Supramolecular optimization of the visual contrast for colorimetric indicator assays that release resorufin dye.
Ontology highlight
ABSTRACT:
SUBMITTER: Morsby JJ
PROVIDER: S-EPMC7429340 | biostudies-literature |
REPOSITORIES: biostudies-literature
Similar Datasets
Project description:Amplex Red is a fluorescent probe that is widely used to detect hydrogen peroxide (H2O2) in a reaction where it is oxidised to resorufin by horseradish peroxidase (HRP) as a catalyst. This assay is highly rated amongst other similar probes thanks to its superior sensitivity and stability. However, we report here that Amplex Red is readily converted to resorufin by a carboxylesterase without requiring H2O2, horseradish peroxidase or oxygen: this reaction is seen in various tissue samples such as liver and kidney as well as in cultured cells, causing a serious distortion of H2O2 measurements. The reaction can be inhibited by Phenylmethyl sulfonyl fluoride (PMSF) at concentrations which do not disturb mitochondrial function nor the ability of the Amplex Red-HRP system to detect H2O2.In vitro experiments and in silico docking simulations indicate that carboxylesterases 1 and 2 recognise Amplex Red with the same kinetics as carboxylesterase-containing mitochondria. We propose two different approaches to correct for this problem and re-evaluate the commonly performed experimental procedure for the detection of H2O2 release from isolated liver mitochondria. Our results call for a serious re-examination of previous data.
| S-EPMC4708625 | biostudies-literature
Project description:The facile coupling of azobenzene dyes to the quadruply hydrogen-bonding modules 2,7-diamido-1,8-naphthyridine (DAN) and 7-deazaguanine urea (DeUG) is described. The coupling of azobenzene dye 2 to mono-amido DAN units 4, 7, and 9 was effected by classic 4-(dimethylamino)pyridine (DMAP)-catalyzed peptide synthesis with N-(3-dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride (EDC) as activating agent, affording the respective amide products 5, 8, and 10 in 60-71% yield. The amide linkage was formed through either the aliphatic or aromatic ester group of 2, allowing both the flexibility and absorption maximum to be tuned. Azobenzene dye 1 was coupled to the DeUG unit 11 by Steglich esterification to afford the product amide 12 in 35% yield. Alternatively, azobenzene dye 16 underwent a room-temperature copper-catalyzed azide-alkyne Huisgen cycloaddition with DeUG alkyne 17 to give triazole 18 in 71% yield. Azobenzene coupled DAN modules 5, 8, and 10 are bright orange-red in color, and azobenzene coupled DeUG modules 12 and 18 are orange-yellow in color. Azobenzene coupled DAN and DeUG modules were successfully used as colorimetric indicators for specific DAN-DeUG and DAN-UPy (2-ureido-4(1H)-pyrimidone) quadruply hydrogen-bonding interactions.
| S-EPMC3326628 | biostudies-literature
Project description:Rapid and sensitive detection of pathogens is a key requirement for both environmental and clinical settings. We report here a colorimetric enzyme-nanoparticle conjugate system for detection of microbial contamination. In this approach, cationic gold nanoparticles (NPs) featuring quaternary amine headgroups are electrostatically bound to an enzyme [?-galactosidase (?-Gal)], inhibiting enzyme activity. Analyte bacteria bind to the NP, which releases the ?-Gal and restores its activity, providing an enzyme-amplified colorimetric readout of the binding event. Using this strategy, we have been able to quantify bacteria at concentrations of 1 × 10(2) bacteria/mL in solution and 1 × 10(4) bacteria/mL in a field-friendly test strip format.
| S-EPMC3120917 | biostudies-literature
Project description:In this work, a low-cost, portable device is developed to detect colorimetric assays for in-field and point-of-care (POC) analysis. The device can rapidly detect both pH values and nitrite concentrations of five different samples, simultaneously. After mixing samples with specific reagents, a high-resolution digital camera collects a picture of the sample, and a single-board computer processes the image in real time to identify the hue-saturation-value coordinates of the image. An internal light source reduces the effect of any ambient light so the device can accurately determine the corresponding pH values or nitrite concentrations. The device was purposefully designed to be low-cost, yet versatile, and the accuracy of the results have been compared to those from a conventional method. The results obtained for pH values have a mean standard deviation of 0.03 and a correlation coefficient R2 of 0.998. The detection of nitrites is between concentrations of 0.4-1.6?mg l-1, with a low detection limit of 0.2?mg l-1, and has a mean standard deviation of 0.073 and an R2 value of 0.999. The results represent great potential of the proposed portable device as an excellent analytical tool for POC colorimetric analysis and offer broad accessibility in resource-limited settings.
| S-EPMC5717667 | biostudies-literature
Project description:The localization and measurement of neuronal activity magnitude at high spatial and temporal resolution are essential for mapping and better understanding neuronal systems and mechanisms. One such example is the generation of retinotopic maps, which correlates localized retinal stimulation with the corresponding specific visual cortex responses. Here we evaluated and compared seven different methods for extracting and localizing cortical responses from voltage-sensitive dye imaging recordings, elicited by visual stimuli projected directly on the rat retina by a customized projection system. The performance of these methods was evaluated both qualitatively and quantitatively by means of two cluster separation metrics, namely, the (adjusted) Silhouette Index (SI) and the (adjusted) Davies-Bouldin Index (DBI). These metrics were validated using simulated data, which showed that Temporally Structured Component Analysis (TSCA) outperformed all other analysis methods for localizing cortical responses and generating high-resolution retinotopic maps. The analysis methods, as well as the use of cluster separation metrics proposed here, can facilitate future research aiming to localize specific activity at high resolution in the visual cortex or other brain areas.
| S-EPMC8490879 | biostudies-literature
Project description:A new class of dye-sensitized solar cells (DSSCs) using the hemicage cobalt-based mediator [Co(ttb)]2+/3+ with the highly preorganized hexadentate ligand 5,5'',5''''-((2,4,6-triethyl benzene-1,3,5-triyl)tris(ethane-2,1-diyl))tri-2,2'-bipyridine (ttb) has been fully investigated. The performances of DSSCs sensitized with organic D-?-A dyes utilizing either [Co(ttb)]2+/3+ or the conventional [Co(bpy)3 ]2+/3+ (bpy=2,2'-bipyridine) redox mediator are comparable under 1000?W?m-2 AM 1.5?G illumination. However, the hemicage complexes exhibit exceptional stability under thermal and light stress. In particular, a 120-hour continuous light illumination stability test for DSSCs using [Co(ttb)]2+/3+ resulted in a 10?% increase in the performance, whereas a 40?% decrease in performance was found for [Co(bpy)3 ]2+/3+ electrolyte-based DSSCs under the same conditions. These results demonstrate the great promise of [Co(ttb)]2+/3+ complexes as redox mediators for efficient, cost-effective, large-scale DSSC devices.
| S-EPMC5305181 | biostudies-literature
Project description:We report on the adaptation of a smartphone's rear-facing camera to function as a spectrometer that measures the spectrum of light scattered by common paper-based assay test strips. We utilize a cartridge that enables a linear series of test pads in a single strip to be swiped past the read head of the instrument while the phone's camera records video. The strip is housed in a custom-fabricated cartridge that slides through the instrument to facilitate illumination with white light from the smartphone's flash LED that is directed through an optical fiber. We demonstrate the ability to detect subtle changes in the scattered spectrum that enables quantitative analysis of single-analyte and multi-analyte strips. The demonstrated capability can be applied to broad classes of paper-based assays in which visual observation of colored strips is not sufficiently quantitative, and for which analysis of red-green-blue pixel values of a camera image are not capable of measuring complex scattered spectra.
| S-EPMC6774390 | biostudies-literature
Project description:A simple and versatile colorimetric biosensor has been developed for sensitive and specific detection of a wide range of biomolecules, such as oligonucleotides and aptamer-recognized targets. Combining the signal transducer and catalyzed hairpin assembly (CHA)-based signal amplification, the target DNA binds with the hairpin DNA to form a new nucleic acid sequence and creates a toehold in the transducer for initiating the recycle amplification reaction of CHA. The catalyzed assembly process produces a large amount of G-rich DNA. In the presence of hemin, the G-rich DNA forms G-quadruplex/hemin complex and mimic horseradish peroxidase activity, which catalyzes a colorimetric reaction. Under optimal conditions, the calibration curve of synthetic target DNA has good linearity from 50 pM to 200 nM with a detection limit of 32 pM. This strategy has been successfully applied to detect S. pneumoniae as low as 156 CFU mL(-1), and shows a good specificity against closely related streptococci and major pathogenic bacteria. In addition, the developed method enables successful visual analysis of S. pneumoniae in clinical samples by the naked eye. Importantly, this method demonstrates excellent assay versatility for sensitively detecting oligonucleotides or aptamer-recognized targets.
| S-EPMC4462091 | biostudies-literature
Project description:Paper-based analytical devices (PADs) employing colorimetric detection and smartphone images have gained wider acceptance in a variety of measurement applications. PADs are primarily meant to be used in field settings where assay and imaging conditions greatly vary, resulting in less accurate results. Recently, machine-learning (ML)-assisted models have been used in image analysis. We evaluated a combination of four ML models-logistic regression, support vector machine (SVM), random forest, and artificial neural network (ANN)-as well as three image color spaces, RGB, HSV, and LAB, for their ability to accurately predict analyte concentrations. We used images of PADs taken at varying lighting conditions, with different cameras and users for food color and enzyme inhibition assays to create training and test datasets. The prediction accuracy was higher for food color than enzyme inhibition assays in most of the ML models and color space combinations. All models better predicted coarse-level classifications than fine-grained concentration classes. ML models using the sample color along with a reference color increased the models' ability to predict the result in which the reference color may have partially factored out the variation in ambient assay and imaging conditions. The best concentration class prediction accuracy obtained for food color was 0.966 when using the ANN model and LAB color space. The accuracy for enzyme inhibition assay was 0.908 when using the SVM model and LAB color space. Appropriate models and color space combinations can be useful to analyze large numbers of samples on PADs as a powerful low-cost quick field-testing tool.
| S-EPMC8675014 | biostudies-literature
Project description:To improve the performance of dye-sensitized photoelectrochemical cell (DS-PEC) devices for splitting water, the tailoring of the photocatalytic four-photon water oxidation half-reaction represents a principle challenge of fundamental significance. In this study, a Ru-based water oxidation catalyst (WOC) covalently bound to two 2,6-diethoxy-1,4,5,8-diimide-naphthalene (NDI) dye functionalities provides comparable driving forces and channels for electron transfer. Constrained ab initio molecular dynamics simulations are performed to investigate the photocatalytic cycle of this two-channel model for photocatalytic water splitting. The introduction of a second light-harvesting dye in the Ru-based dye-WOC-dye supramolecular complex enables two separate parallel electron-transfer channels, leading to a five-step catalytic cycle with three intermediates and two doubly oxidized states. The total spin S=1 is conserved during the catalytic process and the system with opposite spin on the oxidized NDI proceeds from the Ru=O intermediate to the final Ru-O2 intermediate with a triplet molecular 3 O2 ligand that is eventually released into the environment. The in-depth insight into the proposed photocatalytic cycle of the two-channel model provides a strategy for the development of novel high-efficiency supramolecular complexes for DS-PEC devices with buildup and conservation of spin multiplicity along the reaction coordinate as a design principle.
| S-EPMC8453919 | biostudies-literature