Isoelectric Focusing in a Silica Nanofluidic Channel: Effects of Electromigration and Electroosmosis

Isoelectric focusing of proteins in a silica nanofluidic channel filled with citric acid and disodium phosphate buffers is investigated via numerical simulation. Ions in the channel migrate in response to (i) the electric field acting on their charge and (ii) the bulk electroosmotic flow (which is directed toward the cathode). Proteins are focused near the low pH (anode) end when the electromigration effect is more significant and closer to the high pH (cathode) end when the electroosmotic effect dominates. We simulate the focusing behavior of Dylight labeled streptavidin (Dyl-Strep) proteins in the channel, using a relationship between the protein’s charge and pH measured in a previous experiment. Protein focusing results compare well to previous experimental measurements. The effect of some key parameters, such as applied voltage, isoelectric point (pI), bulk pH, and bulk conductivity, on the protein trapping behavior in a nanofluidic channel is examined

Site-Dependent Luminescence and Thermal Stability of Eu²⁺ Doped Fluorophosphate toward White LEDs for Plant Growth

Eu²⁺ activated fluorophosphate Ba₃GdNa­(PO₄)₃F (BGNPF) with blue and red double-color emitting samples were prepared via a solid-state method in a reductive atmosphere. Their crystal structure and cationic sites were identified in light of X-ray diffraction pattern Rietveld refinement. Three different Ba²⁺ sites, coordinated by six O atoms referred to as Ba1, two F and five O atoms as Ba2, and two F and six O atoms as Ba3, were partially substituted by Eu²⁺. Photoluminescence emission (PL) and excitation (PLE) spectra of phosphor BGNPF:Eu²⁺ along with the lifetimes were characterized at the liquid helium temperature (LHT), which further confirm the existence of three Eu²⁺ emitting centers resulting in 436, 480, and 640 nm emission from the 5d → 4f transitions of Eu²⁺ in three different Ba²⁺ crystallographic sites. These emissions overlap with the absorption spectra of carotenoids and chlorophylls from plants, which could directly promote the photosynthesis. Temperature-dependent PL spectra were used to investigate the thermal stability of phosphor, which indicates that the PL intensity of BGNPF:0.9% Eu²⁺ with optimal composition at 150 °C still keeps 60% of its PL intensity at room temperature, in which blue emission has higher thermal-stability than the red emission. Furthermore, the approaching white LED devices have also been manufactured with a 365 nm n-UV LED chip and present phosphor, which make operators more comfortable than that of the plant growth purple emitting LEDs system composed of blue and red light. Results indicate that this phosphor is an attractive dual-responsive candidate phosphor in the application n-UV light-excited white LEDs for plant growth

Stationary Chemical Gradients for Concentration Gradient-Based Separation and Focusing in Nanofluidic Channels

Previous work has demonstrated the simultaneous concentration and separation of proteins via a stable ion concentration gradient established within a nanochannel (Inglis Angew. Chem., Int. Ed. 2001, 50, 7546−7550). To gain a better understanding of how this novel technique works, we here examine experimentally and numerically how the underlying electric potential controlled ion concentration gradients can be formed and controlled. Four nanochannel geometries are considered. Measured fluorescence profiles, a direct indicator of ion concentrations within the Tris–fluorescein buffer solution, closely match depth-averaged fluorescence profiles calculated from the simulations. The simulations include multiple reacting species within the fluid bulk and surface wall charge regulation whereby the deprotonation of silica-bound silanol groups is governed by the local pH. The three-dimensional system is simulated in two dimensions by averaging the governing equations across the (varying) nanochannel width, allowing accurate numerical results to be generated for the computationally challenging high aspect ratio nanochannel geometries. An electrokinetic circuit analysis is incorporated to directly relate the potential drop across the (simulated) nanochannel to that applied across the experimental chip device (which includes serially connected microchannels). The merit of the thick double layer, potential-controlled concentration gradient as a particle focusing and separation tool is discussed, linking this work to the previously presented protein trapping experiments. We explain why stable traps are formed when the flow is in the opposite direction to the concentration gradient, allowing particle separation near the low concentration end of the nanochannel. We predict that tapered, rather than straight nanochannels are better at separating particles of different electrophoretic mobilities

Graphene Oxide Thin Film with Dual Function Integrated into a Nanosandwich Device for in Vivo Monitoring of Interleukin‑6

Graphene oxide (GO), with its exceptional physical and chemical properties and biocompatibility, holds a tremendous potential for sensing applications. In this study, GO, acting both as the electron-transfer bridge and the signal reporter, was attached on the interface to develop a label-free electrochemical nanosandwich device for detection of interleukin-6 (IL-6). First, a single layer of GO was covalently modified on gold electrodes, followed by attachment of anti-IL-6 capture antibody to form the sensing interface. The 4-aminophenyl phosphorylcholine was further attached to the surface of GO to minimize nonspecific protein adsorption. For reporting the presence of analyte, the anti-IL-6 detection antibody was covalently modified to the GO, which has been integrated with the redox probe Nile blue (NB). Finally, a nanosandwich assay was fabricated on gold surfaces for detection of IL-6 on the basis of the electrochemical signal of NB. The prepared nanosandwiches demonstrated high selectivity and stability for detection of IL-6 over the range of 1–300 pg mL^–1 with the lowest detectable concentration of 1 pg mL^–1. The device was successfully used for monitoring of IL-6 secretion in RAW cells and live mice. By tailoring the GO surface with functional components, such devices were able to detect the analyte in vivo without causing inflammatory response

“Turn-on” Fluorescent Aptasensor Based on AIEgen Labeling for the Localization of IFN‑γ in Live Cells

We report an aggregation-induced emission fluorogen (AIEgen)-based turn-on fluorescent aptasensor able to detect the ultrasmall concentration of intracellular IFN-γ. The aptasensor consists of an IFN-γ aptamer labeled with a fluorogen with a typical aggregation-induced emission (AIE) characteristic, which shows strong red emission only in the presence of IFN-γ. The aptasensor is able to effectively monitor intracellular IFN-γ secretion with the lowest detection limit of 2 pg mL^‑1, and it is capable of localizing IFN-γ in live cells during secretion, with excellent cellular permeability and biocompatibility as well as low cytotoxicity. This probe is able to localize the intracellular IFN-γ at a low concentration <10 pg mL^‑1, and it is successfully used for real-time bioimaging. This simple and highly sensitive sensor may enable the exploration of cytokine pathways and their dynamic secretion process in the cellular environment. It provides a universal sensing platform for monitoring a spectrum of molecules secreted by cells

Development of Bright and Biocompatible Nanoruby and Its Application to Background-Free Time-Gated Imaging of G‑Protein-Coupled Receptors

At the forefront of developing fluorescent probes for biological imaging applications are enhancements aimed at increasing their brightness, contrast, and photostability, especially toward demanding applications of single-molecule detection. In comparison with existing probes, nanorubies exhibit unlimited photostability and a long emission lifetime (∼4 ms), which enable continuous imaging at single-particle sensitivity in highly scattering and fluorescent biological specimens. However, their wide application as fluorescence probes has so far been hindered by the absence of facile methods for scaled-up high-volume production and molecularly specific targeting. The present work encompasses the large-scale production of colloidally stable nanoruby particles, the demonstration of their biofunctionality and negligible cytotoxicity, as well as the validation of its use for targeted biomolecular imaging. In addition, optical characteristics of nanorubies are found to be comparable or superior to those of state-of-the-art quantum dots. Protocols of reproducible and robust coupling of functional proteins to the nanoruby surface are also presented. As an example, NeutrAvidin-coupled nanoruby show excellent affinity and specificity to μ-opioid receptors in fixed and live cells, allowing wide-field imaging of G-protein coupled receptors with single-particle sensitivity