Photoluminescence Probes in Data-Enabled Sensing.

This review summarizes the current status of development in photoluminescent probes, multidimensional photoluminescence detection, and multivariate data analysis methods. It then highlights reports featuring multivariate analysis of multidimensional measurements of photoluminescent probes published between June 2015 and June 2022, emphasizing work in the last 5 years. Important trends include the development of probe arrays, which provide fingerprint responses to the analyte(s) of interest and facilitate the analysis of complex samples; the application of neural networks and deep learning to pattern recognition and feature selection in photoluminescence images; and the application of multiway multivariate analysis to mining matrices, three-way arrays, and higher-order measurements, including hyperspectral intensity and lifetime images. These examples illustrate the increase in information extraction provided by the combination of multidimensional measurements and multivariate analysis.

Efficiency comparison of the imidazole plus RNO method for singlet oxygen detection in biorelevant solvents.

Singlet oxygen (1O2) is the focus of study in many fields, including phototoxicity, antioxidant activity, pollutant weathering, photodynamic therapy, and water disinfection. The imidazole plus RNO (Imd/RNO) method, originated by Kraljic and El Mohsni, is commonly used to monitor singlet oxygen production. In this method, 1O2 is quenched by an acceptor, imidazole (Imd), during the formation of a trans-annular peroxide intermediate that bleaches the sensor, p-nitrosodimethylaniline (RNO). Though the method has been widely used, including to monitor 1O2 production in complex environments, such as surfactants and cells, studies reporting the efficiency of the assay in complex solvents have not been reported. In this research, the Imd/RNO method in complex, biorelevant solvents, i.e., sodium dodecyl sulfate, octanol, and phosphate buffer-saturated octanol, was compared with reference solvents, i.e., phosphate buffer, ethanol, and methanol, for monitoring 1O2 produced by Rose Bengal photosensitization using time-resolved, broadband UV-Vis absorbance measurements. Rates of sensor bleaching and sensitizer photodegradation were simultaneously monitored in each solvent to investigate correlations between the disappearance rates of sensor and sensitizer. The quantum yields of 1O2 production (ϕ∆) in each solvent were calculated using a relative actinometric method. The dependence of sensor bleaching and sensitizer degradation on acceptor concentration and solvent polarity, and the results of assay controls suggest mechanistic differences underlying the reactions comprising the Imd/RNO method. These results demonstrate the need for caution and controls when using the method in complex samples including those containing cells, tissues, or nanoscale particles.

Emerging technologies for optical spectral detection of reactive oxygen species.

This review surveys recent advances in optical spectral detection of reactive oxygen species (ROS), particularly singlet oxygen, superoxide, hydroxyl radical, and hydrogen peroxide. Advances using nanoparticles and self-organizing nanostructures as well as optical detection schemes are included. Measurements using plasmonic, luminescent, photocatalytic, or self-organizing nanoparticles are highlighted. The large number of spectrophotometric and luminescent probe methods are categorized by ROS sensing mechanism, signaling mode, (de)activation mechanism, if any, and spectral chromaticity. Reports describing multicomponent ROS detection or novel nanoscale probes are discussed. Measurements using ratiometric, multichannel, or time-resolved detection and nonlinear spectral transitions are reviewed. The focus on developing probe molecules for spectral detection documented over the last 20 years has continued, with sustained emphasis on luminescence detection, but with less focus on spectrophotometric measurements. Use of nanoparticles as probes, probe carriers, and compartmentalization agents in ROS detection is increasing. On the other hand, incorporation of advanced spectral methods, such as nonlinear transition and multichannel detection, is increasing slowly in ROS analysis. This indicates there is a substantial opportunity to develop ROS measurements with use of a synergistic combination of (multi)functional nanoscale systems and advanced optical detection methods to optimize the detection limit, selectivity, and response time. Graphical abstract ᅟ.

Multivariate Analysis of Mixed Lipid Aggregate Phase Transitions Monitored Using Raman Spectroscopy.

The phase behavior of aqueous 1,2-dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC)/1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) mixtures between 8.0 ℃ and 41.0 ℃ were monitored using Raman spectroscopy. Temperature-dependent Raman matrices were assembled from series of spectra and subjected to multivariate analysis. The consensus of pseudo-rank estimation results is that seven to eight components account for the temperature-dependent changes observed in the spectra. The spectra and temperature response profiles of the mixture components were resolved by applying a variant of the non-negative matrix factorization (NMF) algorithm described by Lee and Seung (1999). The rotational ambiguity of the data matrix was reduced by augmenting the original temperature-dependent spectral matrix with its cumulative counterpart, i.e., the matrix formed by successive integration of the spectra across the temperature index (columns). Successive rounds of constrained NMF were used to isolate component spectra from a significant fluorescence background. Five major components exhibiting varying degrees of gel and liquid crystalline lipid character were resolved. Hydrogen-bonded water networks exhibiting varying degrees of organization are associated with the lipid components. Spectral parameters were computed to compare the chain conformation, packing, and hydration indicated by the resolved spectra. Based on spectral features and relative amounts of the components observed, four components reflect long chain lipid response. The fifth component could reflect the response of the short chain lipid, DHPC, but there were no definitive spectral features confirming this assignment. A minor component of uncertain assignment that exhibits a striking response to the DMPC pre-transition and chain melting transition also was recovered. While none of the spectra resolved exhibit features unequivocally attributable to a specific aggregate morphology or step in the gelation process, the results are consistent with the evolution of mixed phase bicelles (nanodisks) and small amounts of worm-like DMPC/DHPC aggregates, and perhaps DHPC micelles, at low temperature to suspensions of branched and entangled worm-like aggregates above the DMPC gel phase transition and perforated multi-lamellar aggregates at high temperature.

Phototoxic target lipid model of single polycyclic aromatic hydrocarbons.

A phototoxic target lipid model (PTLM) is developed to predict phototoxicity of individual polycyclic aromatic hydrocarbons (PAHs) measured either as median lethal concentration (LC50) or median lethal time (LT50) for a 50% toxic response. The model is able to account for the differences in the physical/chemical properties of PAHs, test species sensitivities, and variations in light source characteristics, intensity, and length of exposure. The PTLM is based on the narcotic target lipid model (NTLM) of PAHs. Both models rely on the assumption that mortality occurs when the toxicant concentration in the target lipid of the organism reaches a threshold concentration. The PTLM is applied to observed LC50s and LT50s for 20 individual PAHs, 15 test species-including arthropods, fishes, amphibians, annelids, mollusks, and algae-exposed to simulated solar and various UV light sources, for exposure times varying from less than 1 h to 100 h, a total of 333 observations. The LC50 concentrations range from less than 0.1 µg/L to greater that 104 µg/L. The model has 2 fitting parameters that are constant and apply to all PAHs and organisms. The root mean square errors of prediction for log(LC50) and log(LT50) are 0.473 and 0.382, respectively. The results indicate that the PTLM can predict the phototoxicity of single PAHs over a wide range of exposure conditions and to organisms with a wide range of sensitivities. Environ Toxicol Chem 2017;36:926-937. © 2016 SETAC.

Numerical correction of detector channel cross-talk using full-spectrum fluorescence correlation spectroscopy.

Fluorescence correlation spectroscopy (FCS) uses fluctuations in the fluorescence collected from a small illuminated volume to measure dynamic processes of fluorophores. In traditional FCS, spectral overlap produces cross-talk in dedicated detector channels, undermining the accuracy of measurements of molecular interactions. Here, the experimental realization of full-spectrum fluorescence correlation spectroscopy is described and coupled with multivariate data analysis to numerically correct detector cross-talk, isolating spectra and fluctuation traces of mixture components in spite of overlap. Application of this methodology is illustrated using the measurement of the diffusion constant of labeled polystyrene in hydroxypropyl cellulose in the presence of a persistent dye. Additionally, the results show that full-spectrum FCS with multivariate analysis can isolate and characterize signals from unanticipated sample components.

Spectral heterogeneity of PRODAN fluorescence in isotropic solvents revealed by multivariate photokinetic analysis.

This paper describes a multivariate analysis of the fluorescence emission of 6-propionyl-2-dimethylaminonaphthalene (PRODAN) in a series of isotropic solvents of differing polarity and hydrogen-bonding ability. Multivariate methods distill the essential features from spectral data matrices so that the structural details that are embedded within the data are revealed to the analyst. In the aprotic solvents investigated, the analysis reveals a pair of emission components that have emission maxima that scale with the orientational polarizability. In the alcohols, short-lived, polarity-independent blue bands tentatively attributed to neutral hydrogen-bonded solute-solvent complexes form and relax prior to emission from paired bands that have Stokes shifts that scale with the solvent hydrogen-bonding ability rather than the polarity. In water, the short-lived blue bands were not observed, but the shift in the paired bands did scale with the solvent hydrogen-bonding ability.

Photokinetic analysis of PRODAN and LAURDAN in large unilamellar vesicles from multivariate frequency-domain fluorescence.

This paper describes a multivariate photokinetic analysis of the membrane phase dependence of PRODAN and LAURDAN photokinetics in DMPC vesicles. Decay data, arranged in the form of Fourier transformed emission-decay matrices (FT-EDMs), were collected as a function of temperature around the gel phase transition temperature. Each matrix was partitioned into the emission spectra and decay profiles of the underlying emission components using methods based on principal components analysis. The analysis revealed that both probes typically emit at least three spectral components, which vary in intensity as the membrane undergoes gel to liquid-crystalline phase transitions: a locally excited species (lambda max approximately 415 nm), a charge-transfer species (lambda max approximately 435 nm), and a solvent relaxed species (lambda max approximately 490 nm). In contrast to previous reports, the most red-shifted species is not photoexcited, but evolves from the locally excited species and does not exhibit the dynamic Stokes' shifts associated with conventional solvent relaxation. The primary difference in the emission of the two probes is the prominence of the charge-transfer species in the LAURDAN emission.