Site-selective heat boosting electrochemiluminescence for single cell imaging.

In operando visualization of local electrochemical reactions provides mechanical insights into the dynamic transport of interfacial charge and reactant/product. Electrochemiluminescence is a crossover technique that quantitatively determines Faraday current and mass transport in a straightforward manner. However, the sensitivity is hindered by the low collision efficiency of radicals and side reactions at high voltage. Here, we report a site-selective heat boosting electrochemiluminescence microscopy. By generating a micron-scale heat point in situ at the electrode-solution interface, we achieved an enhancement of luminescence intensity up to 63 times, along with an advance of 0.2 V in applied voltage. Experimental results and finite element simulation demonstrate that the fundamental reasons are accelerated reaction rate and thermal convection via a photothermal effect. The concentrated electrochemiluminescence not only boosts the contrast of single cells by 20.54 times but also enables the site-selective cell-by-cell analysis of the heterogeneous membrane protein abundance. This electrochemical visualization method has great potential in the highly sensitive and selective analysis of local electron transfer events.

An In Situ Investigation of the Protein Corona Formation Kinetics of Single Nanomedicine Carriers by Self-Regulated Electrochemiluminescence Microscopy.

Protein coronas are present extensively at the bio-nano interface due to the natural adsorption of proteins onto nanomaterials in biological fluids. Aside from the robust property of nanoparticles, the dynamics of the protein corona shell largely define their chemical identity by altering interface properties. However, the soft coronas are normally complex and rapidly changing. To real-time monitor the entire formation, we report here a self-regulated electrochemiluminescence (ECL) microscopy based on the interaction of the Ru(bpy)3 3+ with the nanoparticle surface. Thus, the heterogeneity of the protein corona is in situ observed in single nanoparticle "cores" before and after loading drugs in nanomedicine carriers. The label-free, optical stable and dynamic ECL microscopy minimize misinterpretations caused by the variation of nanoparticle size and polydispersity. Accordingly, the synergetic actions of proteins and nanoparticles properties are uncovered by chemically engineered protein corona. After comparing the protein corona formation kinetics in different complex systems and different nanomedicine carriers, the universality and accuracy of this technique were well demonstrated via the protein corona formation kinetics curves regulated by competitive adsorption of Ru(bpy)3 3+ and multiple proteins on surface of various carriers. The work is of great significance for studying bio-nano interface in drug delivery and targeted cancer treatment.

A Review of Smart Superwetting Surfaces Based on Shape-Memory Micro/Nanostructures.

Bioinspired smart superwetting surfaces with special wettability have aroused great attention from fundamental research to technological applications including self-cleaning, oil-water separation, anti-icing/corrosion/fogging, drag reduction, cell engineering, liquid manipulation, and so on. However, most of the reported smart superwetting surfaces switch their wettability by reversibly changing surface chemistry rather than surface microstructure. Compared with surface chemistry, the regulation of surface microstructure is more difficult and can bring novel functions to the surfaces. As a kind of stimulus-responsive material, shape-memory polymer (SMP) has become an excellent candidate for preparing smart superwetting surfaces owing to its unique shape transformation property. This review systematically summarizes the recent progress of smart superwetting SMP surfaces including fabrication methods, smart superwetting phenomena, and related application fields. The smart superwettabilities, such as superhydrophobicity/superomniphobicity with tunable adhesion, reversible switching between superhydrophobicity and superhydrophilicity, switchable isotropic/anisotropic wetting, slippery surface with tunable wettability, and underwater superaerophobicity/superoleophobicity with tunable adhesion, can be obtained on SMP micro/nanostructures by regulating the surface morphology. Finally, the challenges and future prospects of smart superwetting SMP surfaces are discussed.

A Close Look at Mechanism, Application, and Opportunities of Electrochemiluminescence Microscopy.

Electrochemiluminescence (ECL) is a typical luminescence process triggered by electrochemical reactions. Due to the separated signal types, ECL measurements have some merits of high sensitivity, low background, and simple configuration. Coupled with a microscopy setup, ECL microscopy (ECLM) has the unique characteristics of ECL and is also furnished with spatiotemporal resolution. Thus, many applications have been created, including nanoscale sensing, ECL mechanism deciphering, transient events of single objects, and ECLM crossover methods. In this review, we will overview the development and basic knowledge of ECL and then profile the setup design of ECLM. Through the understanding of these two parts, we will next probe the diverse applications of ECLM, combining the inter-relation with each other. Finally, the outlook discussing the expectations of further progress of ECLM technology.

Rapid Fabrication of Wavelength-Scale Micropores on Metal by Femtosecond MHz Burst Bessel Beam Ablation.

The preparation of the wavelength-scale micropores on metallic surfaces is limited by the high opacity of metal. At present, most micropores reported in the literature are more than 20 µm in diameter, which is not only large in size, but renders them inefficient for processing so that it is difficult to meet the needs of some special fields, such as aerospace, biotechnology, and so on. In this paper, the rapid laser fabrications of the wavelength-scale micropores on various metallic surfaces are achieved through femtosecond MHz burst Bessel beam ablation. Taking advantage of the long-depth focal field of the Bessel beam, high-density micropores with a diameter of 1.3 µm and a depth of 10.5 µm are prepared on metal by MHz burst accumulation; in addition, the rapid fabrication of 2000 micropores can be achieved in 1 s. The guidelines and experimental results illustrate that the formations of the wavelength-scale porous structures are the result of the co-action of the laser-induced periodic surface structure (LIPSS) effect and Bessel beam interference. Porous metal can be used to store lubricant and form a lubricating layer on the metallic surface, thus endowing the metal resistance to various liquids' adhesion. The microporous formation process on metal provides a new physical insight for the rapid preparation of wavelength-scale metallic micropores, and promotes the application of porous metal in the fields of catalysis, gas adsorption, structural templates, and bio-transportation fields.

A temperature-tuned electrochemiluminescence layer for reversibly imaging cell topography.

Investigating electrochemiluminescence (ECL) scenarios under different temperatures is important to expand its imaging scope near an electrode surface. Here, we develop a temperature-tuned ECL layer by recording the evolution of shadow regions of adherent cells. Finite element simulation and experimental results demonstrate that the thickness of the ECL layer (TEL) is reversibly regulated by electrode temperature (T e), so that single cell topography at different heights is imaged. The TEL in two ECL routes shows different regulation ranges with elevated T e, thus providing a flexible approach to adjust the imaging scope within specific heights. In addition, a heated electrode significantly improves the image quality of cell adhesion in heterogeneous electrochemical rate-determined situations. Thus, the contrast in cell regions shows a reversible response to T e. This work provides a new approach to regulate the TEL and is promising for monitoring transient heat generation from biological entities.

A Synergistic Coreactant for Single-Cell Electrochemiluminescence Imaging: Guanine-Rich ssDNA-Loaded High-Index Faceted Gold Nanoflowers.

Developing an electrochemiluminescence (ECL) coreactant to minimize the biotoxicity and to maximize the enhancement factor is critical to single-cell ECL microscopy. Here, we reported a guanine-rich single-stranded DNA (G-ssDNA)-loaded high-index faceted gold nanoflower (Hi-AuNF) as a synergistic coreactant of Ru(bpy)32+ for single-cell ECL imaging. Because of the excellent catalytic performance and huge specific surface area, Hi-AuNF serves as not only an ECL enhancer but also a carrier for G-ssDNA. Guanine in G-ssDNA specifically reacts with Ru(bpy)32+ through a so-called "catalytic route" and thus significantly enhances the ECL signal of Ru(bpy)32+. To endow targeting ability to the synergistic coreactant, an aptamer of carcinoembryonic antigen (CEA) is incorporated into the G-ssDNA to form G-ssDNA-Apt for the recognition of human breast adenocarcinoma cells, which overexpress CEA on the cytomembrane. Accordingly, the ECL imaging of CEA on the cytomembrane was realized by using the highly selective Hi-AuNF@G-ssDNA-Apt as the probe as well as the luminophore of Ru(bpy)32+. Compared with the common coreactant tripropylamine with high toxicity and volatility, the Hi-AuNF@G-ssDNA-Apt is considered as a high-performance and biocompatible coreactant, providing exciting opportunities in single-cell imaging and detection.

Development of a sensitive electrochemical immunosensor using polyaniline functionalized graphene quantum dots for detecting a depression marker.

As a new type of conductive material, polyaniline functionalized graphene quantum dots (PAGD), which were prepared by in-situ polymerization had been used to construct a novel electrochemical immunosensor for early screening of depression markers-heat shock protein 70 (HSP70). Profiting from the huge specific surface area, good bioactivity and excellent structure of PAGD, a variety of heat shock protein 70 (HSP70) was firmly loaded on the surface of PAGD for successful construction of basic electrode (HSP70/PAGD/GCE), which was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), respectively. Due to the HSP70 fixed on the surface of basic electrode and the HSP70 in the samples can competitively combine with the horseradish peroxidase labeled human HSP70 antibody (HRP-Strept-Biotin-Ab). As a result, there is negative correlation between the concentration of HSP70 in samples and the detection signal of the proposed electrochemical immunosensor (HRP-Strept-Biotin-Ab-HSP70/PAGD/GCE) in the test liquid. Under conditions optimized for determining HSP70, wide linearity was obtained in the range of 0.0976-100 ng/mL, with a low detection limit of 0.05 ng/mL at 3σ. Moreover, the proposed electrochemical immunosensors was successfully applied to detect HSP70 in plasma samples, and exhibited good precision, acceptable stability and reproducibility. Therefore, this study provides a novel and convenient method for early clinical screening of depression markers, and also provides a reliable and objective analysis method for the diagnosis of depression at the molecular level.

Porous graphene-black phosphorus nanocomposite modified electrode for detection of leptin.

Leptin is a vital biomarker of non-alcoholic fatty liver (NAFLD), and its evaluation of the concentration level in vivo is of great significance to NAFLD diagnosis. Therefore, it is pressing to develop a method for rapid and sensitive detection of leptin. This paper describes an environmentally friendly and label-free immunosensor based on porous graphene functionalized black phosphorus (PG-BP) composite to detect of leptin. The PG-BP was synthesized via strong coherent coupling between porous graphene (PG) surface plasmons and anisotropic black phosphorus (BP) localized surface plasmons, which made the electrochemical performance of PG and BP synergistic as well as increased the stability and conductive capability of BP material. The PG-BP modified electrodes was further prepared by gold nanoparticles, cysteamine, and glutaraldehyde in turn. Due to the cross-linking effect of glutaraldehyde, anti-leptin can be firmly fixed. These properties of the platform improved the conductive capability of the immunosensor and enhanced the load capacity of the proteins, thereby, the sensitivity of the immunosensor was significantly increased. Under the optimal conditions, the proposed immunosensor exhibited a wide linear range of 0.150-2500 pg/mL with a low detection limit of 0.036 pg/mL. The leptin immunosensor displayed excellent selectivity and anti-interference ability, which could be used for early screening and diagnosis of clinical NALFD.

In Situ Imaging Facet-Induced Spatial Heterogeneity of Electrocatalytic Reaction Activity at the Subparticle Level via Electrochemiluminescence Microscopy.

Investigating catalytic behavior of heterogeneous catalysts, especially at the crystal facets level, is crucial for rational catalyst design in the energy and environmental fields. Here we demonstrate an efficient approach to in situ visualize and analyze the heterogeneity of electrocatalytic activity on different facets at the subparticle level via electrochemiluminescence (ECL) microscopy. ZnO crystals with various exposed facet proportions were synthesized, and the correlation between their electrocatalytic performance toward luminol analogue degradation and the exposed facets is established. It is clearly imaged that the ZnO (002) facet has superior catalytic performance compared to the ZnO (100) facet, which is supported by theoretical computation and electrochemical experiments as the facet-induced heterogeneity of the catalytic effect on oxygen reduction into the key reactant for ECL. Accordingly, the spatial heterogeneity of electrocatalytic activity at different facets on one particle is visualized for the first time. The realization of subparticle ECL imaging and kinetic analysis could provide a special approach to visualize facet-induced spatial heterogeneity of catalytic behavior and valuable information for the catalysis study and analysis.

A novel electrochemical immunosensor based on PG for early screening of depression markers-heat shock protein 70.

In this study, a novel electrochemical immunosensor for early screening of depression markers-heat shock protein 70 (HSP70) was successfully developed based on the porous graphene (PG) with huge specific surface area and excellent structure. Benefiting from the strong adsorption and good bioactivity of PG which was initially prepared via a simple pyrolysis process, a variety of heat shock protein70 (HSP70) can be firmly loaded on the PG to construct the basic electrode (HSP70/PG/GCE)，which was characterized by the cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), respectively. Due to the HSP70 fixed on the surface of basic electrode and the HSP70 in the samples can competitively combine with the horseradish peroxidase labeled human HSP 70 antibody (HRP-Strept-Biotin-Ab). As a result, it presented a negative correlation between the concentration of HSP70 in samples and the detection signal of the proposed electrochemical immunosensor (HRP-Strept-Biotin-Ab-HSP70/PG/GCE) in the test liquid. The application of PG with excellent electrical conductivity in construction of immunosensor remarkably improved the sensitivity of the immunosensor for detection of HSP70. The proposed immunosensor demonstrated a wide linear range of 0.0448 ~ 100 ng/mL with a low detection limit of 0.02 ng/mL at 3σ. Moreover, the proposed immunosensor could be applied for the sensitive and efficient detection of HSP70 in real samples with good precision, acceptable stability, reproducibility and satisfactory results. Therefore, the HSP70 immunosensor provides a novel and convenient method for early clinical screening of depression markers-heat shock protein 70.

Direct electrochemistry and electrocatalysis of lobetyolin via magnetic functionalized reduced graphene oxide film fabricated electrochemical sensor.

A novel lobetyolin electrochemical sensor based on a magnetic functionalized reduced graphene oxide/Nafion nanohybrid film has been introduced in this work. The magnetic functionalized reduced graphene oxide was characterized by fourier transform infrared spectroscopy, atomic force microscope, X-ray diffraction, transmission electron microscopy and thermogravimetric analysis. The scanning electron microscopy characterized the morphology and microstructure of the prepared sensors, and the electrochemical effective surface areas of the prepared sensors were also calculated by chronocoulometry method. The electrochemical behavior of lobetyolin on the magnetic functionalized reduced graphene oxide/Nafion nanohybrid modified glassy carbon electrode was investigated by cyclic voltammetry and differential pulse voltammetry in a phosphate buffer solution of pH6.0. The electron-transfer coefficient (α), electron transfer number (n), and electrode reaction rate constant (Κs) were calculated as 0.78, 0.73, and 4.63s-1, respectively. Under the optimized conditions, the sensor based on magnetic functionalized reduced graphene oxide/Nafion showed a linear voltammetric response to the lobetyolin concentration at 1.0×10-7 to 1.0×10-4mol/L with detection limit (S/N=3)of 4.3×10-8mol/L. The proposed sensor also displayed acceptable reproducibility, long-term stability, and high selectivity, and performs well for analysis of lobetyolin in real samples. The voltammetric sensor was successfully applied to detect lobetyolin in Codonopsis pilosula with recovery values in the range of 96.12%-102.66%.