Ligand functionalization of titanium nanopattern enables the analysis of cell-ligand interactions by super-resolution microscopy
The spatiotemporal aspects of early signaling events during interactions between cells and their environment dictate multiple downstream outcomes. While advances in nanopatterning techniques have allowed the isolation of these signaling events, a major limitation of conventional nanopatterning methods is its dependence on gold (Au) or related materials that plasmonically quench fluorescence and, thus, are incompatible with super-resolution fluorescence microscopy. Here we describe a novel method that integrates nanopatterning with single-molecule resolution fluorescence imaging, thus enabling mechanistic dissection of molecular-scale signaling events in conjunction with nanoscale geometry manipulation. Our method exploits nanofabricated titanium (Ti) whose oxide (TiO2) is a dielectric material with no plasmonic effects. We describe the surface chemistry for decorating specific ligands such as cyclo-RGD (arginine, glycine and aspartate: a ligand for fibronectin-binding integrins) on TiO2 nanoline and nanodot substrates, and demonstrate the ability to perform dual-color super-resolution imaging on these patterns. Ti nanofabrication is similar to other metallic materials like Au, while the functionalization of TiO2 is relatively fast, safe, economical, easy to set up with commonly available reagents, and robust against environmental parameters such as humidity. Fabrication of nanopatterns takes ~2-3 d, preparation for functionalization ~1.5-2 d, and functionalization 3 h, after which cell culture and imaging experiments can be performed. We suggest that this method may facilitate the interrogation of nanoscale geometry and force at single-molecule resolution, and should find ready applications in early detection and interpretation of physiochemical signaling events at the cell membrane in the fields of cell biology, immunology, regenerative medicine, and related fields.
On-Demand Fully Enclosed Superhydrophobic-Optofluidic Devices Enabled by Microstereolithography
Superhydrophobic surface-based optofluidics have been introduced to biosensors and unconventional optics with unique advantages, such as low light loss and power consumption. However, most of these platforms were made with planar-like microstructures and nanostructures, which may cause bonding issues and result in significant waveguide loss. Here, we introduce a fully enclosed superhydrophobic-based optofluidics system, enabled by a one-step microstereolithography procedure. Various microstructured cladding designs with a feature size down to 100 Î¼m were studied and a "T-type" overhang design exhibits the lowest optical loss, regardless of the excitation wavelength. Surprisingly, the optical loss of superhydrophobic-based optofluidics is not solely decided by the solid area fraction at the solid/water/air interface, but also the cross-section shape and the effective cladding layer composition. We show that this fully enclosed optofluidic system can be used for CRISPR-labeled quantum dot quantification, intended for in vitro and in vivo CRISPR therapeutics.
Broadband Liquid Crystal Tunable Metasurfaces in the Visible: Liquid Crystal Inhomogeneities across the Metasurface Parameter Space
Optical metasurfaces - planar nanostructured devices that can arbitrarily tailor the wavefront of light - may be reconfigured by changing their dielectric environment. The application of external stimuli to liquid crystals is a particularly promising means of tuning the optical properties of embedded metasurfaces because of liquid crystals' large and broadband optical anisotropy. However, the detailed behavior of liquid crystals immediately adjacent to the nanostructured meta-atoms elements is often overlooked, despite the optics of the device depending sensitively on this behavior (e.g., the spectral position of the meta-atom resonances). This is of increasing concern as the wavelength of operation further approaches the short-wavelength end of the visible spectrum and, therefore, the length scale of the inhomogeneities in the liquid crystal director field. In this manuscript, we undertake a fully comprehensive study, across the metasurface geometrical parameter space, of broadband (450-700 nm) all-dielectric liquid crystal tunable metasurfaces operating in the visible. Through combined experimental characterization, liquid crystal modeling, and optical simulations, we reveal and quantify the improved accuracy with which the optical properties of the liquid crystal tunable metasurfaces may be described, and identify the underlying physical mechanism: the three-dimensional spatial overlap of the liquid crystal director field and metasurface optical near fields in the vicinity of the meta-atoms.
Contribution of Ferromagnetic Medium to the Output of Triboelectric Nanogenerators Derived from Maxwell's Equations
Triboelectric nanogenerators (TENGs) use the displacement current as a driving force to convert mechanical energy into electric power, which has made great contributions to micro-nano energy harvesting, self-powered systems, and the sustainable development of mankind. To date, it is accepted that the output of TENGs is only dependent on the polarization effect. This study reveals that this view is incomplete and, in reality, the magnetization effect also makes a significant contribution to the output of TENGs. For the first time, a novel insight on the output of TENGs is discovered through the theoretical derivation and analysis of Maxwell's equations in ferromagnetic medium. Experimentally, TENGs based on ferromagnetic media are constructed, which exhibit higher output than that of non-ferromagnetic media based. Interestingly, the output behavior of ferromagnetic media based TENGs is strongly related to the external magnetic field ambient, which is well demonstrated. The discovered output characteristics of TENGs are precisely derived from the working principle of TENGs, simultaneously, a completed and unified theoretical system is constructed for TENGs. This significant discovery and theory will be an indispensable supplement to the existing research on TENGs and also provide a general guidance and deeper understanding of the TENG.
Polarization-Insensitive Medium-Switchable Holographic Metasurfaces
The adoption of metasurfaces has led to revolutionary advances in holography due to improved compactness, integrability, and performance. Switchable meta-holograms projecting different replay field images in a controllable manner are highly desirable. Still, existing technologies generally rely on the use of polarized light and additional optics to facilitate switching. Consequently, the potential benefits afforded through the use of metasurfaces are limited both by the system complexity and a fixed relationship between the optical input and output. In this manuscript, we demonstrate polarization-insensitive metasurfaces encoding arbitrary and independent holograms, which can be switched between by changing the refractive index of the infiltration medium while maintaining identical illumination conditions. By sidestepping the requirements for high-performance light sources, switching optics, or delicate alignment, this approach points toward ultracompact and cost-effective switchable meta-holograms for various practical applications, such as holographic image projection, eye-perceptible sensors, optical information storage, processing, and security.
Kirigami Engineering-Nanoscale Structures Exhibiting a Range of Controllable 3D Configurations
Kirigami structures provide a promising approach to transform flat films into 3D complex structures that are difficult to achieve by conventional fabrication approaches. By designing the cutting geometry, it is shown that distinct buckling-induced out-of-plane configurations can be obtained, separated by a sharp transition characterized by a critical geometric dimension of the structures. In situ electron microscopy experiments reveal the effect of the ratio between the in-plane cut size and film thickness on out-of-plane configurations. Moreover, geometrically nonlinear finite element analyses (FEA) accurately predict the out-of-plane modes measured experimentally, their transition as a function of cut geometry, and provide the stress-strain response of the kirigami structures. The combined computational-experimental approach and results reported here represent a step forward in the characterization of thin films experiencing buckling-induced out-of-plane shape transformations and provide a path to control 3D configurations of micro- and nanoscale buckling-induced kirigami structures. The out-of-plane configurations promise great utility in the creation of micro- and nanoscale systems that can harness such structural behavior, such as optical scanning micromirrors, novel actuators, and nanorobotics. This work is of particular significance as the kirigami dimensions approach the sub-micrometer scale which is challenging to achieve with conventional micro-electromechanical system technologies.
Inverse design of metasurfaces with non-local interactions
Conventional metasurfaces have demonstrated efficient wavefront manipulation by using thick and high-aspect-ratio nanostructures in order to eliminate interactions between adjacent phase-shifter elements. Thinner-than-wavelength dielectric metasurfaces are highly desirable because they can facilitate fabrication and integration with both electronics and mechanically tunable platforms. Unfortunately, because their constitutive phase-shifter elements exhibit strong electromagnetic coupling between neighbors, the design requires a global optimization methodology that considers the non-local interactions. Here, we propose a global evolutionary optimization approach to inverse design non-local metasurfaces. The optimal designs are experimentally validated, demonstrating the highest efficiencies for the thinnest transmissive metalenses reported to-date for visible light. In a departure from conventional design methods based on the search of a library of pre-determined and independent meta-atoms, we take full advantage of the strong interactions among nanoresonators to improve the focusing efficiency of metalenses and demonstrate that efficiency improvements can be obtained by lowering the metasurface filling factors.
The CAR T-Cell Mechanoimmunology at a Glance
Chimeric antigen receptor (CAR) T-cell transfer is a novel paradigm of adoptive T-cell immunotherapy. When coming into contact with a target cancer cell, CAR T-cell forms a nonclassical immunological synapse with the cancer cell and dynamically orchestrates multiple critical forces to commit cytotoxic immune function. Such an immunologic process involves a force transmission in the CAR and a spatiotemporal remodeling of cell cytoskeleton to facilitate CAR activation and CAR T-cell cytotoxic function. Yet, the detailed understanding of such mechanotransduction at the interface between the CAR T-cell and the target cell, as well as its molecular structure and signaling, remains less defined and is just beginning to emerge. This article summarizes the basic mechanisms and principles of CAR T-cell mechanoimmunology, and various lessons that can be comparatively learned from interrogation of mechanotransduction at the immunological synapse in normal cytotoxic T-cell. The recent development and future application of novel bioengineering tools for studying CAR T-cell mechanoimmunology is also discussed. It is believed that this progress report will shed light on the CAR T-cell mechanoimmunology and encourage future researches in revealing the less explored yet important mechanosensing and mechanotransductive mechanisms involved in CAR T-cell immuno-oncology.
Integrin nanoclusters can bridge thin matrix fibres to form cell-matrix adhesions
Integrin-mediated cell-matrix adhesions are key to sensing the geometry and rigidity of extracellular environments and influence vital cellular processes. In vivo, the extracellular matrix is composed of fibrous arrays. To understand the fibre geometries that are required for adhesion formation, we patterned nanolines of various line widths and arrangements in single, crossing or paired arrays with the integrin-binding peptide Arg-Gly-Asp. Single thin lines (width â‰¤30â€‰nm) did not support cell spreading or formation of focal adhesions, despite the presence of a high density of Arg-Gly-Asp, but wide lines (>40â€‰nm) did. Using super-resolution microscopy, we observed stable, dense integrin clusters formed on parallel (within 110â€‰nm) or crossing thin lines (mimicking a matrix mesh) similar to those on continuous substrates. These dense clusters bridged the line pairs by recruiting activated but unliganded integrins, as verified by integrin mutants unable to bind ligands that coclustered with ligand-bound integrins when present in an active extended conformation. Thus, in a fibrous extracellular matrix mesh, stable integrin nanoclusters bridge between thin (â‰¤30â€‰nm) matrix fibres and bring about downstream consequences of cell motility and growth.
Ultrathin transmissive metasurfaces for multi-wavelength optics in the visible