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Routine Surface Plasmon Resonance (SPR) measurements, which are widely applied in affinity (bio)sensors and surface science, do not allow one to separate the obtained SPR signals into the change of the refractive index and thickness of deposited layers. For thin layers the signal is proportional to the product of the layer thickness and the difference in the refractive indices of this layer and of the aqueous media. In this study, we suggest an approach to separate these parameters. It is performed by subsequent measurements in the presence and in the absence of polyethylene glycol (PEG), an inert additive which modifies the refractive index of the liquid phase but do not penetrate into the adsorbed layer. Both parameters were successively determined for human serum albumin (HSA) as well as for anti-HSA IgG in three different settings: i) HSA adsorbed directly onto a gold surface, ii) HSA chemically immobilized on a gold surface coated with a self-assembled monolayer of 1,16 mercaptohexadecanoic acid and iii) chemically immobilized HSA with additional monomolecular layer of anti- HSA IgG antibodies bond to this protein layer. The suggested approach can be applied in most SPR devices with a flow cell for analysis of various adsorbed layers.
The electrochemical activity of 3-thienylboronic acid and its feature to form polymer films makes it a perspective receptor material for sensor applications. The affinity properties of this compound were studied here by isothermal titration calorimetry. A number of different analytes were tested, and the highest binding enthalpy was observed for sorbitol and fructose. An increase of pH in the range of 5.5–10.6 results in the rise of the binding enthalpy with an increase of the binding constant to ~8400 L/mol for sorbitol or ~3400 L/mol for fructose. The dependence of the binding constant on pH has an inflection point at pH 7.6 with a slope that is a ten-fold binding constant per one pH unit. The binding properties of 3-thienylboronic acid were evaluated to be very close to that of the phenylboronic acid, but the electrochemical activity of 3-thienylboronic acid provides a possibility of external electrical control: dependence of the affinity of 3-thienylboronic acid on its redox state defined by the presence of ferro/ferricyanide in different ratios was demonstrated. The results show that 3-thienylboronic acid can be applied in smart chemical sensors with electrochemically controllable receptor affinity.
Development of nanotechnology and corresponding industries during the last decade resulted in a new challenge for analytical science. This includes an ultrasensitive detection and characterization of nanoparticles of different origin and other nanomaterials in various media, including so complex ones as food, biological or environmental samples. The goal of this review is a systematic analysis of possible approaches and description of physical principles behind these methods. The main attention is paid to optical methods which are considered by authors to be mostly effective for the formulated task. Different approaches for detection and analysis of nanoparticles in a volume as well as of those adsorbed on a surface are discussed. While the technologies based on direct analysis of nanoparticle suspensions belong to the established approaches whose development potential has been in large extent exhausted, the novel technologies based on the surface sensing of adsorbed nanoparticles demonstrate intensive development. Therefore, the final part of the review is focused on the wide-field surface plasmon resonance microscopy. It allows one an ultrasensitive detection and characterization of individual nanoparticles of different origin in complex media and provides numerous possibilities for subsequent chemical identification of the detected particles using a hyphenation with other analytical technologies.
Graphene oxide finds applications in different fields of science, including energy conversion. Electrochemical reduction of graphene oxide (GO) significantly improves its conductivity. However, the kinetics of this process depends on the solvent, supporting electrolyte, pH, and numerous other factors. Most studies report the macroscopic views and ex-situ properties of reduced GO. To expand the knowledge about GO reduction, in this study, we used cyclic voltammetry (CV), simultaneous 2 points and 4 points resistance measurement (s24), conductive atomic force microscopy (AFM), and theoretical calculations. Using CV, we demonstrated that the choice of supporting electrolyte (KCl or LiCl) influences the potential range in which electrochemical GO reduction occurs. The activation energy of this process was estimated to be below 30 kJ mol‒1 in both electrolytes, being significantly lower than that required for thermal reduction of GO. Simultaneous in situ s24 resistance measurements suggest that GO films reach a highly conductive state at deep negative potentials, with an abrupt, irreversible switch from non-conductive to the conductive state. However, conductive AFM presents a more exact picture of this process: the reduction of GO films starts locally while the formed conductive islands grow during the reduction. This mechanism was confirmed by theoretical calculations indicating that the reduction starts on isolated oxygen-functional groups over the GO basal plane, while clustered OH groups are more difficult to reduce. The presented results can help in tailoring reduced GO for a particular electrochemical application by precisely controlling the reduction degree and percentage of the conductive area of the reduced GO films.
Modern bio-chemical sensors rely on functional interfacial architectures with well-defined structural nano-motifs over a physical transducer. Gold-coated interfaces are of particular interest for their desirable chemical (functionalization) and optical (plasmonic) properties. Here we investigate the cleaning and polishing of polycrystalline gold films in preparation of advanced surface functionalization. We focus on soft wet chemical etching to decrease the small-scale roughness commonly observed after evaporation or sputtering of gold. We show that optimized surfaces are obtained by etching in solutions of hydrochloric acid and hydrogen peroxide. We systematically quantify the films wettability, surface nano-topography, UV-VIS spectrum and the electrochemical and Surface Plasmon Resonance (SPR) changes throughout the etching process. Optimal results are obtained by etching with a HCl(37%):H2O2(30%):H2O mixture, with a volume ratio of reagents 3:3:94 during 15-20 minutes at room temperature for the main step. This reduces by a factor two the root-mean-square roughness, removes contaminants, increases hydrophilicity and modifies the gold surface by Au(Cl)x complexes. Significantly, the resulting the surface is hydrophilic enough to prevent globular proteins such as HSA to unfold upon deposition at concentrations more than ~1 mg/mL. Our protocol offers a simple, reliable and rapid method for the preparation of gold surface in view of further functionalization including the binding of receptor layers and various micro- and nanostructures required in chemical and biochemical sensing.
An analysis of early stage of electrochemical nucleation is crucial in order to understand the mechanism of this process and for electrochemical synthesis of new materials. We show here an application of the wide field surface plasmon microscopy (WF-SPRM) for this purpose. It allows us in-situ monitoring of the growth of nuclei in the range from ~20 till over 1000 nm. The system registers the moment and the place of the formation of each individual nucleus for the total number of the tracked nuclei up to ~10,000 per 1 mm2. The results demonstrate that the integral information obtained from electrochemical measurements can be extracted quantitatively from optical measurements while optical data provide also information on the size of each individual nucleus, individual nucleation time, localization and growth kinetics, and allows one to distinguish the rate limiting stage for each individual nucleus. In this work WF-SPRM was applied for investigation of electrochemical deposition of copper on gold, but this technique can be extended easily to other materials or other nucleation types.
An amperometric chemosensor for the detection of hydrogen peroxide is reported. The sensor is based on 1,4-benzoquinone immobilized on the gold electrode using self-assembled monolayer of short chain symmetrical dithiol as an anchor layer. Sensor analysis was performed by cyclic voltammetry at the potential range from −0.6 V till +0.9 V as well as in the anodic or cathodic potential ranges only. The results indicate oxidative electrochemical decomposition of hydrogen peroxide at the potential of ∼+0.4 V leading to the formation of oxygen while at cathodic potentials a reduction of the formed oxygen as well as of the hydrogen peroxide occur. A decrease in the oxidation potential of hydrogen peroxide on the gold electrode coated by self-assembled monolayer with 1,4-benzoquinone in comparison with that measured on the electrodes coated by the same self-assembled monolayer without 1,4-benzoquinone, indicates electrocatalytic effect of this moiety on oxidative decomposition of hydrogen peroxide. Analytical evaluation of the sensor performance was done in the voltammetric as well as in the chronoamperometric mode. The sensor exhibited linear response over the concentration range till 2.5 mM with a limit of detection ∼4 μM.
A concept of virtual sensor array based on an electrically controlled variation of affinity properties of the receptor layer was realized on the base of integrated electrochemical chemotransistor containing conducting polymer as the receptor layer. Electrical control of the redox-state of the polymer (polyaniline) was performed in a five-electrode configuration with four electrodes for conductivity measurements and Ag/AgCl reference electrode integrated on the same glass chip.
An ionic liquid provided an electrical connection between the reference electrode and chemosensitive material. Conductivity measurements demonstrated potential controlled electrochemical conversions of the receptor material between different redox states. The binding of trimethylamine at three different potentials corresponding to these states was studied. The results demonstrated that both kinetic- and equilibrium-binding properties of the receptor are controlled by the electrical potential, thus providing a possibility to form a virtual sensor array using only a single sensing element.
The concept was applied for monitoring fish headspace. Using three characteristics of the sensor response measured at three different redox states of the same sensor material, we obtained signals from a virtual sensor array consisting of nine chemosensitive elements. The sensor displays systematic changes of its nine signals during fish degradation. This approach can be applied also for the electrical
control of the affinity of immunoglobulins. Development of new materials with electrically controlled affinity is in progress.
Poly-3-thiopheneboronic acid was synthesized by electrochemical polymerization from 3-thienylboronic acid dissolved in the mixture of boron trifluoride diethyl etherate and acetonitrile. Cyclic voltammetry during electropolymerization shows oxidative and reductive peaks growing in each next cycle. An investigation by scanning electron microscopy displayed the polymer layer like a highly flexible film of 110 nm thick with grains of 60–120 nm in size. Strong negative solvatochromic effect was observed. Optical spectra of poly-3-thienylboronic acid at different potentials and pH were studied. Potential cycling leads to a well reversible electrochromic effect. At pH 7.4, the increase of potential leads to the decrease in the absorption band at 480 nm and to the rise in the absorption band at 810 nm with an isosbestic point at 585 nm. Spectroelectrochemical behavior of poly-3-thienylboronic acid and polythiophene was compared. Binding of sorbitol at fixed electrode potential leads to an increase in the absorbance in the shortwave band and to the decrease in the longwave band; the effect depends on the electrode potential and pH. Perspectives of application of poly-3-thienylboronic acid as new chemosensitive material are discussed.
1,3-dimercaptopropan-2-ol, a symmetrical di-thiol, has been synthesized and applied as a new type of anchor molecule to prepare a self-assembled monolayer (SAM) on the gold surface. The formed monolayers were studied by cyclic voltammetry, impedance spectroscopy, X-ray photoelectron spectroscopy, kinetic capacitance, and contact angle measurements. The SAM structure depends on the adsorption conditions. A short incubation time of the electrode at high concentration of this di-thiol leads to the predominating binding through one thiol group of the adsorbate to the gold surface, while a long incubation at low concentration leads to the predominating binding by both thiol groups. A comparative study of the desorption and replacement of SAMs indicates a strong stability increase when the SAM molecules bond gold surface by two bonds mainly. This monolayer was used to immobilize electrochemically active p-benzoquinone moiety. The surface concentration of p-benzoquinone obtained from cyclic voltammetry is 2.5 ± 0.2 × 10−10 mol·cm−2 which corresponds to the functionalization of 65 ± 5% of SAM molecules. The obtained highly stable SAM with redox-active terminal group can be applied for different tasks of chemical sensing and biosensing. As an example, an application of this system for electrocatalytical oxidation of dihydronicotinamide adenosine dinucleotide (NADH) was tested.
Despite the history of application of surface plasmon resonance (SPR) for chemo- and biosensing being over 30 years long, the development of this technique is still in progress.
This review is focused on the technological aspects of further improvement of analytical performance of SPR transducers based on Kretschmann configuration. We describe basic measurement configurations, their improvements and optimizations, and their drawbacks and limitations. An importance of referencing in SPR sensors is highlighted. The referencing approaches are classified into the following domains: (1) macroscopic spatially separated referencing, (2) self-referencing based on micro-patterning, (3) in-place referencing, (4) spatiotemporal referencing, and (5) electrochemically assisted referencing. The underlying principles of these approaches, examples of their implementation, and resulting improvements of sensor performance are described. Finally, an analysis of SPR data and an extraction of affinity properties are discussed.
The increasing energy demands of modern society require a deep understanding of the properties of energy storage materials, as well as the tuning of their performance. We show that the capacitance of graphene oxide (GO) can be precisely tuned using a simple electrochemical reduction route. In situ resistance measurements, in combination with cyclic voltammetry measurements and Raman spectroscopy, have shown that upon reduction GO is irreversibly deoxygenated, which is further accompanied by structural ordering and an increase in electrical conductivity. The capacitance is maximized when the concentration of oxygen functional groups is properly balanced with the conductivity. Any further reduction and deoxygenation leads to a gradual loss of capacitance. The observed trend is independent of the preparation route and the exact chemical and structural properties of GO. It is proposed that an improvement in the capacitive properties of any GO can be achieved by optimization of its reduction conditions.
Electrically addressed covalent immobilization of biomolecules to the defined electrodes of an electrode array is described. It is based on Michael addition of the thiol group of biomolecules to α,β-unsaturated carbonyl groups of benzoquinone. This “click” reaction was tested by immobilization of a number of thiolated compounds on the simplest array consisting of two gold electrodes coated by a self-assembled monolayer of benzoquinone-terminated hexanethiol. Electrically controlled binding of hexanethiol, ferrocenylhexanethiol, human serum albumin and thiol-terminated single-stranded DNA (ssDNA) was investigated. The binding was studied using cyclic voltammetry, X-ray photoelectron spectroscopy and surface plasmon resonance. The reaction requires the oxidized state of the benzoquinone moiety; this can be reached by applying of a moderate anodic potential to the electrode. Surface plasmon resonance measurements demonstrated that the thiol-modified ssDNA immobilized by this technique binds complementary synthetic oligonucleotides or PCR-amplified DNA fragments. The developed technology of electrical addressing of covalent immobilization can be applied for fabrication of sensor arrays.
Recently reported concept of electrical control of sensor affinity was applied for formation of a virtual sensor array based on the single sensing element. Affinity properties of this element were modulated by electrically controlled conversion of chemosensitive material between its different redox states possessing different affinity. The sensor was realized on the basis of electrochemical chemotransistor in which a low temperature ionic liquid containing chloride was used to connect the chemosensitive material to the Ag/AgCl reference electrode. The concept of virtual array was proved by its application for monitoring of fish headspace. Using three characteristics of the sensor response measured at three different redox states of the same sensor material, we have obtained signals from a virtual sensor array consisting of nine chemosensitive elements. The sensor displays systematic changes of its nine signals during fish degradation and allows us to make quantitative analysis of its freshness defined as the after the catch. The level of detection is far below the level of organoleptic detection.
The amino-substituted coumarin derivative of Tröger’s base (CTB) was electrochemically polymerized on the surface of gold electrodes. The obtained polymer films were characterized by SEM, Raman spectroscopy, cyclic voltammetry and in-situ conductometry. Spectroscopic analysis indicates that the polymerization of CTB takes place via oxidation of the aniline fragment of CTB molecule, similar to the polymerization of aniline. The similarity of the polymer backbone to that of the polyaniline is also confirmed by the influence of potential on the film’s conductivity, however the conductivity of the new material is in principle lower than that of polyaniline. SEM shows that the polymer forms a thin layer with a thickness of ~ 100 nm. Possible applications of the new material as an artificial receptor for aromatic analytes containing carboxy-group are proposed.
An approach for visualization of patterned monomolecular layers in surface plasmon microscopy (SPM) is suggested. The development of hidden image in SPM is achieved by referencing of images obtained in the presence of electrolytes with a high molar refraction of either anions or cations. A formation of diffuse layer near the charged surface areas leads to the redistribution of ions. The ratio of SPM images allows one to visualize this redistribution and to distinguish surface areas with different properties. The approach is unobtrusive and robust; it can be used with most SPR imaging instruments.
Surface plasmon resonance, being widely used in bioanalytics and biotechnology, is influenced by the electrical potential of the resonant gold layer. To evaluate the mechanism of this effect, we have studied it in solutions of various inorganic electrolytes. The magnitude of the effect decreases according to the series: KBr>KCl>KF>NaClO4. The data were treated by using different models of the interface. A quantitative description was obtained for the model, which takes into account the local dielectric function of gold being affected by the free electron charge, diffuse ionic layer near the gold/water interface, and specific adsorption of halides to the gold surface with partial charge transfer. Taking into account that most biological experiments are performed in chloride-containing solutions, detailed analysis of the model at these conditions was performed. The results indicate that the chloride adsorption is the main mechanism for the influence of potential on the surface plasmon resonance. The dependencies of surface concentration and residual charge of chloride on the applied potential were determined.
In-situ detection and characterization of nanoparticles in biological media as well as in food or other complex samples is still a big challenge for existing analytical methods. Here we describe a label-free and cost-effective analytical method for detection of nanoparticles in the concentration range 106 -1010 NPs/ml. The proposed method is based on the surface plasmon resonance microscopy (SPRM) with a large field of view (~1.3mm2 ). It is able to detect and count adsorbing nanoparticles individually, totally up to the hundreds of thousands of NPs on the sensor surface. At constant diffusion conditions the detection rate is proportional to the number concentration of NPs, this provides an approach to determine the NPs concentration. The adsorption of nanoparticle can be manipulated by the surface functionalization, pH and electrolyte concentration of suspensions. Images of detected nanoparticles can be quantified in order to characterize them individually. The image intensity grows quasi-linearly with nanoparticle size for the given material. However, the size and material of nanoparticle cannot be resolved directly from the image. For determination of chemical composition, SPRM can be assisted by electrochemical analysis. In this case, the gold sensor surface is used both as a resonant media for plasmon microscopy and as a working electrode. Under potential sweep, the adsorbed NPs can be subjected to electrochemical dissolution, which is detected optically. The potential of this conversion characterizes the material of NPs.
Detection of nano- and micro-particles is an important task for chemical analytics, food industry, biotechnology, environmental monitoring and many other fields of science and industry. For this purpose, a method based on the detection and analysis of minute signals in surface plasmon resonance images due to adsorption of single nanopartciles was developed. This new technology allows one a real-time detection of interaction of single nano- and micro-particles with sensor surface. Adsorption of each nanoparticle leads to characteristic diffraction image whose intensity depends on the size and chemical composition of the particle. The adsorption rate characterizes volume concentration of nano- and micro-particles. Large monitored surface area of sensor enables a high dynamic range of counting and to a correspondingly high dynamic range in concentration scale. Depending on the type of particles and experimental conditions, the detection limit for aqueous samples can be below 1000 particles per microliter. For application of method in complex media, nanoparticle images are discriminated from image perturbations due to matrix components. First, the characteristic SPRM images of nanoparticles (templates) are collected in aqueous suspensions or spiked real samples. Then, the detection of nanoparticles in complex media using template matching is performed. The detection of various NPs in consumer products like cosmetics, mineral water, juices, and wines was shown at sub-ppb level. The method can be applied for ultrasensitive detection and analysis of nano- and micro-particles of biological (bacteria, viruses, endosomes), biotechnological (liposomes, protein nanoparticles for drug delivery) or technical origin.
Detection of nano- and microparticles is an important task for chemical analytics, medical diagnostics, food industry, biotechnology, environmental monitoring and many other fields of science and industry. For this purpose, a method based on the detection and analysis of minute signals of surface plasmon resonance images due to adsorption of single particles was developed.
The new technology allows one a real-time detection of interaction of single nano- and microparticles of different origin with sensor surface. Adsorption of each nanoparticle leads to a characteristic diffraction image whose intensity depends on the size and chemical composition of the particle. The number of the nanoparticle - surface binding events per time and surface area characterizes volume concentration. A large monitored surface area of the sensor surface allows one to detect many hundreds events in each frame or totally up to a million particles on the sensor surface, this leads to a very high dynamic range of counting and to a correspondingly high dynamic range in the concentration scale. Depending on the type of particles and experimental conditions, the detection limit for aqueous samples can be below 1000 nanoparticles per microliter. Stable analysis of
nanoparticles in very complex environment (fruit juices, wines, cosmetic formulations) was demonstrated.
For determination of chemical composition of single nanoparticles separately, the wide field surface plasmon microscopy can be used as a tandem technique. For example, for analysis of technical nanoparticles it can be assisted by electrochemical analysis. In this case, the gold sensor surface is used both for plasmon microscopy and as a working electrode of electrochemical cell. Applying a linear potential sweep to this electrode, adsorbed nanoparticles can be subjected to an electrochemical conversion leading to the change of their refractive index; the value of electrical potential of this conversion characterizes material of the particular nanoparticle. Notably, such analysis is performed simultaneously but independently for each adsorbed particle.
The method of wide filed surface plasmon microscopy can be applied for ultrasensitive detection and analysis of nano- and microparticles of biological (bacteria, viruses, exosomes), biotechnological, (liposomes, protein nanoparticles for drug delivery) or technical (metallic, oxides, plastic, etc.) origin.
52.2016-53.2017
8.2008-17.2017
5.2014-8.2017
Microchimica Acta
(2017)
181.2014-184.2017
Sensors
(2017)
9.2009-17.2017
It has recently been shown that surface plasmon microscopy (SPM) allows single nanoparticles (NPs) on sensor surfaces to be detected and analyzed. The authors have applied this technique to study the adsorption of single metallic and plastic NPs. Binding of gold NPs (40, 60 and 100 nm in size) and of 100 nm polystyrene NPs to gold surfaces modified by differently ω-functionalized alkyl thiols was studied first. Self-assembled monolayers (SAM) with varying terminal functions including amino, carboxy, oligo(ethylene glycol), methyl, or trimethylammonium groups were deposited on gold films to form surfaces possessing different charge and hydrophobicity. The affinity of NPs to these surfaces depends strongly on the type of coating. SAMs terminated with trimethylammonium groups and carboxy group display highly different affinity and therefore were preferred when creating patterned charged surfaces. Citrate-stabilized gold NPs and sulfate-terminated polystyrene NPs were used as negatively charged NPs, while branched polyethylenimine-coated silver NPs were used as positively charged NPs. It is shown that the charged patterned areas on the gold films are capable of selectively adsorbing oppositely charged NPs that can be detected and analyzed with an ~1 ng⋅mL−1 detection limit.
Resistive gas sensors based on the composites of nanostructured carbonized polyaniline and Nafion
(2016)
Due to constant necessity to have reliable and sensitive gas sensors in many contemporary technologies, there is a permanent need for development of new sensing platforms with good sensing properties. Here, we demonstrate a novel type of resistive gas sensors based on carbonized polyaniline/Nafion composites. The sensing mechanism of such sensors is based on the sorption of gases by the composites which induce Nafion swelling and decreasing of conductivity. Chemosensitive properties can be tuned by the (i) choice of carbon materials with different conductivities, (ii) Nafion content in the composite, and (iii) thickness of the composite layer. We have shown that the sensors respond to water, acetone, ethanol, and ethanol vapors. For the last two cases, we have achieved high sensitivity, fast response, wide concentration range, and good recovery. The use of simultaneous two- and four-point techniques for these sensors provides an internal control of the sensor integrity.
A concept of virtual sensor array based on electrically controlled variation of affinity properties of the receptor layer is described. It was realized on the base of integrated electrochemical chemotransistor containing polyaniline as the receptor layer. Electrical control of the redox state of polyaniline was performed in five-electrode configuration containing four electrodes for conductivity measurements and one Ag/AgCl reference electrode. All the electrodes were integrated on the same glass chip. A room-temperature ionic liquid was used for the electrical connection between the reference electrode and chemosensitive material. Conductivity measurements demonstrated effective potential-controlled
electrochemical conversions of the receptor material between different redox states. Binding of trimethylamine at three different potentials, corresponding to the different states of the receptor material, was studied. Concentration dependencies and binding kinetics were analyzed. The results demonstrated that the kinetic as well as the equilibrium binding properties of the receptor layer can be controlled by electrical potential, thus
providing a possibility to form a virtual sensor array using only a single sensing element.
The increasing production and application of nanoparticles necessitates a highly sensitive analytical method for the quantification and identification of these potentially hazardous materials. We describe here an application of surface plasmon microscopy for the individual detection of each adsorbed nanoparticle and for visualization of its electrochemical conversion. Whereas the adsorption rate characterizes the number concentration of nanoparticles, the potential at which the adsorbed nanoparticles disappear during an anodic potential sweep characterizes the type of material. All the adsorbed nanoparticles are subjected to the potential sweep simultaneously; nevertheless, each of the up to a million adsorbed nanoparticles is identified individually by its electrochemical dissolution potential. The technique has been tested with silver and copper nanoparticles, but can be extended to many other electrochemically active nanomaterials.
Nanofibrous polyanilines (PANIs) were synthesized by several oxidative polymerization pathways that have in common the presence of excess oxidant(s) (ammonium peroxydisulfate and its mixture with hydrogen peroxide), the absence of added acid, and the absence of external template (self-assembly process). Conducting forms of the synthesized PANI nanofibers (re)doped with various acids were further used as precursors for carbonization process to obtain nanofibrous carbonaceous materials (Carb-PANIs). Morphology, molecular structure, surface properties and electrical characteristics of PANI nanofibrous precursors and their carbonized counterparts were studied by scanning electron microscopy, Raman spectroscopy, by measurements of ζ-potential and determination of isoelectric points, as well as by measurements of electrical conductivity.
Sensitive detection of engineered nanoparticles (NPs) in air and in liquid samples is an important task and still a major challenge in analytical chemistry. Recent work demonstrated that it can be performed using surface plasmon microscopy (SPM) where binding of single NPs to a surface leads to the formation of characteristic patterns in differential SPM images. However, these patterns have to be discriminated from a noisy background. Computer-assisted recognition of nanoparticles offers a solution but requires the development of respective tools for data analysis. Hereby a numerical method for automated detection and characterization of images of single adsorbing NPs in SPM image sequences is presented. The detection accuracy of the method was validated using computer generated images and manual counting. The method was applied for detecting and imaging of gold and silver NPs adsorbing from aqueous dispersions and for soot and NaCl NPs adsorbing from aerosols. The determined adsorption rate was in range 0.1–40 NPs per (s mm2) and linearly dependent on the concentration of nanoparticles. Depending on the type of NPs and signal to noise ratio, a probability of recognition of 90–95 % can be achieved.
An ultrasensitive analytical method for direct detection of single nanoparticles in complex environment is described. The method relies on the wide-field surface plasmon microscopy (SPM). The suppression of matrix effects is achieved by image analysis based on the template matching. First, characteristic SPM images of nanoparticles are collected in aqueous suspensions. Then the detection of nanoparticles in complex environment is performed using template matching.
Quantification and characterization of nanoparticles size was demonstrated at subppb level (∼ 100 pg/mL) in such complex media as wines, fruit juices, or cosmetic formulation. Visualization of the nanoparticles is performed in real time. The method does not require any sample pretreatment. If the minimally acceptable adsorption rate is defined as one nanoparticle to the whole sensor surface per few seconds, the working range of the method is ∼ 10 6 to 10 10
nanoparticles per mL.
A new approach for self-referencing in SPR biosensors is reported. The method is technologically simple and applicable for a wide range of existing SPR instrumentation with Kretschmann configuration. It is based on the micropatterning of the sensor area with sensing and referencing areas whose shape and distribution can be chosen arbitrarily and their characteristic sizes are larger than the plasmons propagation length. Provided that roughly a half of the area is used as the sensing area while its optical thickness is different from that of the referencing area, an integral measurement of the intensity of the reflected light over such a patterned surface near the summary resonance conditions exhibits self-referencing properties. An over ten-fold suppression of the effect caused by the variation of the bulk refractive index was observed.
The interaction of nanoparticles manufactured from human serum albumin (HSA) with different types of receptor surfaces was studied. Four different immobilization techniques were used to chemically link the receptors to the detector surfaces, namely: (i) a surface coated by monoclonal antibodies by a single step immobilization technique using carbodiimide, (ii) a surface coated by the same technique but using polyclonal antibodies, (iii) a surface coated by monoclonal antibodies immobilized through the sugar groups of antibodies and (iv) a surface coated by polyclonal antibodies immobilized by the same technique as in (iii).
The results showed that there was almost no difference in the affinity of HSA in the molecular form to all of these surfaces. However, the binding behaviour of nanoparticles made from the same protein was quite different as successful nanoparticle binding was only observed when the surfaces were produced using method (ii).
We report on the first application of terahertz metamaterials acting as transducers for chemical sensors based on conducting polymers. In our feasibility study aimed at sensing of gaseous hydrochloric and ammonia, a two-dimensional sensor metamaterial consisting of an array of split-ring resonators on the surface of undoped silicon wafer was prepared. The surface of the resonator was coated with a 150-μm layer of polyaniline. Binding of hydrogen chloride to polyaniline leads to distinct changes in the resonance frequency of the metamaterial. Measurements can be performed both in the reflection and transmission mode. A numerical simulation of the response revealed an increase of both the real and the imaginary components of the dielectric function of the polyaniline film. These changes are attributed to the transition from emaraldine base to emeraldine salt. The results demonstrate a new approach for formation of highly sensitive transducers for chemical sensors.
Poly(2-acrylamido-2-methyl-1-propanesulphonic acid) (PAMPSA)-doped polyaniline (PANI) layers are synthesised in the presence of sulphuric and perchloric acids. The effects of the inorganic acid as well as of the electrochemical synthetic procedure (potentiostatic and potentiodynamic deposition) and thickness of the polymer layers are studied. The focus is directed towards the pH dependence of the electrochemical redox activity and conductivity of the PAMPSA-doped PANI layers obtained under different conditions. Ascorbic acid oxidation is used as a test reaction to study the electrocatalytic behaviour of various PAMPSA-doped PANI layers in neutral solution. It is found that the type of inorganic component present in the polymerisation solution has a marked effect on the extent of doping in acidic solutions as well as on the redox electroactivity in neutral solutions. A comparison between potentiostatically and potentiodynamically synthesised layers at pH 7 shows a markedly lower conductance and lower extent of redox charge preservation in the case of potentiodynamic synthesis. The PANI electrocatalytic activity for ascorbic acid oxidation is also dependent on the polymer electrodeposition procedure, with potentiostatically synthesised layers exhibiting better electrocatalytic performance.
A clearing assay for lipolytic enzymes has been realized in 96-well microtiter plates. A thin layer containing emulsified tributyrin as turbidity-generating substrate was placed on a thicker supporting aqueous layer. Both layers were stabilized by a gel-forming agent. Enzyme addition leads to clearing of the emulsion detected with a standard microtiter plate reader as a decrease of extinction. Dependencies of the signal kinetics on the substrate and enzyme concentrations were studied. For 0.5–1 % tributyrin content the reaction rate is not substrate-limited. An initial slope of the signal kinetics is proportional to the lipase activity. A detailed characterization of the assay was performed. Lipolysis of tributyrin was confirmed by glycerol detection. Various gel-forming agents were compared and diffusion conditions in these gels were analyzed. Agar and agarose were found to be the most suitable gel-forming agents, which do not affect enzyme diffusion whereas polyacrylamide gels block lipase diffusion and therefore are not suitable for the assay. The optimized assay prepared from 1 % tributyrin emulsion in 2 % agar gel was tested with six microbial lipases and porcine pancreatic lipase. The detection limit is 20–60 ng/well which is equivalent to 30 μU/well for T. lanuginosus lipase.
Electroanalytical methods have been applied only in conducting media. An application of conducting polymers allows to overcome this limitation. If such material is in electrochemical equilibrium with dissolved redox active species, its electrical conductivity depends on the redox potential of these species. Therefore, conductometric measurements with conducting polymers can provide about the same information as classical redox electrodes. The approach was applied for redox titration. Equivalent points obtained by this titration in aqueous and organic electrolytes were identical. Then the approach was applied for determination of bromine number by redox titration in non-conducting organic phase.
Recently, developed technique for separated analysis of bulk and contact resistance was applied for the investigation of polythiophene films electropolymerized in boron trifluoride diethylether. Kinetics of polymer resistance and for the first time of the contact resistance during polymer oxidation and reduction were characterized. Influence of electrochemically controlled oxidation state on the polymer bulk and the polymer/metal contact resistance was measured in aqueous and organic environment. Variation of the electrical potential from −0.2 to 1.1 V vs. Ag/AgCl (sat) leads to an increase of the polymer conductivity for about three orders of magnitude and to a decrease of the contact resistance for about three orders of magnitude. The potential dependence of the two resistances was different, especially at high anodic potentials. In organic solution, the change of both resistances was more than six orders of magnitude. The results were compared with electrochemical and spectroelectrochemical data, a difference in the material behavior depending on the electrolyte solvent was observed. The influence of electrical potential on polymer resistance in aqueous solution was explained quantitatively by a three-state model with the values of oxidation potential +0.3 and +1.2 V.
SPR based biosensors register binding of analytes to the surface with immobilized receptors by measuring changes of the refractive index near this surface. An important task in the improvement of this measurement technology is a separation of signals, corresponding to the changes in the chemosensitive layer, from undesired contributions of bulk phase, for example, due to fluctuations of temperature, concentrations of solutes, pressure. The wavelength of the incident light influences strongly the penetration depth of the corresponding evanescent wave. This dependence was exploited here for compensation of the contribution of the bulk refractive index. It was performed using differential SPR measurements at two wavelengths with differing penetration depths. Theoretical analysis and numerical optimization of the suggested approach, named a Penetration Difference Self-Referencing SPR (PDSR-SPR), were performed. Experimental test was performed using 658 and 980 nm laser diodes. Over 20 times suppression of variations of bulk refractive index with magnitude up to 1000 μRIU was observed. Finally, PDSR-SPR approach was applied for monitoring of antibodies binding to the immobilized antigens.
A new design of conductometric chemical sensors based on conducting polymers as chemosensitive elements was suggested. The sensor includes six electrodes. Four inner electrodes coated by chemosensitive polymer are used for simultaneous two- and four-point resistance measurements thus providing information on the bulk polymer resistance and on the resistance of the polymer/electrode contacts. Two outer electrodes wired to inner electrodes by polymeric electrolyte are used for electrical control of redox state of the chemosensitive polymer. The outer electrodes are connected to potentiostat as reference and counter electrodes. It allows us to control redox state of the inner (working) electrodes. This new measurement configuration, resembling chemosensitive electrochemical transistors, provides an internal test of the sensor integrity and an electrically driven sensor regeneration. It was tested as a sensor for the detection of nitrogen dioxide. Polythiophene or polyaniline was used as receptors. Cyclic voltammograms of these polymers on the sensor surface measured in air atmosphere were very similar to that measured in aqueous electrolyte. A control of conductivity of these chemosensitive polymers by electrical potential applied vs. incorporated reference electrode was demonstrated. This effect was used for the regeneration of the chemosensitive material after exposure to nitrogen dioxide: in comparison to usual chemiresistors displaying an irreversible behavior in such test even in the time scale of hours, a completely reversible sensor regeneration within few minutes was observed.