Fluorescence Spectroscopic Methods Research Articles

Time-Resolved Fluorescence Spectroscopy of Blood, Plasma and Albumin as a Potential Diagnostic Tool for Acute Inflammation in COVID-19 Pneumonia Patients.

Fluorescence lifetime measurements of blood or plasma offer valuable insights into the microenvironment and molecular interactions of fluorophores, particularly concerning albumin. Neutrophil- and hypoxia-induced oxidative stress in COVID-19 pneumonia patients leads to hyperinflammation, various oxidative modifications of blood proteins, and potential alterations in the fluorescence lifetime of tryptophan-containing proteins, especially albumin. The objective of this study was to investigate the efficacy of time-resolved fluorescence spectroscopy of blood and plasma as a prompt diagnostic tool for the early diagnosis and severity assessment of COVID-19-associated pneumonia. This study examined a cohort of sixty COVID-19 patients with respiratory symptoms. To investigate whether oxidative stress is the underlying cause of the change in fluorescence lifetime, human serum albumin was treated with chloramine T. The time-resolved spectrometer Life Spec II (Edinburgh Instruments Ltd., Livingston, UK), equipped with a sub-nanosecond pulsed 280 nm diode, was used to measure the fluorescence lifetime of blood and plasma. The findings revealed a significant reduction in the fluorescence lifetime of blood (diluted 200 times) and plasma (diluted 20 times) at 360 nm in COVID-19 pneumonia patients compared with their respective values recorded six months post-infection and those of healthy individuals. Significant negative correlations were observed between the mean fluorescence lifetime of blood and plasma at 360 nm and several severity biomarkers and advanced oxidation protein products, while a positive correlation was found with albumin and the albumin-globulin ratio. The time-resolved fluorescence spectroscopy method demonstrates the potential to be used as a preliminary screening technique for identifying patients who are at risk of developing severe complications. Furthermore, the small amount of blood required for the measurements has the potential to enable a rapid fingerstick blood test.

3D printed carbon black-polylactic acid/copper (CB-PLA/Cu) sensor for improved sustainability in monitoring photocatalytic hydrogen peroxide evolution

As the photocatalytic generation of hydrogen peroxide continues to gather increasing interest, there is a significant need to introduce more sustainable methods to monitor the reaction process. The fabrication of low-cost sustainable electrochemical sensors offers an enticing alternative to the current standard methods. This study reports the first use of an electrochemical sensor to monitor such a process. A highly reproducible 3D printed carbon black-polylactic acid/copper sensor with an extremely low fabrication cost of €0.012 was employed to accurately monitor the photocatalytic evolution of hydrogen peroxide. The accuracy was confirmed through strong correlation with a standard fluorescence spectroscopic method. The sensor displayed limit of detection and limit of quantification values of 0.24 µM and 0.79 µM respectively, and a sensitivity of 217 µA mM−1 cm-2, while also showing strong selectivity towards hydrogen peroxide in the presence of potential interfering species. Moreover, the electrocatalytic surface was capable of ≥ 3x regeneration without significant loss in activity bringing a ≥ 75 % decrease in analysis cost and a significant reduction in potential waste material. This novel approach to monitoring the photocatalytic evolution of hydrogen peroxide presents as a more economical and sustainable alternative to current methods.

Evaluation of Groundwater Infiltration in Sewer Networks Using Fluorescence Spectroscopy

Diagnosing water infiltration is imperative to assess the integrity and operation performance of sewer networks, which is challenging and costly due to the complex nature of these networks. This study proposes a simple approach to evaluate the extent of groundwater infiltration via a fluorescence spectroscopy method, i.e., the identification and quantification of the fluorescent signature components of the dissolved organic matter sewage. A newly built sewer network in Shantou, Southern China, was selected for the case study, and a mass balance method based on water quality characteristic factors (total phosphorus and NH4+-N) was applied in parallel for comparison. The results showed that the mass balance method was substantially influenced by fluctuations in sewage and external water concentrations, rendering it unreliable due to the extensive data and calculations required. Conversely, three-dimensional excitation–emission matrix–parallel factor analysis enabled the identification of terrestrial humus compounds as the signatures of underground water sources. The estimation indicates that the groundwater proportion across the four surveyed inspection wells along the pipeline network ranged from 10.8 ± 2.5% to 9.6 ± 3.5%, conforming to the allowable groundwater infiltration limits set for municipal sewage pipelines (10–15%). This study presents a simple method for the in-depth analysis of groundwater infiltration in urban sewage networks, providing valuable insights into maintaining water quality and network integrity.

Green sustainable assessment of stability indicating first derivative synchronous fluorimetry for determination of anti-diabetic drug with its toxic impurity

Canagliflozin is prescribed with proper diet and exercise for the treatment of type 2 diabetes mellites. In this work, simple and sensitive synchronous fluorescence spectroscopic method coupled with the first derivative was proposed for the quantitative determination of canagliflozin in the presence of its oxidative degradation product. Canagliflozin and its oxidative degradation product have native fluorescence at 353 nm and 364 nm after excitation at 294 nm and 289 nm, respectively. The application of synchronous fluorescence mode gave a sharp peak but failed to resolve canagliflozin and its potential toxic degradation product. First-order derivatization of synchronous spectra resolved the overlap and allowed the determination of canagliflozin in the presence of its oxidative degradation product at 324 nm using Δλ = 20 nm. The method showed linearity with excellent correlation in the concentration range of 1–20 μg/mL for canagliflozin. The method was successfully applied for the determination of canagliflozin in pharmaceutical formulation and laboratory-prepared mixtures with mean percent recovery of 99.95 ± 1.14 and 99.81 ± 1.06, respectively. The greenness of the proposed method was assessed and approved by different tools such as analytical eco-scale, green analytical procedure index GAPI and AGREE.

Structure, interaction with biomolecules, and cytotoxicity of copper (II) complexes chelating some Schiff base ligands

Cancer remains one of the most common diseases worldwide in terms of deaths and claims many lives every day. Transition metal complexes are candidates in the development of anticancer drugs, with cisplatin being used in chemotherapy worldwide. Copper, an endogenous metal, is known for its pronounced redox potential and nucleophilicity, especially when bound to biological molecules. Cu (II) complexes were synthesized containing ethane‐1,2‐diamine as amine moiety and pentane‐2,4‐dione and/or 1‐phenylbutane‐1,3‐dione, pentane‐2,4‐dione and/or 1,1,1‐trifluoropentane‐2,4‐dione or 1,1,1‐trifluoropentane‐2,4‐dione and/or 1‐phenylbutane‐1,3‐dione as β‐diketone moiety. Standard methods were used to confirm the structure of complexes 1–6. X‐ray crystal structure analysis characterized complex 1 containing the ligand ethane‐1,2‐diamine and pentane‐2,4‐dione. The interactions of complexes 1–6 with calf thymus DNA (ct‐DNA) were followed by electronic absorption and fluorescence spectroscopy methods and by viscosity measurements. In contrast, interaction with Salmon Sperm DNA was investigated using the electrophoretic mobility shift assay. The results indicate a moderate affinity of complexes 1–6 for binding to DNA. Gel electrophoresis also shows that the studied complexes have a concentration‐dependent interaction with DNA. Spectroscopic fluorescence techniques were used to monitor the affinity of the complexes for bovine serum albumin (BSA). Complexes 1–6 showed satisfactory binding ability for BSA. Cytotoxicity analyses were performed on the human colorectal carcinoma HCT‐116 and healthy lung fibroblast MRC‐5 cell lines, showing that complex 5 exhibited selectivity between cancer and normal cells, which is critical for drug development.

Assessment of asphaltene and resin fractions in crude oil using laser-induced fluorescence spectroscopy based on modified Beer-Lambert (LIFS-MBL)

Crude oil is one of the most significant petrogenic sources of polycyclic aromatic compounds (PACs). These substances play an essential role in the pollution of the marine environment. Therefore, the rapid identification of this pollutant source and its fractions is vital. For this purpose, a fast and on-site method of laser-induced fluorescence spectroscopy based on modified Beer-Lambert (LIFS-MBL) is proposed here using solvent densitometry. Three optical parameters of the self-quenching (K), the extinction (α), and the peak concentration (Cp) are experimentally extracted from MBL graphs. Note that the parameters above are known to be unique characteristics of various crude oils. The corresponding compounds are generally classified into saturate, aromatic, resin, and asphaltene fractions, abbreviated as SARA. Differentiation among these fractions is achieved using the LIFS-MBL method by selecting the optimal excitation wavelength at 405 nm. This line effectively rules out the light aromatic rings and focuses on heavy fractions. The correlation of optical parameters with heavy oil fractions is verified according to analysis of variance. Statistical relations are proposed to calculate crude oil fractions values. The values of light fractions including saturate and aromatic components can also be determined by the heavy fractions. In this method, the test time is notably reduced from four days using the standard methods to less than half an hour according to the presented LIFS-MBL technique.

Excitation-emission fluorescence spectroscopy and sparse chemometric methods for grape seed oil classification and authentication

Grape seed oil (GSO) authentication and classification have recently become the most important aspects of edible oil quality control due to its high nutritional values. In the present contribution, excitation-emission fluorescence spectroscopy and sparse chemometric methods were used for the classification of GSO obtained from different Iranian grape genotypes and the detection of GSO adulteration with refined sunflower oil (SFO). The fluorescence spectra were collected in the wavelength ranges of λex = 200–500 nm and λem = 200–800 nm. To develop multivariate models, more than 200 samples from five different varieties of GSO were used. The sparse version of N-way partial least squares discriminant analysis (sNPLS-DA) was utilized to build an interpretable classification model. Classification accuracies for the sNPLS-DA model were 1.00 for predicting all grape genotypes. The collected fluorescence data revealed a distinct intensity difference between the Chafteh GSO and other GSOs in λex = 270–310 nm, and λem = 300–350 nm. To simulate the adulteration of GSO with SFO, 35 binary blends of Chafteh GSO samples (as a higher-quality oil) were prepared at adulterant levels from 10% to 50%. Sparse multivariate regression techniques, including the least absolute shrinkage and selection operator (Lasso), Ridge, and Elastic net were applied for quantitative analysis of GSO adulteration. The coefficient of multiple determination (R2) and the RMSE for the Lasso model were 0.914 and 0.013, respectively, for the external test sets. The results showed that the sparse chemometric methods coupled with excitation-emission fluorescence spectroscopy can be used as interpretable and reliable tools for quality control and adulteration detection of oil samples in the food industry and commerce.

Effect of isomorphic replacement of palygorskite on its heavy metal adsorption performance

Isomorphism is a common phenomenon in clay minerals, which is one of the main reasons for the change of mineral chemical composition and physical and chemical properties. In this study, four kinds of palygorskite with different degrees of isomorphic replacement of octahedral cations were selected. The microstructure and surface-interface properties of different Mg2+/Al3+ palygorskite were analyzed by X-ray diffraction, infrared spectroscopy, N2 adsorption-desorption method, X-ray fluorescence spectroscopy and other characterization methods, and the effects of isomorphic replacement on microstructure such as Mg content and surface-interface properties such as hydroxyl concentration and acid content were discussed. Taking Cu2+ as the target pollutant, the adsorption capacity of different palygorskite to Cu2+ was measured and discussed. Combining the surface hydroxyl concentration and acid amount of palygorskite, as well as the concentration, time and temperature of the reaction system, the rule of adsorption of the four palygorskite for Cu2+ was Mg-rich palygorskite (a-ATP) > palygorskite with equal Mg and Al (c-ATP) > Al-rich palygorskite (d-ATP) > palygorskite with the most Mg (b-ATP), demonstrating that when Mg was partially replaced by Al, palygorskite contained appropriate hydroxyl concentration and acid amount, resulting in the best adsorption effect for Cu2+ (29.2 mg/g). It is of great significance for resource utilization of palygorskite to reveal the regularity of heavy metal adsorption by isomorphic replacement of palygorskite.

The Kβ/Kα X-ray intensity ratios of Cu and Ag in Cu–Ag thin alloy films

Kβ/Kα X-ray intensity ratio values of Cu and Ag in CuxAg1-x (x = 53.5, 67.6, 76.9, 85.4, 89.6) thin alloy films produced from two different sources by the physical vapour deposition method have been determined by the Energy Dispersive X-ray Fluorescence Spectroscopy (EDXRF) method. The changes in the crystal structures of CuxAg1-x thin alloy films caused by the changes in the Cu and Ag concentrations, which affect the valence electronic structure have been investigated by the X-ray Diffraction (XRD) techniques. The obtained values were compared with the theoretical and fitted values for pure Cu and Ag elements.

Effect of Trace Metal Ions on the Conformational Stability of the Visual Photoreceptor Rhodopsin.

Trace metals are essential elements that play key roles in a number of biochemical processes governing human visual physiology in health and disease. Several trace metals, such as zinc, have been shown to play important roles in the visual phototransduction process. In spite of this, there has been little research conducted on the direct effect of trace metal elements on the visual photoreceptor rhodopsin. In the current study, we have determined the effect of several metal ions, such as iron, copper, chromium, manganese, and nickel, on the conformational stability of rhodopsin. To this aim, we analyzed, by means of UV-visible and fluorescence spectroscopic methods, the effects of these trace elements on the thermal stability of dark rhodopsin, the stability of its active Metarhodopsin II conformation, and its chromophore regeneration. Our results show that copper prevented rhodopsin regeneration and slowed down the retinal release process after illumination. In turn, Fe3+, but not Fe2+, increased the thermal stability of the dark inactive conformation of rhodopsin, whereas copper ions markedly decreased it. These findings stress the important role of trace metals in retinal physiology at the photoreceptor level and may be useful for the development of novel therapeutic strategies to treat retinal disease.

Time-resolved burst variance analysis

Quantifying biomolecular dynamics has become a major task of single-molecule fluorescence spectroscopy methods. In single-molecule Förster resonance energy transfer (smFRET), kinetic information is extracted from the stream of photons emitted by attached donor and acceptor fluorophores. Here, we describe a time-resolved version of burst variance analysis that can quantify kinetic rates at microsecond to millisecond timescales in smFRET experiments of diffusing molecules. Bursts are partitioned into segments with a fixed number of photons. The FRET variance is computed from these segments and compared with the variance expected from shot noise. By systematically varying the segment size, dynamics at different timescales can be captured. We provide a theoretical framework to extract kinetic rates from the decay of the FRET variance with increasing segment size. Compared to other methods such as filtered fluorescence correlation spectroscopy, recurrence analysis of single particles, and two-dimensional lifetime correlation spectroscopy, fewer photons are needed to obtain reliable timescale estimates, which reduces the required measurement time.

Synthesis, physicochemical characterisation and DNA binding study of a novel azo Schiff base Ni(II) complex

The azo Schiff base ligand was synthesised, along with its Ni(II) complex, by diazotisation of salicylaldehyde with 4-nitroaniline in accordance with the accepted literature approach. Using a variety of spectroscopic techniques, the resulting complex is analysed both quantitatively and qualitatively (Elemental analysis, FT-IR spectroscopy, UV-VIS spectroscopy, 1H NMR, etc.). Spectral measurements of the complex revealed a mole ratio of 1:1. The non-electrolytic nature of the complex is confirmed by molar conductance investigation. The unique azo compound had a tetrahedral shape as a result of the tetra coordination of two phenolic oxygen and two imine nitrogen. The ability of the metal complexes to bind DNA was examined using absorption spectroscopy, fluorescence spectroscopy, viscosity tests, and thermal denaturation methods. Experimental research suggests that complexes bind to DNA through intercalation.

Bodipy−corrole tetrads as heavy−atom−free triplet photosensitizers: Study of the energy transfer and application in triplet−triplet annihilation upconversion

Free base corrole can generate triplet state without invoking of heavy−atom effect, however, its weak and narrow absorption band in the visible spectral range make corrole not an ideal triplet photosensitizer. Herein, novel Bodipy−corrole tetrads were prepared by attaching visible light-harvesting antennae of Bodipy or electron spin converter with fullerene units (2B−Tur−Cor and B−C60−Tur−Cor) to solve the above problems, which show a stronger and broader absorption band in the visible region, and the maximum molar absorption coefficient is up to 2.58 × 105 M−1 cm−1 at 504 nm for 2B−Tur−Cor. By using of fluorescence spectroscopy, steady-state and time-resolved transient absorption spectroscopic methods, the singlet energy transfer progress from the Bodipy moiety to the corrole moiety was confirmed. Through fluorescence lifetimes and fluorescence excitation spectrum study, the rate constant of the singlet energy transfer was calculated as 9.58 × 107 s−1 for 2B−Tur−Cor and 2.45 × 108 s−1 for B−C60−Tur−Cor, respectively. The TD−DFT computation suggests that the first and second triplet state is localized in the corrole moiety and the Bodipy moiety for 2B−Tur−Cor, respectively, and the triplet state energy was calculated to be 1.37 eV and 1.61 eV, respectively. Nanosecond time−resolved transient difference absorption spectra show that the triplet excited states of these tetrads are localized on corrole unit, the lifetime of 2B−T−Cor is up to 143 μs, however, the lifetime of B−C60−Tur−Cor is only 3.3 μs. Long-lived triplet excited states play important role in applications of the triplet photosensitizers in photocatalysis or other photophysical processes concerning triplet–triplet−energy−transfer. As a proof of concept, these tetrads were used as heavy−atom−free organic triplet photosensitizers for triplet−triplet annihilation based upconversion. The upconversion quantum yield was up to 5.86% for 2B−Tur−Cor, however, it is only 0.52% for B−C60−Tur−Cor.

Oil species identification based on fluorescence excitation-emission matrix and transformer-based deep learning

After oil spills are found at sea, the identification on oil species can help determine the source of leakage and form the plan of post-accident treatment. Since the fluorometric characteristics of petroleum hydrocarbon reflect its molecular structure, the composition of oil spills could potentially be inferred using the fluorescence spectroscopy method. The excitation-emission matrix (EEM) includes additional fluorescence information in the dimension of excitation wavelength, which could be useful to identify oil species. This study proposed an oil species identification model using transformer network. The EEMs of oil pollutants are reconstructed into sequenced patch input that consists of the fluorometric spectra obtained under the different excitation wavelengths. The comparative experiments show that the proposed model can reduce the incorrect predictions and achieve higher identification accuracies than the regular convolutional neural networks that have been used in the previous studies. According to the structure of transformer network, an ablation experiment is also designed to evaluate the contributions of different input patches and seek for the optimal excitation wavelengths for oil species identification. The proposed model is expected to identify oil species, and even other fluorescent materials, based on the fluorometric spectra collected under multiple excitation wavelengths.

Size-dependent solvatochromism of CdS nanoparticles: Experimental absorption and fluorescence analyses

Owing to the quantum confinement effect, a nanocrystal shows a unique blend of molecular and solid properties: discrete energy levels with large polarizabilities which could respond to solvent environments. In this paper, we used steady-state fluorescence and absorption spectroscopy to study the size-dependent solvatochromism of CdS nanoparticles. The dipolar character of the ground state and the first excited state of the nanoparticle were also investigated with in the framework of Onsager reaction field theory using Ooshika–Mataga–Lippert equation. The results of the present study revealed that the solvatochromic responses were size dependent. Additionally, the responses increased with the particle size up to a certain size beyond which the response started to fall again. Hence, fine-tuning the nanoparticle's solvatochromic response and band gap is possible by appropriate choice of size. Moreover, size-dependent dipole moment values of CdS nanoparticles were determined. Based on the findings of the present study it was noted that steady-state fluorescence and absorption spectroscopy methods can efficiently be used to study the size-dependent solvatochromism of CdS nanoparticles.

Deconvolution of the fluorescence spectra measured through a needle probe to assess the functional state of the liver.

Currently, one of the most pressing issues for surgeons in the treatment of obstructive jaundice is the ability to assess the functional state of the liver and to detect and determine the degree of liver failure in a timely manner with simple and objective techniques. In this regard, the use of fluorescence spectroscopy method can be considered as one of the ways to improve the informativity of existing diagnostic algorithms in clinical practice and to introduce new diagnostic tools. Thus, the aim of the work was to study in vivo the functional state of liver parenchyma by the method of fluorescence spectroscopy implemented through a needle probe and assess the contribution of the main tissue fluorophores to reveal new diagnostic criteria. We compared data from 20 patients diagnosed with obstructive jaundice and 11 patients without this syndrome. Measurements were performed using a fluorescence spectroscopy method at excitation wavelengths of 365 and 450 nm. Data were collected using a 1 mm fiber optic needle probe. The analysis was based on the comparison of the results of deconvolution with the combinations of Gaussian curves reflecting the contribution of the pure fluorophores in the liver tissues. The results showed a statistically significant increase in the contribution of curves reflecting NAD(P)H fluorescence, bilirubin, and flavins in the group of patients with obstructive jaundice. This and the calculated redox ratio values indicated that the energy metabolism of the hepatocytes may have shifted to glycolysis due to hypoxia. An increase in vitamin A fluorescence was also observed. It may also serve as a marker of liver damage, indicating impaired vitamin A mobilization from the liver due to cholestasis. The results obtained reflect changes associated with shifts in the content of the main fluorophores characterizing hepatocyte dysfunction caused by accumulation of bilirubin and bile acids and after disturbance of oxygen utilization. The contributions of NAD(P)H, flavins, and bilirubin as well as vitamin A can be used for further studies as promising diagnostic and prognostic markers for the course of liver failure. Further work will include collecting fluorescence spectroscopy data in patients with different clinical effects of obstructive jaundice on postoperative clinical outcome after biliary decompression.

Quantitation of Tissue Amyloid via Fluorescence Spectroscopy Using Controlled Concentrations of Thioflavin-S.

Amyloids are misfolded proteins that aggregate into fibrillar structures, the accumulation of which is associated with the pathogenesis of many neurodegenerative diseases, such as Alzheimer's disease (AD). Early, sensitive detection of these misfolded aggregates is of great interest to the field, as amyloid deposition begins well before the presentation of clinical symptoms. Thioflavin-S (ThS) is a fluorescent probe commonly used to detect amyloid pathology. Protocols for ThS staining vary, but they often use high staining concentrations followed by differentiation, which causes varying levels of non-specific staining and potentially leaves more subtle amyloid deposition unidentified. In this study, we developed an optimized ThS staining protocol for the sensitive detection of β-amyloids in the widely used 5xFAD Alzheimer's mouse model. Controlled dye concentrations together with fluorescence spectroscopy and advanced analytical methods enabled not only the visualization of plaque pathology, but also the detection of subtle and widespread protein misfolding throughout the 5xFAD white matter and greater parenchyma. Together, these findings demonstrate the efficacy of a controlled ThS staining protocol and highlight the potential use of ThS for the detection of protein misfolding that precedes clinical manifestation of disease.

Polymeric Micelles Formulation of Combretastatin Derivatives with Enhanced Solubility, Cytostatic Activity and Selectivity against Cancer Cells.

Combretastatin derivatives is a promising class of antitumor agents, tubulin assembly inhibitors. However, due to poor solubility and insufficient selectivity to tumor cells, we believe, their therapeutic potential has not been fully realized yet. This paper describes polymeric micelles based on chitosan (a polycation that causes pH and thermosensitivity of micelles) and fatty acids (stearic, lipoic, oleic and mercaptoundecanoic), which were used as a carrier for a range of combretastatin derivatives and reference organic compounds, demonstrating otherwise impossible delivery to tumor cells, at the same time substantially reduced penetration into normal cells. Polymers containing sulfur atoms in hydrophobic tails form micelles with a zeta potential of about 30 mV, which increases to 40-45 mV when cytostatics are loaded. Polymers with tails of oleic and stearic acids form poorly charged micelles. The use of polymeric 400 nm micelles provides the dissolution of hydrophobic potential drug molecules. Micelles could significantly increase the selectivity of cytostatics against tumors, which has been shown using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, Fourier transform infrared (FTIR) spectroscopy, flow cytometry and fluorescence microscopy. Atomic force microscopy presented the difference between the unloaded micelles and those loaded with the drug: the size of the former was 30 nm on average, while the latter had a "disc-like" shape and a size of about 450 nm. The loading of drugs into the core of micelles was confirmed by UV and fluorescence spectroscopy methods; shifts of absorption and emission maxima into the long-wavelength region by tens of nm was observed. With FTIR spectroscopy, a high interaction efficiency of micelles with the drug on cells was demonstrated, but at the same time, selective absorption was observed: micellar cytostatics penetrate into A549 cancer cells 1.5-2 times better than the simple form of the drugs. Moreover, in normal HEK293T, the penetration of the drug is reduced. The proposed mechanism for reducing the accumulation of drugs in normal cells is the adsorption of micelles on the cell surface and the preservation of cytostatics to penetrate inside the cells. At the same time, in cancer cells, due to the structural features of the micelles, they penetrate inside, merging with the membrane and releasing the drug by pH- and glutathione-sensitive mechanisms. From a methodological point of view, we have proposed a powerful approach to the observation of micelles using a flow cytometer, which, in addition, allows us to quantify the cells that have absorbed/adsorbed cytostatic fluorophore and distinguish between specific and non-specific binding. Thus, we present polymeric micelles as drug delivery systems in tumors using the example of combretastatin derivatives and model fluorophore-cytostatic rhodamine 6G.

A self-organizing map approach to the analysis of lake DOM fluorescence for differentiation of organic matter sources.

The sources and properties of dissolved organic matter (DOM) in two lakes with different non-point source inputs were investigated by combining conventional three-dimensional fluorescence spectroscopy methods with a self-organizing map (SOM). To assess the DOM humification level, the representative neurons 1, 11, 25, and 36 were assessed. The SOM model showed that the DOM humification level of the Gaotang Lake (GT) which has a mainly agricultural non-point source input was significantly higher than that of the Yaogao Reservoir (YG) which has a mainly terrestrial source input (P < 0.01). The GT DOM mainly came from factors such as agricultural-related farm compost and decaying plants, while the YG DOM originated from human activities around the lake. The source characteristics of the YG DOM are obvious, with a high level of biological activity. Five representative areas in the fluorescence regional integral (FRI) were compared. The comparison showed that during the flat water period, the GT water column showed more terrestrial characteristics, even though the humus-like fractions in the DOM of both lakes were derived from microbial decomposition. Principal component analysis (PCA) showed that the agricultural lake water DOM (GT) was dominated by humus components, while the urban lake water DOM (YG) was dominated by authigenic sources.

Enhancement of Fluorescence Mediated by Silver Nanoparticles: Implications for Cell Imaging.

In this study, we report the surface enhanced fluorescence (SEF) of a biologically important organic dye, fluorescein (FL), by silver nanoparticles (Ag NPs) in an aqueous medium and its implications for human cell imaging. The as-synthesized Ag NPs were characterized by dynamic light scattering (DLS), zeta potential, transmission electron microscopy (TEM), and UV-vis absorption spectroscopic studies. The interaction and aggregation of FL dye with Ag NPs and a cationic surfactant, namely, cetyltrimethylammonium bromide (CTAB), were explored by UV-vis absorption and steady-state and time-resolved fluorescence spectroscopic methods. The distance-dependent fluorescence enhancement of FL due to Ag NPs in the solution was also theoretically correlated by three-dimensional finite-difference time-domain (3D-FDTD) simulation. The plasmonic coupling between neighboring NPs facilitated the augmentation of the local electric field, thereby producing various "hotspots" that influence the overall fluorescence of the emitter. J-type aggregates of FL in the presence of the CTAB micelles and Ag NP mixed solution were confirmed by electronic spectroscopy. The density functional theoretical (DFT) study revealed the electronic energy levels associated with different forms of FL dye in the aqueous solution. Most interestingly, the Ag NP/FL mixed system used in fluorescence imaging of human lung fibroblast cells (WI 38 cell line) showed a significantly stronger green fluorescence signal compared to that of FL after an incubation period of only 3 h. This study confirms that the Ag NP mediated SEF phenomenon of the FL dye is also manifested in the intracellular medium of human cells giving a brighter and more intense fluorescence image. The cell viability test after exposure to the Ag NP/FL mixed system was confirmed by the MTT assay method. The proposed study may have an implication as an alternate approach for human cell imaging with higher resolution and more contrast.