Electrochemically controlled rectification in symmetric single-molecule junctions.

Single-molecule electrochemical science has advanced over the past decades and now extends well beyond molecular imaging, to molecular electronics functions such as rectification and amplification. Rectification is conceptually the simplest but has involved mostly challenging chemical synthesis of asymmetric molecular structures or asymmetric materials and geometry of the two enclosing electrodes. Here we propose an experimental and theoretical strategy for building and tuning in situ (in operando) rectification in two symmetric molecular structures in electrochemical environment. The molecules were designed to conduct electronically via either their lowest unoccupied molecular orbital (LUMO; electron transfer) or highest occupied molecular orbital (HOMO; "hole transfer"). We used a bipotentiostat to control separately the electrochemical potential of the tip and substrate electrodes of an electrochemical scanning tunneling microscope (EC-STM), which leads to independent energy alignment of the STM tip, the molecule, and the STM substrate. By creating an asymmetric energy alignment, we observed single-molecule rectification of each molecule within a voltage range of ±0.5 V. By varying both the dominating charge transporting LUMO or HOMO energy and the electrolyte concentration, we achieved tuning of the polarity as well as the amplitude of the rectification. We have extended an earlier proposed theory that predicts electrolyte-controlled rectification to rationalize all the observed in situ rectification features and found excellent agreement between theory and experiments. Our study thus offers a way toward building controllable single-molecule rectifying devices without involving asymmetric molecular structures.

Single-molecule calorimeter and free energy landscape.

The precise measurement of thermodynamic and kinetic properties for biomolecules provides the detailed information for a multitude of applications in biochemistry, biosensing, and health care. However, sensitivity in characterizing the thermodynamic binding affinity down to a single molecule, such as the Gibbs free energy ([Formula: see text]), enthalpy ([Formula: see text]), and entropy ([Formula: see text]), has not materialized. Here, we develop a nanoparticle-based technique to probe the energetic contributions of single-molecule binding events, which introduces a focused laser of optical tweezer to an optical path of plasmonic imaging to accumulate and monitor the transient local heating. This single-molecule calorimeter uncovers the complex nature of molecular interactions and binding characterizations, which can be employed to identify the thermodynamic equilibrium state and determine the energetic components and complete thermodynamic profile of the free energy landscape. This sensing platform promises a breakthrough in measuring thermal effect at the single-molecule level and provides a thorough description of biomolecular specific interactions.

Plasmonic scattering imaging of single proteins and binding kinetics.

Measuring the binding kinetics of single proteins represents one of the most important and challenging tasks in protein analysis. Here we show that this is possible using a surface plasmon resonance (SPR) scattering technique. SPR is a popular label-free detection technology because of its extraordinary sensitivity, but it has never been used for imaging single proteins. We overcome this limitation by imaging scattering of surface plasmonic waves by proteins. This allows us to image single proteins, measure their sizes and identify them based on their specific binding to antibodies. We further show that it is possible to quantify protein binding kinetics by counting the binding of individual molecules, providing a digital method to measure binding kinetics and analyze heterogeneity of protein behavior. We anticipate that this imaging method will become an important tool for single protein analysis, especially for low volume samples, such as single cells.

Phase imaging of transition from classical to quantum plasmonic couplings between a metal nanoparticle and a metal surface.

When a metal nanoparticle is brought near to a metal surface within electron tunneling distance (∼1 nm), classical electromagnetic coupling between the nanoparticle and the metal is expected to transition to quantum coupling. We show that this transition can be observed as a drastic phase change in the surface plasmon resonance (SPR) images of the gold nanoparticles. We study the transition by controlling the distance between the nanoparticles and electrode surface, modeling the impact of the transition on the SPR image in terms of a phase shift and demonstrating detection of microRNA based on the transition from classical to quantum coupling. The work shows that the quantum coupling can be directly visualized in SPR, and the extremely sensitive dependence of the transition on distance leads to a biosensing principle with SPR.

Label-Free Quantification of Molecular Interaction in Live Red Blood Cells by Tracking Nanometer Scale Membrane Fluctuations.

Molecular interactions in live cells play an important role in both cellular functions and drug discovery. Current methods for measuring binding kinetics involve extracting the membrane protein and labeling, while the in situ quantification of molecular interaction with surface plasmon resonance (SPR) imaging mainly worked with fixed cells due to the micro-motion related noises of live cells. Here, an optical imaging method is presented to measure the molecular interaction with live red blood cells by tracking the nanometer membrane fluctuations. The membrane fluctuation dynamics are measured by tracking the membrane displacement during glycoprotein interaction. The data are analyzed with a thermodynamic model to determine the elastic properties of the cell observing reduced membrane fluctuations under fixatives, indicating cell fixations affect membrane mechanical properties. The binding kinetics of glycoprotein to several lectins are obtained by tracking the membrane fluctuation amplitude changes on single live cells. The binding kinetics and strength of different lectins are quite different, indicating the glycoproteins expression heterogeneity in single cells. It is anticipated that the method will contribute to the understanding of mechanisms of cell interaction and communication, and have potential applications in the mechanical assessment of cancer or other diseases at the single-cell level, and screening of membrane protein targeting drugs.

Transition from stochastic events to deterministic ensemble average in electron transfer reactions revealed by single-molecule conductance measurement.

Electron transfer reactions can now be followed at the single-molecule level, but the connection between the microscopic and macroscopic data remains to be understood. By monitoring the conductance of a single molecule, we show that the individual electron transfer reaction events are stochastic and manifested as large conductance fluctuations. The fluctuation probability follows first-order kinetics with potential dependent rate constants described by the Butler-Volmer relation. Ensemble averaging of many individual reaction events leads to a deterministic dependence of the conductance on the external electrochemical potential that follows the Nernst equation. This study discloses a systematic transition from stochastic kinetics of individual reaction events to deterministic thermodynamics of ensemble averages and provides insights into electron transfer processes of small systems, consisting of a single molecule or a small number of molecules.

Rapid Antimicrobial Susceptibility Testing on Clinical Urine Samples by Video-Based Object Scattering Intensity Detection.

To combat the ongoing public health threat of antibiotic-resistant infections, a technology that can quickly identify infecting bacterial pathogens and concurrently perform antimicrobial susceptibility testing (AST) in point-of-care settings is needed. Here, we develop a technology for point-of-care AST with a low-magnification solution scattering imaging system and a real-time video-based object scattering intensity detection method. The low magnification (1-2×) optics provides sufficient volume for direct imaging of bacteria in urine samples, avoiding the time-consuming process of culture-based bacterial isolation and enrichment. Scattering intensity from moving bacteria and particles in the sample is obtained by subtracting both spatial and temporal background from a short video. The time profile of scattering intensity is correlated with the bacterial growth rate and bacterial response to antibiotic exposure. Compared to the image-based bacterial tracking and counting method we previously developed, this simple image processing algorithm accommodates a wider range of bacterial concentrations, simplifies sample preparation, and greatly reduces the computational cost of signal processing. Furthermore, development of this simplified processing algorithm eases implementation of multiplexed detection and allows real-time signal readout, which are essential for point-of-care AST applications. To establish the method, 130 clinical urine samples were tested, and the results demonstrated an accuracy of ∼92% within 60-90 min for UTI diagnosis. Rapid AST of 55 positive clinical samples revealed 98% categorical agreement with both the clinical culture results and the on-site parallel AST validation results. This technology provides opportunities for prompt infection diagnosis and accurate antibiotic prescriptions in point-of-care settings.

Author Correction: Gate controlling of quantum interference and direct observation of anti-resonances in single molecule charge transport.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Interferometric plasmonic imaging and detection of single exosomes.

Exosomes play an important role in numerous cellular processes. Fundamental study and practical use of exosomes are significantly constrained by the lack of analytical tools capable of physical and biochemical characterization. In this paper, we present an optical approach capable of imaging single exosomes in a label-free manner, using interferometric plasmonic microscopy. We demonstrate monitoring of the real-time adsorption of exosomes onto a chemically modified Au surface, calculating the image intensity, and determining the size distribution. The sizing capability enables us to quantitatively measure the membrane fusion activity between exosomes and liposomes. We also report the recording of the dynamic interaction between exosomes and antibodies at the single-exosome level, and the tracking of hit-stay-run behavior of exosomes on an antibody-coated surface. We anticipate that the proposed method will contribute to clinical exosome analysis and to the exploration of fundamental issues such as the exosome-antibody binding kinetics.

Detection of Molecules and Charges with a Bright Field Optical Microscope.

Charge is a fundamental property of a molecule, and precisely measuring it enables detection of the molecule and helps understand various chemical processes involving charge. Here we show a method to measure the charge of a single nanoparticle and binding of charged molecules to the nanoparticle using a conventional bright field optical microscope. The nanoparticle is tethered to an indium tin oxide surface with a polymer and driven into oscillation with an alternating electric field, which produces scattered light captured by a camera. The weak scattered light is separated from the intense bright field background using a Fourier transform filter, and the image contrast change provides the effective charge of the nanoparticle with precision of a few electron charges or less. This method allows us to detect DNA binding to the nanoparticles, demonstrating a simple method to detect and study molecules with a conventional optical microscope.

A Paper Based Milli-Cantilever Sensor for Detecting Hydrocarbon Gases via Smartphone Camera.

Hydrocarbon gases, especially toxic ones like benzene and xylene, pose threats to human health and the environment. But existing detection techniques, like bulky GC-MS or portable PID, cannot fulfill people's requirement of affordable and reliable hydrocarbons monitoring for the purpose of personal exposure assessment. Here, a simple, low cost, and light hydrocarbon gases sensor using a smartphone camera as a readout was developed based on the paper based milli-cantilever bending induced by polymer swelling. Its sensing cantilever was composed of three layers: functional layer of polyethylene film, adhesive layer of double-side tape, and a substrate of weighing paper. And the dimensions of the milli-fabricated sensing cantilever are 8 mm long, 0.5 mm wide, and 50 µm thick. The sensor response was the displacement of milli-cantilever free end. As proof of concept, its performance to typical hydrocarbons of xylene, hexane, and BTEX was carefully examined. For all of them, the sensor showed good performance of linear response to hydrocarbon concentrations, wide detection range, low detection, and fast response. Taking xylene for example, the sensor showed wide detection range of 15-140 ppm, low detection limit of 15 ppm, and fast response of 30 s. The sensor cross-sensitivity to other hydrocarbons was consistent with polymer swelling theory that the more carbons the hydrocarbon has, the higher the sensor sensitivity. Taking advantage of the rough materials chosen and simple fabrication procedure, the developed sensors also had high stability with time, low cost, and good uniformity. The developed sensor is affordable both physically and financially, has good performance, could meet hydrocarbons monitoring requirements for occupational safety or air pollution in petroleum industry, and would benefit people's health.

A Microdroplet-Based Colorimetric Sensing Platform on a CMOS Imager Chip.

Interest in mobile chemical sensors is on the rise, but significant challenges have restricted widespread adoption into commercial devices. To be useful these sensors need to have a predictable response, easy calibration, and be integrable with existing technology, preferably fitting on a single chip. With respect to integration, the CMOS imager makes an attractive template for an optoelectronic sensing platform. Demand for smartphones with cameras has driven down the price and size of CMOS imagers over the past decade. The low cost and accessibility of these powerful tools motivated us to print chemical sensing elements directly on the surface of the photodiode array. These printed colorimetric microdroplets are composed of a nonvolatile solvent so they remain in a uniform and homogeneous solution phase, an ideal medium for chemical interactions and optical measurements. By imaging microdroplets on the CMOS imager surface we eliminated the need for lenses, dramatically scaling down the size of the sensing platform to a single chip. We believe the technique is generalizable to many colorimetric formulations, and as an example we detected gaseous ammonia with Cu(II). Limits of detection as low as 27 ppb and sensor-to-sensor variation of less than 10% across multiple printed arrays demonstrated the high sensitivity and repeatability of this approach. Sensors generated this way could share a single calibration, greatly reducing the complexity of incorporating chemical sensors into mobile devices. Additional testing showed the sensor can be reused and has good selectivity; sensitivity and dynamic range can be tuned by controlling droplet size.

Integrating Electrochemical and Colorimetric Sensors with a Webcam Readout for Multiple Gas Detection.

Multisensor detectors have merits of low cost, compact size, and capability of supplying accurate and reliable information otherwise hard to obtain by any single sensors. They are therefore highly desired in various applications. Despite the advantages and needs, they face great challenges in technique especially when integrating sensors with different sensing principles. To bridge the gap between the demand and technique, we here demonstrated an integration of electrochemical and colorimetric sensors with a webcam readout for multiple gas detection. Designed with two parallel gas channels but independent sensor cells, the dual-sensor detector could simultaneously detect each gas from their gas mixture by analysis of the group photo of the two sensors. Using Ag electro-dissolution as reporter, the bipolar electrochemical sensor achieved quantitative analysis for the first time thanks to application of pulse voltage. The sacrificed Ag layer used in the bipolar electrochemical (EC) sensor was recycled from CD, which further decreased the sensor cost and supplied a new way of CD recycling. The EC O2 sensor response, edge displacement of Ag layer due to electrochemical dissolution, has a linear relationship with O2 concentration ranging from 0 to 30% and has good selectivity to common oxidative gases. The colorimetric NO2 sensor linearly responded to NO2 concentrations ranging from 0 to 230 ppb with low detection limit of 10 ppb, good selectivity, and humidity tolerance. This integration method could be extended to integrating other gas sensors.

Direct Antimicrobial Susceptibility Testing on Clinical Urine Samples by Optical Tracking of Single Cell Division Events.

With the increasing prevalence of antibiotic resistance, the need to develop antimicrobial susceptibility testing (AST) technologies is urgent. The current challenge has been to perform the antibiotic susceptibility testing in short time, directly with clinical samples, and with antibiotics over a broad dynamic range of clinically relevant concentrations. Here, a technology for point-of-care diagnosis of antimicrobial-resistant bacteria in urinary tract infections, by imaging the clinical urine samples directly with an innovative large volume solution scattering imaging (LVSi) system and analyzing the image sequences with a single-cell division tracking method is developed. The high sensitivity of single-cell division tracking associated with large volume imaging enables rapid antibiotic susceptibility testing directly on the clinical urine samples. The results demonstrate direct detection of bacterial infections in 60 clinical urine samples with a 60 min LVSi video, and digital AST of 30 positive clinical samples with 100% categorical agreement with both the clinical culture results and the on-site agar plating validation results. This technology provides opportunities for precise antibiotic prescription and proper treatment of the patient within a single clinic visit.

Gate controlling of quantum interference and direct observation of anti-resonances in single molecule charge transport.

Quantum interference can profoundly affect charge transport in single molecules, but experiments can usually measure only the conductance at the Fermi energy. Because, in general, the most pronounced features of the quantum interference are not located at the Fermi energy, it is highly desirable to probe charge transport in a broader energy range. Here, by means of electrochemical gating, we measure the conductance and map the transmission functions of single molecules at and around the Fermi energy, and study signatures associated with constructive and destructive interference. With electrochemical gate control, we tune the quantum interference between the highest occupied molecular orbital and lowest unoccupied molecular orbital, and directly observe anti-resonance, a distinct feature of destructive interference. By tuning the molecule in and out of anti-resonance, we achieve continuous control of the conductance over two orders of magnitude with a subthreshold swing of ~17 mV dec-1, features relevant to high-speed and low-power electronics.

Intermittent photocatalytic activity of single CdS nanoparticles.

Semiconductor photocatalysis holds promising keys to address various energy and environmental challenges. Most studies to date are based on ensemble analysis, which may mask critical photocatalytic kinetics in single nanocatalysts. Here we report a study of imaging photocatalytic hydrogen production of single CdS nanoparticles with a plasmonic microscopy in an in operando manner. Surprisingly, we find that the photocatalytic reaction switches on and off stochastically despite the fact that the illumination is kept constant. The on and off states follow truncated and full-scale power-law distributions in broad time scales spanning 3-4 orders of magnitude, respectively, which can be described with a statistical model involving stochastic reactions rates at multiple active sites. This phenomenon is analogous to fluorescence photoblinking, but the underlying mechanism is different. As individual nanocatalyst represents the elementary photocatalytic platform, the discovery of the intermittent nature of the photocatalysis provides insights into the fundamental photochemistry and photophysics of semiconductor nanomaterials, which is anticipated to substantially benefit broad application fields such as clean energy, pollution treatment, and chemical synthesis.

Reliable Breathing Tracking with Wearable Mask Device.

Breathing tracking is critical for the assessment of lung functions, exercise physiologies, and energy expenditure. Conventional methods require using a face mask or mouthpiece that is connected to a stationary equipment through a tube, restricting the location, movement, or even the posture. To obtain accurate breathing physiology parameters that represent the true state of the patient during different scenarios, a wearable technology that has less intervention to patient's activities in free-living conditions is highly preferred. Here, we propose a miniaturized, reliable, and wide-dynamic ranged flow sensing technology that is immune to orientation, movement, and noise. As far as we know, this is the first work of introducing a fully integrated mask device focusing on breath tracking in free-living conditions. There are two key challenges for achieving this goal: miniaturized flow sensing and motion-induced artifacts elimination. To address these challenges, we come up with two technical innovations: 1) in hardware wise, we have designed an integrated flow sensing technique based on differential pressure Pneumotach approach and motion sensing; 2) in software wise, we have developed comprehensive algorithms based baseline tracking and orientation and motion compensation. The effectiveness of the proposed technology has been proven by the experiments. Experimental results from simulator and real breath conditions show high correlation (R2 = 0.9994 and 0.9964 respectively) and mean error within 2.5% for Minute Volume (VE), when compared to values computed from reference methods. These results show that the proposed method is accurate and reliable to track the key breath parameters in free-living conditions.

Rapid antibiotic susceptibility testing based on bacterial motion patterns with long short-term memory neural networks.

Antibiotic resistance is an increasing public health threat. To combat it, a fast method to determine the antibiotic susceptibility of infecting pathogens is required. Here we present an optical imaging-based method to track the motion of single bacterial cells and generate a model to classify active and inactive cells based on the motion patterns of the individual cells. The model includes an image-processing algorithm to segment individual bacterial cells and track the motion of the cells over time, and a deep learning algorithm (Long Short-Term Memory network) to learn and determine if a bacterial cell is active or inactive. By applying the model to human urine specimens spiked with an Escherichia coli lab strain, we show that the method can accurately perform antibiotic susceptibility testing as fast as 30 minutes for five commonly used antibiotics.

An Unobstructive Sensing Method for Indoor Air Quality Optimization and Metabolic Assessment within Vehicles.

This work investigates the use of an intelligent and unobstructive sensing technique for maintaining vehicle cabin's indoor air quality while simultaneously assessing the driver metabolic rate. CO2 accumulation patterns are of great interest because CO2 can have negative cognitive effects at higher concentrations and also since CO2 accumulation rate can potentially be used to determine a person's metabolic rate. The management of the vehicle's ventilation system was controlled by periodically alternating the air recirculation mode within the cabin, which was actuated based on the CO2 levels inside the vehicle's cabin. The CO2 accumulation periods were used to assess the driver's metabolic rate, using a model that considered the vehicle's air exchange rate. In the process of the method optimization, it was found that the vehicle's air exchange rate (λ [h-1]) depends on the vehicle speeds, following the relationship: λ = 0.060 × (speed) - 0.88 when driving faster than 17 MPH. An accuracy level of 95% was found between the new method to assess the driver's metabolic rate (1620 ± 140 kcal/day) and the reference method of indirect calorimetry (1550 ± 150 kcal/day) for a total of N = 16 metabolic assessments at various vehicle speeds. The new sensing method represents a novel approach for unobstructive assessment of driver metabolic rate while maintaining indoor air quality within the vehicle cabin.

Probing Single Molecule Binding and Free Energy Profile with Plasmonic Imaging of Nanoparticles.

Measuring binding between molecules is critical for understanding basic biochemical processes, developing molecular diagnosis, and screening drugs. Here we study molecular binding at the single molecule level by attaching nanoparticles to the molecular binding pairs. We track the thermal fluctuations of the individual nanoparticles with sub-nanometer precision using a plasmonic scattering imaging technique and show that the fluctuations are controlled by the molecular binding pairs rather than by the nanoparticles. Analysis of the thermal fluctuations provides unique information on molecular binding, including binding energy profile, effective spring constant, and switching between single and multiple molecular binding events. The method provides new insights into molecular binding and also allows one to differentiate nonspecific binding from specific binding, which has been a difficult task in biosensors.