Deciphering the Dynamic Assembling-Disassembling of Small Molecules on Solid/Liquid Interfaces within Microchannels by Pulsed Streaming Potential Measurement.

Assembling small molecules at liquid/solid interfaces is relatively common and contributes to many unique properties of the interface. However, such an assembling process can be dynamic depending on the concentration of the molecule and the properties of the solid and liquid themselves, which poses serious challenges on the accurate evaluation of the assembling processes. Herein, we report a convenient way for in situ and real-time monitoring of assembling-disassembling of small-molecule surfactants on the surface of microchannels using pulsed streaming potential (SP) measurement based on the variation of surface charge. With this technique, five distinctive kinetic regimes, each responsible for a characteristic molecular behavior, can be differentiated during a typical assembling-disassembling cycle. Significant difference of the assembling-disassembling process was clearly reflected for surfactants with hydrophobic tails of only a two -CH2- difference (C16TAB/C18TAB and D10DAB/D12DAB). The relative SP (Er) value is positively correlated with the molecular weight at a concentration of 0.1 mM for the same kinds of surfactants. Moreover, the assembling kinetics of D10DAB exhibits an "overshoot effect" at high concentration, which means morphology adjustment. The consequences of such assembling/disassembling of these molecules for electrophoretic separation, protein immobilization, and photocatalysis in a microchannel were investigated through dynamic characterization, which proves its potential as a tool for dynamic solid/liquid interface characterization.

Dual-Stimuli Regulation of DNAzyme Cleavage Reaction by Coordination-Driven Nanoprobes for Cancer Cell Imaging.

Endowing current artificial chemical reactions (ACRs) with high specificity and intricate activation capabilities is crucial for expanding their applications in accurate bioimaging within living cells. However, most of the reported ACR-based evaluations relied on either single biomarker stimuli or dual activators without obvious biological relevance, still limiting their accuracy and fidelity. Herein, taking the metal-ion-dependent DNAzyme cleavage reaction as a model ACR, two regulators, glutathione (GSH) and telomerase (TE) activated DNAzyme cleavage reactions, were exploited for precise discrimination of cancerous cells from normal cells. DNA probe was self-assembled into the ZIF-90 nanoparticle framework to construct coordination-driven nanoprobes. This approach enhances the stability and specificity of tumor imaging by utilizing biomarkers associated with rapid tumor proliferation and those commonly overexpressed in tumors. In conclusion, the research not only paves the way for new perspectives in cell biology and pathology studies but also lays a solid foundation for the advancement of biomedical imaging and disease diagnostic technologies.

Monolithic 3D nanoelectrospray emitters based on a continuous fluid-assisted etching strategy for glass droplet microfluidic chip-mass spectrometry.

Glass microfluidic chips are suitable for coupling with mass spectrometry (MS) due to their flexible design, optical transparency and resistance to organic reagents. However, due to the high hardness and brittleness of glass, there is a lack of simple and feasible technology to manufacture a monolithic nanospray ionization (nESI) emitter on a glass microchip, which hinders its coupling with mass spectrometry. Here, a continuous fluid-assisted etching strategy is proposed to fabricate monolithic three-dimensional (3D) nESI emitters integrated into glass microchips. A continuous fluid of methanol is adopted to protect the inner wall of the channels and the bonding interface of the glass microfluidic chip from being wet-etched, forming sharp 3D nESI emitters. The fabricated 3D nESI emitter can form a stable electrospray plume, resulting in consistent nESI detection of acetylcholine with an RSD of 4.5% within 10 min. The fabricated 3D emitter is integrated on a glass microfluidic chip designed with a T-junction droplet generator, which can realize efficient analysis of acetylcholine in picoliter-volume droplets by nESI-MS. Stability testing of over 20 000 droplets detected by the established system resulted in an RSD of 9.1% over approximately 180 min. The detection of ten neurochemicals in rat cerebrospinal fluid droplets is achieved. The established glass droplet microfluidic chip-MS system exhibits potential for broad applications such as in vivo neurochemical monitoring and single-cell analysis in the future.

Label-Free Direct Identification of MicroRNAs Based on a Narrow Constant-Inner-Diameter Emitter Mass Spectrometry Analysis.

MicroRNAs (miRNAs) are a class of endogenous noncoding small RNAs that play important roles in various biological processes and diseases. Direct determination of miRNAs is a cost-efficient and accurate method for analysis. Herein, we established a novel method for the analysis of miRNAs based on a narrow constant-inner-diameter mass spectrometry emitter. We utilized the gravity-assisted sleeving etching method to prepare a constant-inner-diameter mass spectrometry emitter with a capillary inner diameter of 5.5 µm, coupled it with a high-voltage power supply and a high-resolution mass spectrometer, and used it for miRNA direct detection. The method showed high sensitivity and reproducibility for the analysis of four miRNAs, with a limit of detection of 100 nmol/L (170 amol) for the Hsa-miR-1290 analysis. Compared with commercial ion sources, our method achieved higher sensitivity for miRNA detection. In addition, we analyzed the total miRNAs in the A549 cells. The result indicated that both spiked and endogenous miRNAs could be quantified with high accuracy. As a result, this method offers a promising platform for highly sensitive and accurate miRNA analysis. Furthermore, this approach can be extended to the analysis of other small oligonucleotides and holds the potential for studying clinical samples and facilitating disease diagnosis.

Ultra-Large Two-Dimensional Metal Nanowire Networks by Microfluidic Laminar Flow Synthesis for Formic Acid Electrooxidation.

Despite the great research interest in two-dimensional metal nanowire networks (2D MNWNs) due to their large specific surface area and abundance of unsaturated coordination atoms, their controllable synthesis still remains a significant challenge. Herein, a microfluidics laminar flow-based approach is developed, enabling the facile preparation of large-scale 2D structures with diverse alloy compositions, such as PtBi, AuBi, PdBi, PtPdBi, and PtAuCu alloys. Remarkably, these 2D MNWNs can reach sizes up to submillimeter scale (~220 µm), which is significantly larger than the evolution from the 1D or 3D counterparts that typically measure only tens of nanometers. The PdBi 2D MNWNs affords the highest specific activity for formic acid (2669.1 mA mg-1) among current unsupported catalysts, which is 103.5 times higher than Pt-black, respectively. Furthermore, in situ Fourier transform infrared (FTIR) experiments provide comprehensive evidence that PdBi 2D MNWNs catalysts can effectively prevent CO* poisoning, resulting in exceptional activity and stability for the oxidation of formic acid.

Enhanced recognition of G-quadruplex DNA oxidative damage based on DNA-mediated charge transfer.

G-quadruplex (G4) DNA is present in human telomere oligonucleotide sequences. Oxidative damage to telomeric DNA accelerates telomere shortening, which is strongly associated with aging and cancer. Most of the current analyses on oxidative DNA damage are based on ds-DNA. Here, we developed a electrochemiluminescence (ECL) probe for enhanced recognition of oxidative damage in G4-DNA based on DNA-mediated charge transfer (CT), which could specifically recognize damaged sites depending on the position of 8-oxoguanine (8-oxoG). First, a uniform G4-DNA monolayer interface was fabricated; the G4-DNA mediated CT properties were examined using an iridium(III) complex [Ir(ppy)2(pip)]PF6 stacked with G4-DNA as an indicator. The results showed that G4-DNA with 8-oxoG attenuated DNA CT. The topological effects of oxidative damage at different sites of G4-DNA and their effects on DNA CT were revealed. The sensing platform was also used for the sensitive and quantitative detection of 8-oxoG in G4-DNA, with a detection limit of 28.9 fmol. Overall, these findings present a sensitive platform to study G4-DNA structural and stability changes caused by oxidative damage as well as the specific and quantitative detection of oxidation sites. The different damage sites in the G-quadruplex could provide detailed clues for understanding the function of G4-associated telomere functional enzymes.

Assessing the Redox Toxicity of 2D Nanosheets Based on Their Redox Effect on Cytochrome c in Microchannels.

2D nanosheets (NSs) have been widely used in drug-related applications. However, a comprehensive investigation into the cytotoxicity mechanism linked to the redox activity is lacking. In this study, with cytochrome c (Cyt c) as the model biospecies, the cytotoxicity of 2D NSs was evaluated systematically based on their redox effect with microfluidic techniques. The interface interaction, dissolution, and redox effect of 2D NSs on Cyt c were monitored with pulsed streaming potential (SP) measurement and capillary electrophoresis (CE). The relationship between the redox activity of 2D NSs and the function of Cyt c was evaluated in vitro with Hela cells. The results indicated that the dissolution and redox activity of 2D NSs can be simultaneously monitored with CE under weak interface interactions and at low sample volumes. Both WS2 NSs and MoS2 NSs can reduce Cyt c without significant dissolution, with reduction rates measured at 6.24 × 10-5 M for WS2 NSs and 3.76 × 10-5 M for MoS2 NSs. Furthermore, exposure to 2D NSs exhibited heightened reducibility, which prompted more pronounced alterations associated with Cyt c dysfunction, encompassing ATP synthesis, modifications in mitochondrial membrane potential, and increased reactive oxygen species production. These observations suggest a positive correlation between the redox activity of 2D NSs and their redox toxicity in Hela cells. These findings provide valuable insight into the redox properties of 2D NSs regarding cytotoxicity and offer the possibility to modify the 2D NSs to reduce their redox toxicity for clinical applications.

Kinetically Controlled Nucleation Enabled by Tunable Microfluidic Mixing for the Synthesis of Dendritic Au@Pt Core/Shell Nanomaterials.

The nucleation stage plays a decisive role in determining nanocrystal morphology and properties; hence, the ability to regulate nucleation is critical for achieving high-level control. Herein, glass microfluidic chips with S-shaped mixing units are designed for the synthesis of Au@Pt core/shell materials. The use of hydrodynamics to tune the nucleation kinetics is explored by varying the number of mixing units. Dendritic Au@Pt core/shell nanomaterials are controllably synthesized and a formation mechanism is proposed. As-synthesized Au@Pt exhibited excellent ethanol oxidation activity under alkaline conditions (8.4 times that of commercial Pt/C). This approach is also successfully applied to the synthesize of Au@Pd core/shell nanomaterials, thus demonstrating its generality.

Synthesis of Multiple Emission Carbon Dots from Dihydroxybenzoic Acid via Decarboxylation Process.

Carbon dots (CDs), as a new zero-dimensional carbon-based nanomaterial with desirable optical properties, exhibit great potential for many application fields. However, the preparation technique of multiple emission CDs with high yield is difficult and complex. Therefore, exploring the large-scale and straightforward synthesis of multicolor CDs from a simple carbon source is necessary. In this work, the solvent-free method prepares a series of multicolor emission CDs from dihydroxybenzoic acid (DHBA). The maximum emission wavelengths are 408, 445, 553, 580, and 610 nm, respectively, covering the visible light region. The 2,4- and 2,6-CDs possess the longer emission wavelength caused by the 2,4-, and 2,6-DHBA easily undergo decarboxylation to form the larger sp2 domain graphitized structure. These CDs incorporated with g-C3N4 can significantly improve the photocatalytic water-splitting hydrogen production rate by extending the visible light absorption and enhancing the charge separation efficiency. The long-wavelength emission CDs can further enhance photocatalytic activity primarily by improving visible light absorption efficiency.

Sensitive electrochemical measurement of nitric oxide released from living cells based on dealloyed PtBi alloy nanoparticles.

Nitric oxide (NO), as a vital signaling molecule related to different physiological and pathological processes in living systems, is closely associated with cancer and cardiovascular disease. However, the detection of NO in real-time remains a difficulty. Here, PtBi alloy nanoparticles (NPs) were synthesized, dealloyed, and then fabricated to NP-based electrodes for the electrochemical detection of NO. Transmission electron microscopy (TEM), small-angle X-ray scattering (SAXS), and nitrogen physical adsorption/desorption show that dealloyed PtBi alloy nanoparticles (dPtBi NPs) have a porous nanostructure. Electrochemical impedance spectroscopy and cyclic voltammetry results exhibit that the dPtBi NP electrode possesses unique electrocatalytic features such as low charge transfer resistance and large electrochemically active surface area, which lead to its excellent NO electrochemical sensing performance. Owing to the higher density of catalytical active sites formed PtBi bimetallic interface, the dPtBi NP electrode displays superior electrocatalytic activity toward the oxidation of NO with a peak potential at 0.74 V vs. SCE. The dPtBi NP electrode shows a wide dynamic range (0.09-31.5 µM) and a low detection limit of 1 nM (3σ/k) as well as high sensitivity (130 and 36.5 µA µM-1 cm-2). Moreover, the developed dPtBi NP-based electrochemical sensor also exhibited good reproducibility (RSD 5.7%) and repeatability (RSD 3.4%). The electrochemical sensor was successfully used for the sensitive detection of NO produced by live cells. This study indicates a highly effective approach for regulating the composition and nanostructures of metal alloy nanomaterials, which might provide new technical insights for developing high-performance NO-sensitive systems, and have important implications in enabling real-time detection of NO produced by live cells.

Formation of Protein Nanoparticles in Microdroplet Flow Reactors.

Nanoparticles are increasingly being used for biological applications, such as drug delivery and gene transfection. Different biological and bioinspired building blocks have been used for generating such particles, including lipids and synthetic polymers. Proteins are an attractive class of material for such applications due to their excellent biocompatibility, low immunogenicity, and self-assembly characteristics. Stable, controllable, and homogeneous formation of protein nanoparticles, which is key to successfully delivering cargo intracellularly, has been challenging to achieve using conventional methods. In order to address this issue, we employed droplet microfluidics and utilized the characteristic of rapid and continuous mixing within microdroplets in order to produce highly monodisperse protein nanoparticles. We exploit the naturally occurring vortex flows within microdroplets to prevent nanoparticle aggregation following nucleation, resulting in systematic control over the particle size and monodispersity. Through combination of simulation and experiment, we find that the internal vortex velocity within microdroplets determines the uniformity of the protein nanoparticles, and by varying parameters such as protein concentration and flow rates, we are able to finely tune nanoparticle dimensional properties. Finally, we show that our nanoparticles are highly biocompatible with HEK-293 cells, and through confocal microscopy, we determine that the nanoparticles fully enter into the cell with almost all cells containing them. Due to the high throughput of the method of production and the level of control afforded, we believe that the approach described in this study for generating monodisperse protein-based nanoparticles has the potential for intracellular drug delivery or for gene transfection in the future.

Quantitative Analysis of Nanoplastics in Single Cells by Subcellular Chromatography.

The accumulation and spatial distribution of intracellular nanoplastic particles provide useful information about their spatiotemporal toxicological effects mediated by the physicochemical parameters of nanoplastics in living cells. In this study, a sample injection-transfer method was designed with an accuracy of up to femtoliters to attoliters to match the volume required for ultranarrow-bore open-tubular liquid chromatography. The separation and concentration quantification of mixed polystyrenes in different regions in living cells were achieved by directly transferring picoliter/femtoliter volumes of intracellular cytoplasm to an ultranarrow-bore open-tubular chromatographic column. The measurement of pollutant concentration in different areas of a small-volume target (single cell) was realized. This method is expected to be used in the qualitative and quantitative analyses of complex, mixed, and label-free nanoplastics (a few nm in size) in the subregions of living cells.

Biomimetic Nanocomposites Camouflaged with Hybrid Cell Membranes for Accurate Therapy of Early-Stage Glioma.

Glioma features high fatality rate and short survival time of patients due to its fast growth speed and high invasiveness, hence timely treatment of early-stage glioma is extremely important. However, the blood brain barrier (BBB) severely prevents therapeutic agents from entering the brain; meanwhile, the non-targeted distribution of agents always causes side effects to vulnerable cerebral tissues. Therefore, delivery systems that possess both BBB penetrability and precise glioma targeting ability are keenly desired. We herein proposed a hybrid cell membrane (HM) camouflage strategy to construct therapeutic nanocomposites, in which HM consisting of brain metastatic breast cancer cell membrane and glioma cell membrane was prepared with a simple membrane fusion pathway. By coating HM onto drug-loaded nanoparticles, the as-obtained biomimetic therapeutic agent (termed HMGINPs) inherited satisfying BBB penetrability and homologous glioma targeting ability simultaneously from the two source cells. HMGINPs exhibited good biocompatibility and superior therapeutic efficacy towards early-stage glioma.

Dealloying of Pt1Bi2 intermetallic toward optimization of electrocatalysis on a Bi-continuous nanoporous core-shell structure.

Noble metal nanoporous materials hold great potential in the field of catalysis, owing to their high open structures and numerous low coordination surface sites. However, the formation of porous nanoparticles is restricted by particle size. Herein, we utilized a Pt1Bi2 intermetallic nanocatalyst to develop a dealloying approach for preparing nanoparticles with a bi-continuous porous and core-shell structure and proposed a mechanism for the formation of pores. The particle size used to form the porous structure can be <10 nm, which enhances the nanocatalyst's performance for the oxygen reduction reaction (ORR). This study provides a new understanding of the formation of porous materials via a dealloying approach.

Single-Cell Metabolomics-Based Strategy for Studying the Mechanisms of Drug Action.

Studying the mechanisms of drug antitumor activity at the single-cell level can provide information about the responses of cell subpopulations to drug therapy, which is essential for the accurate treatment of cancer. Due to the small size of single cells and the low contents of metabolites, metabolomics-based approaches to studying the mechanisms of drug action at the single-cell level are lacking. Herein, we develop a label-free platform for studying the mechanisms of drug action based on single-cell metabolomics (sMDA-scM) by integrating intact living-cell electro-launching ionization mass spectrometry (ILCEI-MS) with metabolomics analysis. Using this platform, we reveal that non-small-cell lung cancer (NSCLC) cells treated by gefitinib can be clustered into two cell subpopulations with different metabolic responses. The glutathione metabolic pathway of the subpopulation containing 14.4% of the cells is not significantly affected by gefitinib, exhibiting certain resistance characteristics. The presence of these cells masked the judgment of whether cysteine and methionine metabolic pathway was remarkably influenced in the analysis of overall average results, revealing the heterogeneity of the response of single NSCLC cells to gefitinib treatment. The findings provide a basis for evaluating the early therapeutic effects of clinical medicines and insights for overcoming drug resistance in NSCLC subpopulations.

An isolated single-particle-based SECM tip interface for single-cell NO sensing.

As a key factor in cellular metabolic processes, nitric oxide (NO) is a challenging target for in situ real-time monitoring due to its transient property and short diffusion distance. Scanning electrochemical microscopy (SECM) has unique advantages in single-cell analysis, which can obtain the electrochemical current by scanning the cell surface with a tip microelectrode. In particular, it can further improved the electrochemical response by enhancing the interface properties of its tip. Here, an interface design strategy based on platinum single nanoparticle (Pt NP) was developed, and fluorinated self-assembled monolayers (SAMs) were used to further improve its performance. This modified tip was used as an SECM probe for NO concentration monitoring and morphological imaging of single MCF-7 cells. It has the high sensitivity (164.7 µA/µM·cm2) and good selectivity for NO detection, which benefits from the efficient catalytic properties of Pt NPs and high mass transport and hydrophobic antifouling properties of the interface. Notably, it shows a superior performance in detecting the fluctuation of NO released by a single MCF-7-cell under the stimulation of cadmium (Cd), which demonstrates a promising method for using a single-particle-based tip in SECM applications.

The Formation Process and Mechanism of Carbon Dots Prepared from Aromatic Compounds as Precursors: A Review.

Fluorescent carbon dots are a novel type of nanomaterial. Due to their excellent optical properties, they have extensive application prospects in many fields. Studying the formation process and fluorescence mechanism of CDs will assist scientists in understanding the synthesis of CDs and guide more profound applications. Due to their conjugated structures, aromatic compounds have been continuously used to synthesize CDs, with emissions ranging from blue to NIR. There is a lack of a systematic summary of the formation process and fluorescence mechanism of aromatic precursors to form CDs. In this review, the formation process of CDs is first categorized into three main classes according to the precursor types of aromatic compounds: amines, phenols, and polycyclics. And then, the fluorescence mechanism of CDs synthesized from aromatic compounds is summarized. The challenges and prospects are proposed in the last section.

Clusterbody Enables Flow Sorting-Assisted Single-Cell Mass Spectrometry Analysis for Identifying Reversal Agent of Chemoresistance.

Identifying effective reversal agents overcoming multidrug resistance with causal mechanisms from an efflux pump protein is of vital importance for enhanced tumor chemotherapy in clinic. To achieve this end, we construct a metal cluster-based probe, named clusterbody, to develop flow sorting-assisted single-cell mass spectrometry analysis. This clusterbody synthesized by biomimetic mineralization possesses an antibody-like property to selectively recognize an efflux pump protein. The intrinsic red fluorescence emission of the clusterbody facilitates fluorescence-activated high-throughput cell sorting of subpopulations with different multidrug resistance levels. Furthermore, based on the accurate formula of the clusterbody, the corresponding protein abundance at the single-cell level is determined through detecting gold content via precise signal amplification by laser ablation inductively coupled plasma mass spectrometry. Therefore, the effect of reversal agent treatment overcoming multidrug resistance is evaluated in a quantitative manner. This work opens a new avenue to identify reversal agents, shedding light on developing combined or synergetic tumor therapy.

Spontaneous Phase Segregation Enabling Clogging Aversion in Continuous Flow Microfluidic Synthesis of Nanocrystals Supported on Reduced Graphene Oxide.

Eliminating clogging in capillary tube reactors is critical but challenging for enabling continuous-flow microfluidic synthesis of nanoparticles. Creating immiscible segments in a microfluidic flow is a promising approach to maintaining a continuous flow in the microfluidic channel because the segments with low surface energy do not adsorb onto the internal wall of the microchannel. Herein we report the spontaneous self-agglomeration of reduced graphene oxide (rGO) nanosheets in polyol flow, which arises because the reduction of graphene oxide (GO) nanosheets by hot polyol changes the nanosheets from hydrophilic to hydrophobic. The agglomerated rGO nanosheets form immiscible solid segments in the polyol flow, realizing the liquid-solid segmented flow to enable clogging aversion in continuous-flow microfluidic synthesis. Simultaneous reduction of precursor species in hot polyol deposits nanocrystals uniformly dispersed on the rGO nanosheets even without surfactant. Cuprous oxide (Cu2O) nanocubes of varying edge lengths and ultrafine metal nanoparticles of platinum (Pt) and palladium (Pd) dispersed on rGO nanosheets have been continuously synthesized using the liquid-solid segmented flow microfluidic method, shedding light on the promise of microfluidic reactors in synthesizing functional nanomaterials.

A dual-readout paper-based analytical device for the simultaneous determination of hexavalent Cr and total Cr.

A paper-based analytical device (PAD) is presented with colorimetric/electrochemical dual readouts for the simultaneous sensing of total chromium (Cr) and hexavalent chromium (Cr(VI)). This device consists of a homemade three-electrode system and a patterned paper chip, integrating multiple functions including electrochemical detection, fluid driving, online oxidation, and colorimetric detection. The fiberglass filter paper with a hydrophilic microchannel was used to achieve self-driving fluidics without external equipment. One end of the microchannel was integrated with a homemade three-electrode system to achieve sample loading and electrochemical detection. The middle region on the microchannel was modified with oxidizing reagents to perform online pretreatment, and the yield of Cr(III) oxidation can reach 97.9%, ensuring reliable colorimetric detection of total Cr at another end of the microchannel modified with chromogenic agents. With this device, the signals of Cr(VI) (the signal peak at 0.29 V vs. Ag/AgCl) and total Cr can be obtained in one single injection. After optimization, the limit of detection (LOD) of Cr(VI) and total Cr were 0.01 mg L-1 and 0.06 mg L-1 and the linear ranges were 0.05-3.0 mg L-1 and 0.2-3.0 mg L-1, respectively. The relative standard deviations (RSD) of the electrochemical testing of Cr(VI) results were in a range 1.3%-8.7% (n = 3), and the RSD values of the colorimetric testing of total Cr were between 0.7-9.2% (n = 3). The device's reliability was demonstrated by performing the practical speciation of Cr in tap water, river water, and electroplating wastewater while the recoveries obtained using the present method were in the range 93.5-106%. Overall, the proposed device provides high application prospect in the on-site rapid Cr speciation.