Direct Intracellular Delivery of Cell Impermeable Probes of Protein Glycosylation Using Nanostraws

Bioorthogonal chemistry is an effective tool for elucidating metabolic pathways and measuring cellular activity, yet its use is currently limited by the difficulty of getting probes past the cell membrane and into the cytoplasm, especially if more complex probes are desired. Here we present a simple and minimally perturbative technique to deliver functional probes of glycosylation into cells by using a nanostructured “nanostraw” delivery system. Nanostraws provide direct intracellular access to cells through fluid conduits that remain small enough to minimize cell perturbation. First, we demonstrate that our platform can deliver an unmodified azidosugar, N-azidoacetylmannosamine, into cells with similar effectiveness to a chemical modification strategy (peracetylation). We then show that the nanostraw platform enables direct delivery of an azidosugar modified with a charged uridine diphosphate group (UDP) that prevents intracellular penetration, thereby bypassing multiple enzymatic processing steps. By effectively removing the requirement for cell permeability from the probe, the nanostraws expand the toolbox of bioorthogonal probes that can be used to study biological processes on a single, easy-to-use platform

Temporally resolved direct delivery of second messengers into cells using nanostraws

Second messengers are biomolecules with the critical role of conveying information to intracellular targets. They are typically membrane-impermeable and only enter cells through tightly regulated transporters. Current methods for manipulating second messengers in cells require preparation of modified cell lines or significant disruptions in cell function, especially at the cell membrane. Here we demonstrate that 100 nm diameter ‘nanostraws’ penetrate the cell membrane to directly modulate second messenger concentrations within cells. Nanostraws are hollow vertical nanowires that provide a fluidic conduit into cells to allow time-resolved delivery of the signaling ion Ca^(2+) without chemical permeabilization or genetic modification, minimizing cell perturbation. By integrating the nanostraw platform into a microfluidic device, we demonstrate coordinated delivery of Ca^(2+) ions into hundreds of cells at the time scale of several seconds with the ability to deliver complex signal patterns, such as oscillations over time. The diffusive nature of nanostraw delivery gives the platform unique versatility, opening the possibility for time-resolved delivery of any freely diffusing molecules

A Model for Emission Yield from Planar Photocathodes Based on Photon-Enhanced Thermionic Emission or Negative-Electron-Affinity Photoemission

A general model is presented for electron emission yield from planar photocathodes that accounts for arbitrary cathode thickness and finite recombination velocities at both front and back surfaces. This treatment is applicable to negative electron affinity emitters as well as positive electron affinity cathodes, which have been predicted to be useful for energy conversion. The emission model is based on a simple one-dimensional steady-state diffusion treatment. The resulting relation for electron yield is used to model emission from thin-film cathodes with material parameters similar to GaAs. Cathode thickness and recombination at the emissive surface are found to strongly affect emission yield from cathodes, yet the magnitude of the effect greatly depends upon the emission mechanism. A predictable optimal film thickness is found from a balance between optical absorption, surface recombination, and emission rate

Hybrid Group IV Nanophotonic Structures Incorporating Diamond Silicon-Vacancy Color Centers

We demonstrate a new approach for engineering group IV semiconductor-based quantum photonic structures containing negatively charged silicon-vacancy (SiV$^-$) color centers in diamond as quantum emitters. Hybrid SiC/diamond structures are realized by combining the growth of nanoand micro-diamonds on silicon carbide (3C or 4H polytype) substrates, with the subsequent use of these diamond crystals as a hard mask for pattern transfer. SiV$^-$ color centers are incorporated in diamond during its synthesis from molecular diamond seeds (diamondoids), with no need for ionimplantation or annealing. We show that the same growth technique can be used to grow a diamond layer controllably doped with SiV$^-$ on top of a high purity bulk diamond, in which we subsequently fabricate nanopillar arrays containing high quality SiV$^-$ centers. Scanning confocal photoluminescence measurements reveal optically active SiV$^-$ lines both at room temperature and low temperature (5 K) from all fabricated structures, and, in particular, very narrow linewidths and small inhomogeneous broadening of SiV$^-$ lines from all-diamond nano-pillar arrays, which is a critical requirement for quantum computation. At low temperatures (5 K) we observe in these structures the signature typical of SiV$^-$ centers in bulk diamond, consistent with a double lambda. These results indicate that high quality color centers can be incorporated into nanophotonic structures synthetically with properties equivalent to those in bulk diamond, thereby opening opportunities for applications in classical and quantum information processing

A Quantum Photonic Interface for Tin-Vacancy Centers in Diamond

The realization of quantum networks critically depends on establishing efficient, coherent light-matter interfaces. Optically active spins in diamond have emerged as promising quantum nodes based on their spin-selective optical transitions, long-lived spin ground states, and potential for integration with nanophotonics. Tin-vacancy (SnV$^{\,\textrm{-}}$) centers in diamond are of particular interest because they exhibit narrow-linewidth emission in nanostructures and possess long spin coherence times at temperatures above 1 K. However, a nanophotonic interface for SnV$^{\,\textrm{-}}$ centers has not yet been realized. Here, we report cavity enhancement of the emission of SnV$^{\,\textrm{-}}$ centers in diamond. We integrate SnV$^{\,\textrm{-}}$ centers into one-dimensional photonic crystal resonators and observe a 40-fold increase in emission intensity. The Purcell factor of the coupled system is 25, resulting in channeling of the majority of photons ($90\%$) into the cavity mode. Our results pave the way for the creation of efficient, scalable spin-photon interfaces based on SnV$^{\,\textrm{-}}$ centers in diamond

Temperature-dependent optical properties of titanium nitride

The refractory metal titanium nitride is promising for high-temperature nanophotonic and plasmonic applications, but its optical properties have not been studied at temperatures exceeding 400 °C. Here, we perform in-situ high-temperature ellipsometry to quantify the permittivity of TiN films from room temperature to 1258 °C. We find that the material becomes more absorptive at higher temperatures but maintains its metallic character throughout visible and near infrared frequencies. X-ray diffraction, atomic force microscopy, and mass spectrometry confirm that TiN retains its bulk crystal quality and that thermal cycling increases the surface roughness, reduces the lattice constant, and reduces the carbon and oxygen contaminant concentrations. The changes in the optical properties of the material are highly reproducible upon repeated heating and cooling, and the room-temperature properties are fully recoverable after cooling. Using the measured high-temperature permittivity, we compute the emissivity, surface plasmon polariton propagation length, and two localized surface plasmon resonance figures of merit as functions of temperature. Our results indicate that titanium nitride is a viable plasmonic material throughout the full temperature range explored

Direct Intracellular Delivery of Cell Impermeable Probes of Protein Glycosylation Using Nanostraws

Bioorthogonal chemistry is an effective tool for elucidating metabolic pathways and measuring cellular activity, yet its use is currently limited by the difficulty of getting probes past the cell membrane and into the cytoplasm, especially if more complex probes are desired. Here we present a simple and minimally perturbative technique to deliver functional probes of glycosylation into cells by using a nanostructured “nanostraw” delivery system. Nanostraws provide direct intracellular access to cells through fluid conduits that remain small enough to minimize cell perturbation. First, we demonstrate that our platform can deliver an unmodified azidosugar, N-azidoacetylmannosamine, into cells with similar effectiveness to a chemical modification strategy (peracetylation). We then show that the nanostraw platform enables direct delivery of an azidosugar modified with a charged uridine diphosphate group (UDP) that prevents intracellular penetration, thereby bypassing multiple enzymatic processing steps. By effectively removing the requirement for cell permeability from the probe, the nanostraws expand the toolbox of bioorthogonal probes that can be used to study biological processes on a single, easy-to-use platform