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Colorimetric sensor array for soft drink analysis.

Fourteen commercial soft drinks have been analyzed using colorimetric sensor arrays made from a set of 25 chemically responsive dyes printed on a hydrophobic membrane. Digital imaging of the dye array before and after immersion provides a color change profile as a unique fingerprint for each specific analyte. The digital data library generated was analyzed with statistical and chemometric methods, including principal component analysis (PCA) and hierarchical clustering analysis (HCA). Facile identification of all of the soft drinks was readily achieved using comparison of the color change profiles or a PCA score plot. Using a HCA dendrogram, the misclassification rate was <2%, and even very similar sodas were easily differentiated. In addition, the monitoring of soft drinks as they degas or upon dilution also proved to be possible. This work demonstrates the potential of our colorimetric sensor array technology for quality assurance/control applications of sodas and perhaps other beverages as well.

Carbonated Beverages↗

Colorimetric sensor arrays for the analysis of beers: a feasibility study.

Eighteen commercial beers have been analyzed in both liquid and gas phases using colorimetric sensor arrays made from selected chemically responsive dyes printed on a hydrophobic membrane. Digital imaging of the dye array before and after exposure to the complex analytes in either the liquid phase or the head-gas provides a color change profile as a unique fingerprint for the specific analyte. The digital data libraries generated were analyzed using statistical and chemometric methods, including principal component analysis (PCA) and hierarchical clustering analysis (HCA). In either liquid- or gas-phase experiments, facile identification of specific beers was achieved using comparison of the color change profiles; using HCA statistical analysis the error rate of identification was <3%. Differentiation between even very similar beers proved to be straightforward. In addition, differentiation of pristine beer from the effects of watering or decarbonation proved to be possible. These results suggest that colorimetric sensor arrays may prove to be useful for quality assurance/quality control applications of beers and perhaps other beverages.

Beer↗

Colorimetric sensor arrays for volatile organic compounds.

The development of a low-cost, sensitive colorimetric sensor array for the detection and identification of volatile organic compounds (VOCs) is reported. Using an array composed of chemoresponsive dyes, enormous discriminatory power is possible in a simple device that can be imaged easily with an ordinary flatbed scanner. Excellent differentiation of closely related organic compounds can be achieved, and a library of 100 VOCs is presented. The array discriminates among VOCs by probing a wide range of intermolecular interactions, including Lewis acid/base, Brønsted acid/base, metal ion coordination, hydrogen bonding, and dipolar interactions. Importantly, by proper choice of dyes and substrate, the array is essentially nonresponsive to changes in humidity.

Biosensing Techniques↗

A colorimetric sensor array for organics in water.

Molecular recognition of organic compounds in aqueous solutions is inherently challenging due to the potential interference from the very high concentration of water. Here we present a simple colorimetric sensor array that probes a wide range of chemical properties. By printing hydrophobic dyes on a hydrophobic membrane, sensor arrays are easily prepared that provide substantial chemical selectivity for the identification and quantification of various organics (both single compounds and complex mixtures) dissolved in water. It is possible to differentiate easily even among closely related organic compounds. Upon immersion in aqueous solutions, digital imaging of the dye array before and after exposure to an analyte provide a color change profile that is a fingerprint for the organic components of the solution. Facile identification of a wide variety of aqueous organic solutions is possible over a concentration range of 0.1 M to 1 muM. Complex mixtures present no inherent difficulty; for example, a series of commercial soft drinks were easily distinguished using the colorimetric sensor array approach.

Colorimetry↗

Machine learning-assisted Mn-N-C nanozyme colorimetric sensor array for trace-level detection of biogenic amines in meat.

Accurate detection of biogenic amines (BAs) in meat remains challenging due to their high structural similarity and co-occurrence. Herein, an Mn-N-C nanozyme was synthesized via a metal-organic framework confined pyrolysis strategy, possessing excellent oxidase (OXD)- and peroxidase (POD)-like activities. The dual enzyme-like activity showed Km values of 0.1584&#xa0;mM (OXD) and 0.1498&#xa0;mM (POD), respectively, in detection system. Leveraging these properties, a colorimetric sensor array was constructed, enabling the detection of four representative BAs within a concentration range of 2-10&#xa0;ppm with 100% classification accuracy. In addition, a concentration independent recognition model based on an artificial neural network was developed to address signal nonlinearity interference in meat. The integrated system achieved accurate trace-level identification of BAs in perishable fish, pork, and chicken, demonstrating its applicability for early-stage BAs monitoring and quality deterioration warning during storage and transportation.

Biogenic Amines↗

An optoelectronic nose: "seeing" smells by means of colorimetric sensor arrays.

A new approach to general sensors for odors and volatile organic compounds (VOCs) using thin films of chemically responsive dyes as a colorimetric sensor array is described. This optoelectronic "nose," by using an array of multiple dyes whose colors change based on the full range of intermolecular interactions, provides enormous discriminatory power among odorants in a simple device that can be easily digitally imaged. High sensitivities (ppb) have been demonstrated for the detection of biologically important analytes such as amines, carboxylic acids, and thiols. By the proper choice of dyes and substrate, the array can be made essentially nonresponsive to changes in humidity.

Biosensing Techniques↗

A colorimetric sensor array for odour visualization.

Array-based vapour-sensing devices are used to detect and differentiate between chemically diverse analytes. These systems--based on cross-responsive sensor elements--aim to mimic the mammalian olfactory system by producing composite responses unique to each odorant. Previous work has concentrated on a variety of non-specific chemical interactions to detect non-coordinating organic vapours. But the most odiferous, toxic compounds often bind readily to metal ions. Here we report a simple optical chemical sensing method that utilizes the colour change induced in an array of metalloporphyrin dyes upon ligand binding while minimizing the need for extensive signal transduction hardware. The chemoselective response of a library of immobilized vapour-sensing metalloporphyrin dyes permits the visual identification of a wide range of ligating (alcohols, amines, ethers, phosphines, phosphites, thioethers and thiols) and even weakly ligating (arenes, halocarbons and ketones) vapours. Water vapour does not affect the performance of the device, which shows a good linear response to single analytes, and interpretable responses to analyte mixtures. Unique colour fingerprints can be obtained at analyte concentrations below 2 parts per million, and responses to below 100 parts per billion have been observed. We expect that this type of sensing array will be of practical importance for general-purpose vapour dosimeters and analyte-specific detectors (for insecticides, drugs or neurotoxins, for example).

Biosensing Techniques↗

Colorimetric molecularly imprinted polymer sensor array using dye displacement.

A colorimetric sensor array composed of seven molecularly imprinted polymers was shown to accurately identify seven different aromatic amines. The response patterns were systematically classified using linear discriminant analysis with 94% classification accuracy. Analyses of the response patterns of the analytes to the imprinted polymer array suggest that the different selectivity patterns, although subtle, appear to arise from the imprinting process. The molecular imprinting process enabled the rapid preparation of the polymers in the array from ethylene glycol dimethacrylate and methacrylic acid (80:20) in the presence of six different template molecules plus a blank nonimprinted polymer. The response of the imprinted polymer array was coupled to a colorimetric response, using a dye displacement strategy. A benzofurazan dye was selected and shown to give an accurate measure of the binding properties of the imprinted polymer array to all seven analytes. The colorimetric response also enabled the inclusion of analytes that are not spectroscopically active and were not among the original analytes that were used as template molecules. This broadens the potential utility of the imprinted polymer sensor array strategy to a wider range of analytes and applications.

Benzoxazoles↗

A microbead array chemical sensor using capillary-based sample introduction: toward the development of an "electronic tongue".

The development of a micromachined fluidic structure for the introduction of liquid samples into a chip-based sensor array composed of individually addressable polymeric microbeads is presented. The micromachined structure consists of micromachined storage cavities combined with a covering glass layer that confines the microbeads and fluidic channels. In our sensor array transduction occurs via optical (colorimetric and fluorescence) changes to receptors and indicator molecules that are covalently attached to termination sites on the polymeric microbeads. Spectral data are acquired for each of the individual microbeads using a charged-coupled device (CCD) allowing for the near-real-time analysis of liquid sample. Hence the micromachined fluidic structure must allow for both optical access to the microbeads and fluid flow through the micromachined cavities that serve as the microreactors/analysis chambers. One of the key parts of the structure is a passive fluid introduction system driven only by capillary force. This simple means of fluid introduction realizes a compact device. The capillary flow on the inlet channel has been studied, and the responses of the microbeads (alizarin complexone) to a liquid sample have been characterized. The test results show that this system is useful in a micro-total-analysis-system (mu-TAS) and biomedical applications.

Biomimetics↗

Differential receptors create patterns diagnostic for ATP and GTP.

Herein we report the combination of a library of resin-bound sensors along with a multicomponent sensor array. This novel combinatorial array sensor system shows selectivity for nucleotide phosphates in solution. The design of the anchored receptor includes a 1,3,5-trisubstituted-2,4,6-triethylbenzene scaffold coupled with peptide libraries. Each chemosensor is placed into a micromachined cavity within a silicon wafer, and the optical changes observed by a charged-coupled device result in near-real-time digital analysis of solutions. A colorimetric displacement assay was performed, and time-dependent imaging studies of the selected sensing ensembles result in a differential responses upon addition of adenosine 5'-triphosphate (ATP), adenosine 5'-monophosphate (AMP), or guanosine 5'-triphosphate (GTP). An advantage to this approach is that it creates an array of sensors that gives a fingerprint response for each analyte. Principal component analysis indicates that the library of chemosensors can differentiate between ATP, GTP, and AMP. On the basis of factor loading values, individual sensors from the library were sequenced to elucidate their chemical composition.

Adenosine Triphosphate↗

Polydiacetylene (PDA)-based colorimetric detection of biotin-streptavidin interactions.

The natural binding of streptavidin (STA) for a small molecule, biotin, has made it a useful tool in specific targeting application, due to their most specific noncovalent biological interactions in nature. Here, we describe a polydiacetylene (PDA)-based colorimetric biosensor to detect the STA-biotin interactions. We synthesized two kinds of biotin-labeled 10,12-pentacosadiynoic acid (PCDA) monomers having 2(ethylene oxide) or 3(ethylene oxide) spacer and prepared biotin-modified PDA liposomes using them. Upon the addition of STA, the modified PDA liposomes showed a color change from blue to red in about 1h followed by their aggregation and precipitation. The aggregation was found to be due to the cross-linking between the PDA liposomes caused by the STA having four biotin binding sites, which was confirmed by their TEM analysis. Since the STA-biotin interactions are widely adopted for various biological detection systems; the biotin-modified PDA sensor developed in this study has enormous potentials for the development of various colorimetric biosensors.

Acetylene↗

Dynamic monitoring of cytotoxicity on microelectronic sensors.

A real-time cell electronic sensing (RT-CES) system was used for label-free, dynamic measurement of cell responses to cytotoxicants. Cells were grown onto the surfaces of microelectronic sensors, which are comprised of circle-on-line electrode arrays and are integrated into the bottom surfaces of the microtiter plate. Changes in cell status such as cell number, viability, morphology, and adherence were monitored and quantified by detecting sensor electrical impedance. For cell quantification and viability measurement, the data generated on the RT-CES system correlated well with those from the colorimetric (MTT) assay. For cytotoxicity assessment, cells growing on microelectronic sensors were treated with different cytotoxicants, such as arsenic, mercury, and sodium dichromate. The dynamic responses of the cells to the toxicants were continuously monitored by the RT-CES system. On the basis of the IC50 values, the RT-CES system displays an equal sensitivity to the neutral red uptake assay at specific time points. Furthermore, because the RT-CES system provides real-time information regarding the state of cell morphology and adhesion in addition to cell number, we were able to discern a previously unreported effect of arsenic on NIH 3T3 cells prior to cell death. Also, using the RT-CES system, we were able to monitor cytotoxicity effects that occur within a minute of compound addition. Taken together, the RT-CES system allows for real-time, continuous monitoring and quantitative recording of the whole assay process and provides new insight into the cell-toxicant interaction.

Animals↗

A colorimetric gold nanoparticle sensor to interrogate biomolecular interactions in real time on a surface.

This paper presents a new label-free optical method to study biomolecular interactions in real time at the surface of an optically transparent substrate. The method relies on the change in the absorbance spectrum of a self-assembled monolayer of colloidal gold on glass, as a function of biomolecular binding to the surface of the immobilized colloids. Using this approach, we demonstrate proof of principle of a label-free optical biosensor to quantify biomolecular interactions in real time on a surface in a commercially available UV-visible spectrophotometer and of a colorimetric end-point assay using an optical scanner. The spectrophotometric sensor shows concentration-dependent binding and a detection limit of 16 nM for streptavidin. The sensor is easy to fabricate, is reproducible in its performance, has minimal technological requirements, namely, the availability of an UV-visible spectrophotometer or an optical scanner, and will enable high-throughput screening of biomolecular interactions in real time in an array-based format.

Biosensing Techniques↗

Development of multianalyte sensor arrays composed of chemically derivatized polymeric microspheres localized in micromachined cavities.

The development of a chip-based sensor array composed of individually addressable polystyrene-poly(ethylene glycol) and agarose microspheres has been demonstrated. The microspheres are selectively arranged in micromachined cavities localized on silicon wafers. These cavities are created with an anisotropic etch and serve as miniaturized reaction vessels and analysis chambers. A single drop of fluid provides sufficient analysis media to complete approximately 100 assays in these microetch pits. The cavities possess pyramidal pit shapes with trans-wafer openings that allows for both fluid flow through the microreactors/analysis chambers and optical access to the chemically sensitive microspheres. Identification and quantitation of analytes occurs via colorimetric and fluorescence changes to receptor and indicator molecules that are covalently attached to termination sites on the polymeric microspheres. Spectral data are extracted from the array efficiently using a charge-coupled device allowing for the near-real-time digital analysis of complex fluids. The power and utility of this new microbead array detection methodology is demonstrated here for the analysis of complex fluids containing a variety of important classes of analytes including acids, bases, metal cations, metabolic cofactors, and antibody reagents.

Chemistry Techniques, Analytical↗

Towards a fast-responding, label-free electrochemical DNA biosensor.

DNA sensors and sensor arrays (biochips) have become an important tool in molecular biology and biotechnology in recent years. For low-throughput, easy-to-use devices it is desirable that they be of low cost, reagentless, and label-free. Displacement sensors with electrochemical detection offer these advantages, and therefore the development of such a detection principle is show in this work. An HRP-labeled oligonucleotide was sub-optimally pre-hybridized with a capture probe and was displaced upon introduction of the fully complementary probe target, producing a decrease in signal that was proportional to the sample concentration. This detection scheme has been demonstrated colorimetrically and electrochemically, obtaining a total signal displacement of 55% only 5 min after introduction of the sample.

Base Sequence↗

Beta-lactamase protein fragment complementation assays as in vivo and in vitro sensors of protein protein interactions.

We have previously described a strategy for detecting protein protein interactions based on protein interaction assisted folding of rationally designed fragments of enzymes. We call this strategy the protein fragment complementation assay (PCA). Here we describe PCAs based on the enzyme TEM-1 beta-lactamase (EC: 3.5.2.6), which include simple colorimetric in vitro assays using the cephalosporin nitrocefin and assays in intact cells using the fluorescent substrate CCF2/AM (ref. 6). Constitutive protein protein interactions of the GCN4 leucine zippers and of apoptotic proteins Bcl2 and Bad, and the homodimerization of Smad3, were tested in an in vitro assay using cell lysates. With the same in vitro assay, we also demonstrate interactions of protein kinase PKB with substrate Bad. The in vitro assay is facile and amenable to high-throughput modes of screening with signal-to-background ratios in the range of 10:1 to 250:1, which is superior to other PCAs developed to date. Furthermore, we show that the in vitro assay can be used for quantitative analysis of a small molecule induced protein interaction, the rapamycin-induced interaction of FKBP and yeast FRB (the FKBP-rapamycin binding domain of TOR (target of rapamycin)). The assay reproduces the known dissociation constant and number of sites for this interaction. The combination of in vitro colorimetric and in vivo fluorescence assays of beta-lactamase in mammalian cells suggests a wide variety of sensitive and high-throughput large-scale applications, including in vitro protein array analysis of protein protein or enzyme protein interactions and in vivo applications such as clonal selection for cells expressing interacting protein partners.

3-Phosphoinositide-Dependent Protein Kinases↗