Monitoring the response of a model protocell to dye and surfactant molecules through second harmonic generation and fluorescence imaging.

Probing the interaction between molecules and protocells is crucial for understanding the passive transport of functional molecules in and out of artificial and real cells. Second-harmonic generation (SHG) has been proven to be a powerful method for analyzing the adsorption and cross-membrane transport of molecules on lipid bilayers. In this study, we used SHG and two-photon fluorescence (TPF) imaging to study the interaction of charged dye molecules (D289) with a lipid vesicle. Unexpectedly, it was observed that the transport of D289 at a relatively high concentration is not as efficient as that at a lower dye concentration. Periodic shrinking of the model protocell and discharging of D289 out from the vesicle were revealed by combined analyses of SHG and TPF images. The response of the vesicle to a surfactant was also analyzed with D289 as a probe. This work demonstrates that the combined SHG and TPF imaging method is a unique approach that can provide detailed information on the interaction of molecules and lipids (both morphology and molecular kinetics). Determining these subtle interfacial kinetics in molecules is important for understanding the mechanism of many biophysical processes occurring on lipids.

Probing the structural evolution on the surface of cardiolipin vesicles with an amphiphilic second harmonic generation and fluorescence probe.

Investigating the influence of the ambient chemical environment on molecular behaviors in liposomes is crucial for understanding and manipulating cellular vitality as well as the capabilities of lipid drug carriers in various environments. Here, we designed and synthesized a second harmonic generation (SHG) and fluorescence probe molecule called Pyr-Py+-N+ (PPN), which possesses membrane-targeting capability. We employed PPN to investigate the response of lipid vesicles composed of cardiolipin to the presence of exogenous salt. The kinetic behaviors, including the adsorption and embedding of PPN on the surface of small unilamellar vesicles (SUVs) composed of cardiolipin, were analyzed. The response of the SUVs to the addition of NaCl was also monitored. A rapid decrease in vesicle size can be evidenced through the rapid drop in SHG emission originating from PPN located on the vesicle surface.

Single-Atom Catalysts and Dual-Atom Catalysts for CO2 Electroreduction: Competition or Cooperation?

Developing highly active and selective electrocatalysts for electrochemical reduction of CO2 can reduce environmental pollution and mitigation of greenhouse gas emission. Owing to maximal atomic utilization, the atomically dispersed catalysts are broadly adopted in CO2 reduction reaction (CO2 RR). Dual-atom catalysts (DACs), with more flexible active sites, distinct electronic structures, and synergetic interatomic interactions compared to single-atom catalysts (SACs), may have great potential to enhance catalytic performance. Nevertheless, most of the existing electrocatalysts have low activity and selectivity due to their high energy barrier. Herein, 15 electrocatalysts are explored with noble metallic (Cu, Ag, and Au) active sites embedded in metal-organic hybrids (MOHs) for high-performance CO2 RR and studied the relationship between SACs and DACs by first-principles calculation. The results indicated that the DACs have excellent electrocatalytic performance, and the moderate interaction between the single- and dual-atomic center can improve catalytic activity in CO2 RR. Four among the 15 catalysts, including (CuAu), (CuCu), Cu(CuCu), and Cu(CuAu) MOHs inherited a capability of suppressing the competitive hydrogen evolution reaction with favorable CO overpotential. This work not only reveals outstanding candidates for MOHs-based dual-atom CO2 RR electrocatalysts but also provides new theoretical insights into rationally designing 2D metallic electrocatalysts.

F···Br and F···S Heterohalogen-Bond-Directed 2D Self-Assemblies of a Benzothiadiazole Derivative.

Halogen bonding (XB) is of great importance in fabricating a two-dimensional (2D) self-assembly for its adaptive directionality. However, the XBs involving fluorine (F) have barely been studied due to the absence of an σ-hole on F. Here, 2D self-assemblies of a F-substituted 4,7-bis(5-bromo-4-dodecylthiophen-2-yl)-5,6-difluorobenzo[c][1,2,5]thiadiazole (BTZ-BrF) molecule on graphite were investigated using scanning tunneling microscopy (STM) and density functional theory (DFT) calculations. STM experiments revealed that the 2D patterns of BTZ-BrF had a clear solvent and concentration dependence, showing a frame-like pattern in aliphatic acid and aliphatic hydrocarbon solvents at high concentrations. At lower concentrations, a bamboo-like and a wave-like pattern were observed in aliphatic acid, whereas small frame-like and large ladder-like domains at high solution concentrations in aliphatic hydrocarbon were observed. As the concentration further decreased, two linear patterns were observed. DFT calculations suggested that the hetero-XBs of F···Br, F···S, Br···S, and Br···N, the homo-XBs of type-II Br···Br, and the S···S interactions synergistically directed and stabilized the polymorphic 2D architectures. This understanding of intermolecular XBs during the molecular assembly at the molecular level may shed light on the ongoing efforts of regulating nanostructures of multifunctional organics.

Theoretically Revealed and Experimentally Demonstrated Synergistic Electronic Interaction of CoFe Dual-Metal Sites on N-doped Carbon for Boosting Both Oxygen Reduction and Evolution Reactions.

Heteronuclear double-atom catalysts, unlike single atom catalysts, may change the charge density of active metal sites by introducing another metal single atom, thereby modifying the adsorption energies of reaction intermediates and increasing the catalytic activities. First, density functional theory calculations are used to figure out the best combination by modeling two transition-metal atoms from Fe, Co, and Ni onto N-doped graphene. Generally, Fe and Co sites are highly active for the oxygen reduction reaction (ORR) and the oxygen evolution reaction (OER), respectively. The combination of Co and Fe to form CoFe-N-C not only further improves the Fe's ORR and Co's OER activities but also greatly enhances the Co site's ORR and Fe site's OER activities. Then, we synthesize the CoFe-N-C by a two-step pyrolysis process and find that the CoFe-N-C exhibits exceptional ORR and OER electrocatalytic activities in alkaline media, significantly superior to Fe-N-C and Co-N-C and even commercial catalysts.

The Transport of Charged Molecules across Three Lipid Membranes Investigated with Second Harmonic Generation.

Subtle variations in the structure and composition of lipid membranes can have a profound impact on their transport of functional molecules and relevant cell functions. Here, we present a comparison of the permeability of bilayers composed of three lipids: cardiolipin, DOPG (1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol), and POPG (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol)). The adsorption and cross-membrane transport of a charged molecule, D289 (4-(4-diethylaminostyry)-1-methyl-pyridinium iodide), on vesicles composed of the three lipids were monitored by second harmonic generation (SHG) scattering from the vesicle surface. It is revealed that structural mismatching between the saturated and unsaturated alkane chains in POPG leads to relatively loose packing structure in the lipid bilayers, thus providing better permeability compared to unsaturated lipid bilayers (DOPG). This mismatching also weakens the efficiency of cholesterol in rigidifying the lipid bilayers. It is also revealed that the bilayer structure is somewhat disturbed by the surface curvature in small unilamellar vesicles (SUVs) composed of POPG and the conical structured cardiolipin. Such subtle information on the relationship between the lipid structure and the molecular transport capability of the bilayers may provide clues for drug development and other medical and biological studies.

Ultrarapid Nanomanufacturing of High-Quality Bimetallic Anode Library toward Stable Potassium-Ion Storage.

Bimetallic alloy nanomaterials are promising anode materials for potassium-ion batteries (KIBs) due to their high electrochemical performance. The most well-adopted fabrication method for bimetallic alloy nanomaterials is tube furnace annealing (TFA) synthesis, which can hardly satisfy the trade-off among granularity, dispersity and grain coarsening due to mutual constraints. Herein, we report a facile, scalable and ultrafast high-temperature radiation (HTR) method for the fabrication of a library of ultrafine bimetallic alloys with narrow size distribution (≈10-20 nm), uniform dispersion and high loading. The metal-anchor containing heteroatoms (i.e., O and N), ultrarapid heating/cooling rate (≈103  K s-1 ) and super-short heating duration (several seconds) synergistically contribute to the successful synthesis of small-sized alloy anodes. As a proof-of-concept demonstration, the as-prepared BiSb-HTR anode shows ultrahigh stability indicated by negligible degradation after 800 cycles. The in situ X-ray diffraction reveals the K+ storage mechanism of BiSb-HTR. This study can shed light on the new, rapid and scalable nanomanufacturing of high-quality bimetallic alloys toward extended applications of energy storage, energy conversion and electrocatalysis.

Exploiting the Synergistic Electronic Interaction between Pt-Skin Wrapped Intermetallic PtCo Nanoparticles and Co-N-C Support for Efficient ORR/EOR Electrocatalysis in a Direct Ethanol Fuel Cell.

The development of low-Pt catalysts with high activity and durability is critical for fuel cells. Here, Pt-skin wrapped sub-5 nm PtCo intermetallic nanoparticles are successfully mounted on single atom Co-N-C support by exploiting the barrier effect of Co-anchor. According to a collaborative experimental and computational investigation, the increased oxygen reduction reaction activity of PtCo/Co-N-C arises from the direct electron transfer from PtCo to Co-N-C, and the resulting optimal d-band center of Pt. Owing to such unique electronic structure interaction and synergistic effect, the specific and mass activities of PtCo/Co-N-C are up to 4.20 mA cm-2 and 2.71 A mgPt-1 , respectively, with barely degraded stability after 40 000 CV cycles. The PtCo/Co-N-C also exhibits outstanding activity as an ethanol electrocatalyst. This work shows a new and effective route to boost the overall efficiency of direct ethanol fuel cells in acidic media by integrating intermetallic low-Pt alloys and single atom carbon support.

Understanding the different cross-membrane transport kinetics of two charged molecules on the DOPG lipid surface with second harmonic generation and MD simulation.

A clear physical picture of the dynamic behavior of molecules on the surface of the lipid membrane is highly desired and has attracted great attention from researchers. In this study, a step forward in this direction based on previous studies was presented with second harmonic generation (SHG) and molecular dynamic (MD) simulation. Specifically, details on the orientation flipping and cross-membrane transport of two charged molecules, 4-(4-diethylaminostyry)-1-methyl-pyridinium iodide (D289) and malachite green (MG), on the surface of 2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG) lipids were presented. Firstly, the orientation flipping of the two molecules on the surface of lipids before their cross-membrane transport was confirmed by the MD simulation. Then, the concentration dependent rate of the cross membrane transport for MG/D289 was analyzed. It was found that a simplified model could satisfactorily interpret the faster cross-membrane transport of MG under higher bulk concentrations. A different concentration dependent dynamics was observed with D289 and the reason behind it was also discussed. With this investigation, the surface structures and dynamics of D289 and MG on the DOPG lipid surface were clearly presented.

Revealing the mechanisms of vesicle formation with multiple spectral methods.

The investigation of the self-assembly of amphiphilic molecules and the formation of micelles/vesicles has attracted significant attention. However, in situ and real-time methods for such studies are rare. Here, a surface-sensitive second harmonic generation (SHG) technique was applied to study the formation of vesicles in solutions of an anti-cancer drug, doxorubicin (DOX), and a generally used surfactant (sodium bis (2-ethylhexyl) sulfosuccinate, AOT). With the aid of two-photon fluorescence (TPF), Rayleigh scattering and TEM, we revealed the structural evolution of the aggregated micelles/vesicles. It was found that AOT and DOX molecules rapidly aggregated and formed micelles in the solution. The residual DOX then acted as a "glue" that induced the aggregating/growing of the micelles and the transformation from aggregates to vesicles. The existence of lipid films, which was considered as the necessary intermediate state for vesicle formation, was excluded via the SHG observations, indicating that hollow shells may be directly transformed from solid aggregated micelles in the self-assembly formation of complex vesicles. The combined spectroscopic methods were also used to investigate the formation of vesicles from a commonly used lipid (i.e., 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt, DOPG) from its stacked bilayers. The swelling, curving and sealing of the DOPG bilayers for vesicle formation was monitored and clear dynamics were revealed. This work shows that the vesicle formation mechanism varies with the initial state of the surfactant/lipid molecules. It not only demonstrates the capability of the combined spectroscopic methods in investigating the aggregated systems but also provides new insight for understanding the formation of vesicles.

Probing catalytic reduction on the surface of Au nanoparticles by second-harmonic generation and two-photon luminescence.

The catalytic reduction of aromatic nitro compounds by metallic nanoparticles in the presence of sodium borohydride (NaBH4) has been widely studied as model reactions. However, the reaction mechanisms still need further investigations. For example, the origin of the induction time that has often been observed is still controversial. Here, we demonstrated that such catalytic reduction reactions on the surface of colloidal gold nanoparticles (AuNPs) may be inspected by the second-harmonic generation (SHG) and two-photon luminescence (TPL) emission from AuNPs. It was revealed that the SHG and TPL signals from AuNPs were sensitive to the substitution of citrate by active hydride species derived from the hydrolysis of NaBH4. Based on the UV-vis spectroscopy analyses and monitoring the SHG/TPL signals, the induction time in the catalytic reaction of 4-nitrothiophenol was revealed to originate from the hindered adsorption of hydride on the gold surface. This work demonstrated that SHG and TPL can provide a new approach for detecting active hydrides on the surface of metallic nanoparticles in colloids.

Combined Second Harmonic Generation and Fluorescence Analyses of the Structures and Dynamics of Molecules on Lipids Using Dual-Probes: A Review.

Revealing the structures and dynamic behaviors of molecules on lipids is crucial for understanding the mechanism behind the biophysical processes, such as the preparation and application of drug delivery vesicles. Second harmonic generation (SHG) has been developed as a powerful tool to investigate the molecules on various lipid membranes, benefiting from its natural property of interface selectivity, which comes from the principle of even order nonlinear optics. Fluorescence emission, which is in principle not interface selective but varies with the chemical environment where the chromophores locate, can reveal the dynamics of molecules on lipids. In this contribution, we review some examples, which are mainly from our recent works focusing on the application of combined spectroscopic methods, i.e., SHG and two-photon fluorescence (TPF), in studying the dynamic behaviors of several dyes or drugs on lipids and surfactants. This review demonstrates that molecules with both SHG and TPF efficiencies may be used as intrinsic dual-probes in plotting a clear physical picture of their own behaviors, as well as the dynamics of other molecules, on lipid membranes.

Unveiling the Molecular Dynamics in a Living Cell to the Subcellular Organelle Level Using Second-Harmonic Generation Spectroscopy and Microscopy.

Second-harmonic generation (SHG) microscopy has been proved to be a powerful method for investigating the structures of biomaterials. SHG spectra were also generally used to probe the adsorption and cross-membrane transport of molecules on lipid bilayers in situ and in real time. In this work, we applied SHG and two-photon fluorescence (TPF) spectra to investigate the dynamics of an amphiphilic ion with an SHG and TPF chromophore, D289 (4-(4-diethylaminostyry)-1-methyl-pyridinium iodide), on the surface of human chronic myelogenous leukemia (K562) cells and the subcellular structures inside the cells. The adsorption and cross-membrane transport of D289 into the cells and then into the organelles such as mitochondria were revealed. SHG images were also recorded and used to demonstrate their capability of probing molecular dynamics in organelles in K562 cells. This work demonstrated the first SHG investigation of the cross-membrane transport dynamics on the surface of subcellular organelles. It may also shed light on the differentiation of different types of subcellular structures in cells.

MOF Structure Engineering to Synthesize CoNC Catalyst with Richer Accessible Active Sites for Enhanced Oxygen Reduction.

Single-atom cobalt-based CoNC are promising low-cost electrocatalysts for oxygen reduction reaction (ORR). However, further increasing the single cobalt-based active sites and the ORR activity remain a major challenge. Herein, an acetate (OAc) assisted metal-organic framework (MOF) structure-engineering strategy is developed to synthesize hierarchical accordion-like MOF with higher loading amount and better spatial isolation of Co and much higher yield when compared with widely reported polyhedron MOF. After pyrolysis, the accordion-structured CoNC (CoNC (A)) is loaded with denser CoN4 active sites (Co: 2.88 wt%), approximately twice that of Co in the CoNC reported. The presence of OAc in MOF also induces the generation of big pores (5-50 nm) for improving the accessibility of active sites and mass transfer during catalytic reactions. Consequently, the CoNC (A) catalyst shows an admirable ORR activity with a E1/2 of 0.89 V (40 mV better than Pt/C) in alkaline electrolytes, outstanding durability, and absolute tolerance to methanol in both alkaline and acidic media. The CoNC-based Zn-air battery exhibits a high specific capacity (976 mAh g-1 Zn ), power density (158 mW cm-2 ), rate capability, and long-term stability. This work demonstrates a reliable approach to construct single atom doped carbon catalysts with denser accessible active sites through MOF structure engineering.

Photocatalytic redox on the surface of colloidal silver nanoparticles revealed by second harmonic generation and two-photon luminescence.

The redox of silver on the surface of Ag nanoparticles (AgNPs) has received extensive attention because of its significant impact on the biological, physical and chemical properties of AgNPs and their applications. Here we demonstrate that the surface redox reaction of AgNPs in colloids may be investigated by the second harmonic generation (SHG) and two-photon luminescence (TPL) emission from the AgNPs. It was revealed that the oxidation of silver on the surface of AgNPs was accelerated upon femtosecond laser excitation, accompanied by a decrease in the SHG and TPL emissions from the AgNPs. The photon-induced reduction of oxidized silver on AgNPs and the formation of surface defects were also revealed by the changes in the SHG and TPL emissions. Size and morphology changes have not been detected by dynamic light scattering and TEM measurements. The changes in the UV-vis extinction spectra were also very weak compared with previous reports. However, the occurrence of redox reactions on the Ag surface upon femtosecond laser irradiation has been confirmed by multiple control experiments. This work demonstrates that SHG and TPL can sensitively probe the subtle structural change on the surface of AgNPs.

Particle adsorption at the oil-water interface studied with second harmonic generation.

In this work, energetics of the adsorption of polystyrene nanoparticles at the hexadecane-water interface was studied with second harmonic generation. The adsorption of positively and negatively charged nanoparticles at the oil-water interface induced a decrease and an increase in the SHG emission from the interface, respectively. This change in the SHG emission, which is similar to that upon the adsorption of ionic surfactants at the hexadecane-water interface, which we reported previously, was then used as an indicator of particle adsorption at the interface. The adsorption free energies of the particles with a diameter of 20 nm at the hexadecane-water interface were found to be -14.7 ± 0.5 kcal mol-1, -14.4 ± 0.4 kcal mol-1 and -15.1 ± 0.3 kcal mol-1 for the amidine, carboxyl and sulfate latex beads, respectively. This result implied that the van der Waals interaction between the oil phase and the polystyrene particles is capable of driving negatively charged particles to the negatively charged hexadecane-water interface. The principle of like dissolves like played a major role in the adsorption of polystyrene particles from the aqueous phase to the oil-water interface. The origin of the SHG emission from the oil-water interface was also discussed.

Nano oxide intermediate layer assisted room temperature sintering of ink-jet printed silver nanoparticles pattern.

Sintering of metallic nanoparticles (NPs) at low temperature is highly wanted in the manufacturing of flexible electronics. And for ink-jet printing, the metallic NPs after printing usually need thermal or chemical post-treatment to remove stabilizing agents and achieve conductivity. Here, we reported a facile method to realize one-step printed sintering of silver nanoparticle (AgNP) ink at room temperature by using intermediate coated layers composed of oxide NPs and polyvinyl alcohol (PVA) mixture. We found that the detachment of the stabilizer (citrate) from the AgNPs was caused by hydroxyl groups on the surface of the oxide NPs, which enabled the coalescence and sintering of the AgNPs. With the aid of SiO2 NPs based intermediate layer, the patterns showed resistivity as low as 3.45 µΩ cm after sintering. Moreover, the mixed PVA could ensure the forming quality of patterns owing to its adsorption of ink and the high adhesiveness of PVA with substrates. So, we envision that this approach could serve as an adaptive method for sintering of AgNPs based conductive patterns on various substrates at room temperature and promote the manufacture of printed electronics.

Surface curvature-dependent adsorption and aggregation of fluorescein isothiocyanate on gold nanoparticles.

The interaction between metallic nanoparticles and fluorescent molecules and its influence on the optical properties of the particles/molecules have been intensively investigated because of their biology and sensing applications. Here, we studied the adsorption and aggregation of a commonly used dye, fluorescein isothiocyanate (FITC), on gold nanoparticles of various diameters. It was observed that the adsorption of FITC on relatively large gold nanoparticles (≥15 nm in diameter) induced quenching in the two-photon fluorescence (TPF) emission from the FITC molecules, while smaller-sized gold nanoparticles (1.6 nm) had no such effect. This difference was interpreted by the fluorescence resonance energy transfer (FRET) between the FITC molecules and the larger gold nanoparticles. At the same time, it was observed that the ratio of TPF quenching was notably higher than the ratio of the FITC molecules chemically adsorbed on the large gold particles. This unexpected observation revealed that the aggregation-induced fluorescence quenching also contributed significantly to the attenuation of the TPF emission. Time-dependent TPF attenuation during the interaction of FITC and the larger gold nanoparticles was recorded and used to confirm this interpretation. With this experimental evidence, a clear picture of the interaction of the FITC molecules on the gold surface was presented: FITC molecules chemically adsorbed on the small gold nanoparticles. However, the relatively larger surface curvature hindered the aggregation of the FITC molecules on the small gold nanoparticles. On the surface of the larger gold nanoparticles, both adsorption and aggregation occured. The influence of the surface curvature on the interfacial structure of the adsorbed molecules on nanoparticles was discussed.

Drastically modulating the structure, fluorescence, and functionality of doxorubicin in lipid membrane by interfacial density control.

In this work, we report on the observation of a drastic modulation of the fluorescence emission of an anticancer drug, doxorubicin, at the lipid interface during the variation of its molecular density at the interface. The emission efficiency of doxorubicin in the lipid membrane was modulated in the range of less than 10% to above 300% that in the aqueous solution. The corresponding changes in the structure and functionality of doxorubicin on the lipid surface were analyzed with the aid of second harmonic generation and theoretical calculation. It was observed that doxorubicin molecules aggregated on the lipid membrane at a relatively high interfacial density. However, this aggregation may not cause interfacial domain large enough to alter the permeability of the lipid bilayer. At an even higher doxorubicin density, the domain of the aggregated doxorubicin molecules induced a cross-membrane transportation.

Preparation of reduced graphite oxide loaded with cobalt(II) and nitrogen co-doped carbon polyhedrons from a metal-organic framework (type ZIF-67), and its application to electrochemical determination of metronidazole.

The integration of derivatives of granular metal-organic frameworks (MOFs) and an electrically conductive carbon substrate is an effective way to circumvent the deficiency of powdered pristine MOFs or MOF-derived carbon in practical application. The authors describe the use of graphite oxide (GO) as a substrate for in-situ assembly with the zeolitic imidazole framework ZIF-67. The GO and ZIF-67 composites were converted, via pyrolysis, into reduced graphite oxide loaded with Co/N-co-doped carbon polyhedrons (ZIF-67C@rGO). By using various amounts of GO, a series of ZIF-67C@rGO-x with different fractions of GO were synthesized and utilized as electrode modifiers for the detection of the antibiotic metronidazole (MNZ). The results revealed that the ZIF-67C@rGO-0.06 display best sensing performance. This is likely to be due to its hierarchically open pores, abundant active sites and good electrical conductivity. The sensor, best operated near a working potential around -0.6 V (vs. SCE), has a linear response in the 0.5 to 1000 µM MNZ concentration range and a 0.05 µM detection limit. The sensor was applied to the analysis of pharmaceutical samples where it showed excellent selectivity, good repeatability and satisfying recoveries. Graphical abstract Schematic representation of preparation and application of ZIF-67C@rGO-x.