

Deterministic Assembly of Single-Emitter Plasmonic Antenna for Ultrahigh Photoluminescence Enhancement.

Single-emitter nanoantennas play a crucial role in the fabrication of nanosensors and integrated sources. Since the coupling of single emitter to nanoantennas is largely based on stochastic methods, low qualified rate still hinders a massive deployment. Here, we proposed a deterministic, optical-force-driven method to achieve gap-plasmonic photoluminescence enhancement. Two deterministic steps are carried out in sequence: a composite nanoemitter is first synthesized by linking quantum dots to a silica-rapped gold nanoparticle, followed by an optical delivery of the nanoparticle into a nanoaperture in a gold film. We reason that the nanoparticle-in-nanoaperture (NPiNA) structure efficiently couples out-of-plane excitation light into a gap-plasmon via a transverse electromagnetic mode (TEM)-like transmission mode. An in situ photoluminescence measurement demonstrates a 3× brightness as compared to the nanoparticle-on-mirror (NPoM). This approach paves the way toward deterministic positioning of individual nanoparticles for a wide range of applications on nanophotonics structures on-a-chip.

Enhanced Plasmonic Trapping and Fluorescent Emission of Nitrogen-Vacancy Nanodiamonds Using a High-Efficiency Nanofocusing Device.

Fluorescent nanodiamonds (FNDs) with nitrogen-vacancy centers are pivotal for advancing quantum photonics and imaging through deterministic quantum state manipulation. However, deterministic integration of quantum emitters into photonic devices remains a challenge due to the need for high coupling efficiency and Purcell enhancement. We report a deterministic FND-integrated nanofocusing device achieved by assembling FNDs at a plasmonic waveguide tip through plasmonic-enhanced optical trapping. This technique not only increases the emission rate by 58.6 times compared to isolated FNDs but also preferentially directs radiation into the waveguide at a rate 5.3 times higher than that into free space, achieving an exceptional figure-of-merit of ∼3000 for efficient energy transfer. Our findings represent a significant step toward deterministic integration in quantum imaging and communication, opening new avenues for quantum technology advancements.

High Stability and Low Power Nanometric Bio-Objects Trapping through Dielectric-Plasmonic Hybrid Nanobowtie.

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/µm2 while preserving the temperature, chemical-biological properties, and shape of the biological sample.

Optical trapping-induced crystallization promoted by gold and silicon nanoparticles.

This study investigates the promotion of sodium chlorate (NaClO3) crystallization through optical trapping, enhanced by the addition of gold nanoparticles (AuNPs) and silicon nanoparticles (SiNPs). Using a focused laser beam at the air-solution interface of a saturated NaClO3 solution with AuNPs or SiNPs, the aggregates of these particles were formed at the laser focus, the nucleation and growth of metastable NaClO3 (m-NaClO3) crystals were induced. Continued laser irradiation caused these m-NaClO3 crystals to undergo repeated cycles of growth and dissolution, eventually transitioning to a stable crystal form. Our comparative analysis showed that AuNPs, due to their significant heating due to higher photon absorption efficiency, caused more pronounced size fluctuations in m-NaClO3 crystals compared to the stable behavior observed with SiNPs. Interestingly, the maximum diameter of the m-NaClO3 crystals that appeared during the size fluctuation step was consistent, regardless of nanoparticle type, concentration, or size. The crystallization process was also promoted by using polystyrene nanoparticles, which have minimal heating and electric field enhancement, suggesting that the reduction in activation energy for nucleation at the particle surface is a key factor. These findings provide critical insights into the mechanisms of laser-induced crystallization, emphasizing the roles of plasmonic heating, particle surfaces, and optical forces.

Unveiling the Influence of Brightness Heterogeneity in Fluorescence Correlation Spectroscopy of Perovskite Nanocrystals.

Nanoparticles (NPs), including perovskite nanocrystals (PNCs) with single photon purity, present challenges in fluorescence correlation spectroscopy (FCS) studies due to their distinct photoluminescence (PL) behaviors. In particular, the zero-time correlation amplitude [g2(0)] and the associated diffusion timescale (τD) of their FCS curves show substantial dependency on pump intensity (IP). Optical saturation inadequately explains the origin of this FCS phenomenon in NPs, thus setting them apart from conventional dye molecules, which do not manifest such behavior. This observation is apparently attributed to either photo-brightening or optical trapping, both lead to increased NP occupancy (N) in the excitation volume, consequently reducing the g2(0) amplitude [since g2(0) α 1/N] at high IP. However, an advanced FCS study utilizing alternating laser excitation at two different intensities dismisses such possibilities. Further investigation into single-particle blinking behaviors as a function of IP reveals that the intensity dependence of g2(0) primarily arises from the brightness heterogeneity prevalent in almost all types of NPs. This report delves into the complexities of the photophysical properties of NPs and their adverse impacts on FCS studies.

Modular Assembly of Metamaterials Using Light Gradients.

This is a report on a pilot study that tests the feasibility of assembling photonic metamaterials (PMs) using light gradient forces. Following a strategy that works like modular construction, light gradient forces, produced by a tightly focused, 1D standing wave optical trap, time-multiplexed across a 2D lattice are used to assemble voxels consisting of prefabricated, monodispersed nanoparticles (NPs) with radii ranging from 30 to 500 nm into 3D structures on a hydrogel scaffold. Hundreds of NPs can be manipulated concurrently into a complex heterogeneous voxel this way, and then the process can be repeated by stitching together voxels to form a metamaterial of any size, shape, and constituency although imperfectly. Imperfections introduce random phase shifts and amplitude variations that can have an adverse effect on the band structure. Regardless, PMs are created this way using two different dielectric NPs, polystyrene and rutile, and then the near-infrared performance for each is analyzed with angle-, wavelength-, and polarization-dependent reflection spectroscopy. The cross-polarized spectra show evidence of a resonance peak. Interestingly, whereas the line shape from the polystyrene array is symmetric, the rutile array is not, which may be indicative of Fano resonance. So, even with the structural defects, reflection spectroscopy reveals a resonance.

Optical Tweezing Terahertz Probing for a Single Metal Nanoparticle.

Recently, extensive research has been reported on the detection of metal nanoparticles using terahertz waves, due to their potential for efficient and nondestructive detection of chemical and biological samples without labeling. Resonant terahertz nanoantennas can be used to detect a small amount of molecules whose vibrational modes are in the terahertz frequency range with high sensitivity. However, the positioning of target molecules is critical to obtaining a reasonable signal because the field distribution is inhomogeneous over the antenna structure. Here, we combine an optical tweezing technique and terahertz spectroscopy based on nanoplasmonics, resulting in extensive controllable tweezing and sensitive detection at the same time. We observed optical tweezing of a gold nanoparticle and detected it with terahertz waves by using a single bowtie nanoantenna. Furthermore, the calculations confirm that molecular fingerprinting is possible by using our technique. This study will be a prestep of biomolecular detection using gold nanoparticles in terahertz spectroscopy.

Controlled Second Harmonic Generation with Optically Trapped Lithium Niobate Nanoparticles.

We report the second harmonic generation (SHG) response from a single 34 nm diameter lithium niobate nanoparticle. The experimental setup involves a first beam devoted to the optical trapping of single nanoparticles, whereas a second arm involves a femtosecond laser source leading to the SHG emission from the trapped nanoparticles. SHG operation where one to three nanoparticles are present in the optical trap is first demonstrated, highlighting the transition between coherent and incoherent SHG, the latter known as hyper-Rayleigh scattering (HRS). With a spatial light modulator moving the optical trap in and out of the focus of the femtosecond beam, the SHG intensity is switched back and forth between a low and a high level. This controlled operation opens new avenues for nanoparticle characterization and applications in sensing or communication and information technologies and constitutes the first step in the design of active substrateless metasurfaces.

Effect of the Photoexcitation Wavelength and Polarization on the Generated Heat by a Nd-Doped Microspinner at the Microscale.

Thermal control at small scales is critical for studying temperature-dependent biological systems and microfluidic processes. Concerning this, optical trapping provides a contactless method to remotely study microsized heating sources. This work introduces a birefringent luminescent microparticle of NaLuF4:Nd3+ as a local heater in a liquid system. When optically trapped with a circularly polarized laser beam, the microparticle rotates and heating is induced through multiphonon relaxation of the Nd3+ ions. The temperature increment in the surrounding medium is investigated, reaching a maximum heating of ≈5 °C within a 30 µm radius around the static particle under 51 mW laser excitation at 790 nm. Surprisingly, this study reveals that the particle's rotation minimally affects the temperature distribution, contrary to the intuitive expectation of liquid stirring. The influence of the microparticle rotation on the reduction of heating transfer is analyzed. Numerical simulations confirm that the thermal distribution remains consistent regardless of spinning. Instead, the orientation-dependence of the luminescence process emerges as a key factor responsible for the reduction in heating. The anisotropy in particle absorption and the lag between the orientation of the particle and the laser polarization angle contribute to this effect. Therefore, caution must be exercised when employing spinning polarization-dependent luminescent particles for microscale thermal analysis using rotation dynamics.

Nanoscale Helical Optical Force for Determining Crystal Chirality.

The deterministic control of material chirality has been a sought-after goal. As light possesses intrinsic chirality, light-matter interactions offer promising avenues for achieving non-contact, enantioselective optical induction, assembly, or sorting of chiral entities. However, experimental validations are confined to the microscale due to the limited strength of asymmetrical interactions within sub-diffraction limit ranges. In this study, a novel approach is presented to facilitate chirality modulation through chiral crystallization using a helical optical force field originating from localized nanogap surface plasmon resonance. The force field emerges near a gold trimer nanogap and is propelled by linear and angular momentum transfer from the incident light to the resonant nanogap plasmon. By employing Gaussian and Laguerre-Gaussian incident laser beams, notable enantioselectivity is achieved through low-power plasmon-induced chiral crystallization of an organic compound-ethylenediamine sulfate. The findings provide new insights into chirality transmission orchestrated by the exchange of linear and angular momentum between light and nanomaterials.

Efficient and Shape-Sensitive Manipulation of Nanoparticles by Quasi-Bound States in the Continuum Modes in All-Dielectric Metasurfaces.

Current optical tweezering techniques are actively employed in the manipulation of nanoparticles, e.g., biomedical cells. However, there is still huge room for improving the efficiency of manipulating multiple nanoparticles of the same composition but different shapes. In this study, we designed an array of high-index all-dielectric disk antennas, each with an asymmetric open slot for such applications. Compared with the plasmonic counterparts, this all-dielectric metasurface has no dissipation loss and, thus, circumvents the Joule heating problem of plasmonic antennas. Furthermore, the asymmetry-induced excitation of quasi-bound states in continuum (QBIC) mode with a low-power intensity (1 mW/µm2) incidence imposes an optical gradient force of -0.31 pN on 8 nm radius nanospheres, which is four orders of magnitude stronger than that provided by the Fano resonance in plasmonic antenna arrays, and three orders of magnitude stronger than that by the Mie resonance in the same metasurface without any slot, respectively. This asymmetry also leads to the generation of large optical moments. At the QBIC resonance wavelength, a value of 88.3 pN-nm will act on the nanorods to generate a rotational force along the direction within the disk surface but perpendicular to the slot. This will allow only nanospheres but prevent the nanorods from accurately entering into the slots, realizing effective sieving between the nanoparticles of the two shapes.

HoloTile light engine: new digital holographic modalities and applications.

HoloTile is a patented computer generated holography approach with the aim of reducing the speckle noise caused by the overlap of the non-trivial physical extent of the point spread function in Fourier holographic systems from adjacent frequency components. By combining tiling of phase-only of rapidly generated sub-holograms with a PSF-shaping phase profile, each frequency component-or output 'pixel'- in the Fourier domain is shaped to a desired non-overlapping profile. In this paper, we show the high-resolution, speckle-reduced reconstructions that can be achieved with HoloTile, as well as present new HoloTile modalities, including an expanded list of PSF options with new key properties. In addition, we discuss numerous applications for which HoloTile, its rapid hologram generation, and the new PSF options may be an ideal fit, including optical trapping and manipulation of particles, volumetric additive printing, information transfer and quantum communication.

Optical Trapping and Fast Discrimination of Label-Free Bacteriophages at the Single Virion Level.

There is a recent resurgence of interest in phage therapy (the therapeutic use of bacterial viruses) as an approach to eliminating difficult-to-treat infections. However, existing approaches for therapeutic phage selection and virulence testing are time-consuming, host-dependent, and facing reproducibility issues. Here, this study presents an innovative approach wherein integrated resonant photonic crystal (PhC) cavities in silicon are used as optical nanotweezers for probing and manipulating single bacteria and single virions with low optical power. This study demonstrates that these nanocavities differentiate between a bacterium and a phage without labeling or specific surface bioreceptors. Furthermore, by tailoring the spatial extent of the resonant optical mode in the low-index medium, phage distinction across phenotypically distinct phage families is demonstrated. The work paves the road to the implementation of optical nanotweezers in phage therapy protocols.

Enantioselective Optical Trapping of Multiple Pairs of Enantiomers by Focused Hybrid Polarized Beams.

Enantiomers (opposite chiral molecules) usually exhibit different effects when interacting with chiral agents, thus the identification and separation of enantiomers are of importance in pharmaceuticals and agrochemicals. Here an optical approach is proposed to enantioselective trapping of multiple pairs of enantiomers by a focused hybrid polarized beam. Numerical results indicate that such a focused beam shows multiple local optical chirality of opposite signs in the focal plane, and can trap the corresponding enantiomers near the extreme value of optical chirality density according to the handedness of enantiomers. The number and positions of trapped enantiomers can be changed by altering the value and sign of polarization orders of hybrid polarized beams, respectively. The key to realizing enantioselective optical trapping of enantiomers is that the chiral optical force exerted on enantiomers in this focused field is stronger than the achiral optical force. The results provide insight into the optical identification and separation of multiple pairs of enantiomers and will find applications in chiral detection and sensing.

Light-sheet Raman tweezers for whole-cell biochemical analysis of functional red blood cells.

Micro-Raman spectroscopy has emerged as one of the foremost techniques for analyzing biological cells in recent years due to its non-destructive nature and high spatial resolution. The development of optical tweezers has eased the research on biological cells as they confine living cells and organisms in the optical trap without causing much damage. Combining optical tweezers with Raman spectroscopy has opened a wide range of applications in the biomedical field as it facilitates biochemical analysis of biological samples by maintaining in-vivo conditions. Herein, we developed a light sheet-based optical tweezer that traps red blood cells (RBCs) at a very low power density spread across the whole cell, otherwise impossible with conventional optical tweezers. Furthermore, it is combined with micro-Raman spectroscopy to perform whole-cell biochemical analysis for the first time. Raman spectra of individual RBCs recorded under the line focal spot excitation are of superior quality and lack spectral signatures of photo-oxidation and heme aggregation, which is common in point focal spot excitations.

Droplet-based optical trapping for cell separation in mock forensic samples.

Optical tweezers have a wide range of uses for mechanical manipulation of objects in the microscopic range. This includes both living and static cells in a variety of biomedical and research applications. Single-focus optical tweezers, formed by focusing a laser beam through a high numerical aperture immersion objective, create a significant force, which enables controlled transport of a variety of different cell types and morphologies in three dimensions. Optical tweezers have been previously reported to capture and separate spermatozoa from a reconstituted simulated postcoital sample. We report herein the development of a simplified, more efficient cell transfer protocol that can separate and isolate both spermatozoa as well as leukocytes, with similar efficiencies as those previously reported. The new cell transfer method was used to separate sperm cells from a reconstituted mixture of spermatozoa and vaginal epithelial cells, with complete STR profiles developed from 50 cells with little evidence of contribution from the female contributor to the mixture. This modified protocol was then used to separate 21 samples of enriched leukocytes, with trapped cells ranging from 5 to 22 cells. Complete STR profiles were developed from as few as 10 leukocytes. Thus, with minimal sample preparation and a short trapping time, this method has the potential to provide an alternative to traditional differential extraction methods for separation of sperm:nonsperm mixtures while also providing versatility for separation of cells with differing morphologies.

Single Elementary Charge Fluctuations on Nanoparticles in Aqueous Solution.

100 years ago, in 1923, the Nobel prize in physics was awarded for measurement of the unit charge. In addition to a profound impact on contemporary physics, this discovery has reshaped our understanding of charge-based interactions in chemistry and biology, ranging from oxidation and ionization to protein folding and metabolism. In a liquid, the discrete nature of the electric charge becomes prominent at the nanoscale when a charge carrier is exchanged between a molecule or a nanoparticle and the surrounding medium. However, our ability to observe the dynamics of such interactions at the level of a single elementary charge is limited due to the abundance of ions in water. Here, we report on the observation of single binding-unbinding events with elementary charge resolution at the surface of a nanoparticle suspended in water. Discrete steps in the electrical charge are revealed by analyzing the motion of optically trapped nanoparticles under the influence of an applied sinusoidal electric field. The measurements are sufficiently fast and long to observe individual (dis)charging events that occur on average every 3 s. Our results offer prospective routes for studying the dynamics of diverse chemical and biological phenomena on the nanoscale with elementary charge resolution.