Total Fluorescence Research Articles

Deep understanding of LiCoO2 electrode degradation for optimized recycling strategies

Empowered by a synergistic combination approach of Scanning Transmission X-ray Microscopy (STXM) imaging and X-ray Absorption Near Edge Structure (XANES) spectroscopy, a quantitative analysis on the composition and spatial distribution of the cathode in a failed 18650 LiCoO2 (LCO) battery was conducted. Distinct compositional differences between the central and peripheral cathode regions in the failed LCO battery were discerned by utilizing bulk XANES in Total Electron Yield (TEY) and Fluorescence Yield (FY) modes. The central region shows more severe degradation in terms of LCO reduction and excess cathode-electrolyte interface (CEI) buildup. Meanwhile, the STXM technique precisely imaged a specific region of interest in the center of the cathode, covering C, O, and F K-edges, and Co L2,3-edge, to deeply investigate the degradation. Quantitative chemical mapping with spatially resolved XANES spectroscopy, facilitate an in-depth understating of the interfacial reactions on battery electrodes and battery failure fundamentals.

Continuous distribution of cancer cells in the cell cycle unveiled by AI-segmented imaging of 37,000 HeLa FUCCI cells

Classification of live or fixed cells based on their unlabeled microscopic images would be a powerful tool for cell biology and pathology. For such software, the first step is the generation of a ground truth database that can be used for training and testing AI classification algorithms. The Application of cells expressing fluorescent reporter proteins allows the building of ground truth datasets in a straightforward way. In this study, we present an automated imaging pipeline utilizing the Cellpose algorithm for the precise cell segmentation and measurement of fluorescent cellular intensities across multiple channels. We analyzed the cell cycle of HeLa–FUCCI cells expressing fluorescent red and green reporter proteins at various levels depending on the cell cycle state. To build the dataset, 37,000 fixed cells were automatically scanned using a standard motorized microscope, capturing phase contrast and fluorescent red/green images. The fluorescent pixel intensity of each cell was integrated to calculate the total fluorescence of cells based on cell segmentation in the phase contrast channel. It resulted in a precise intensity value for each cell in both channels. Furthermore, we conducted a comparative analysis of Cellpose 1.0 and Cellpose 2.0 in cell segmentation performance. Cellpose 2.0 demonstrated notable improvements, achieving a significantly reduced false positive rate of 2.7 % and 1.4 % false negative. The cellular fluorescence was visualized in a 2D plot (map) based on the red and green intensities of the FUCCI construct revealing the continuous distribution of cells in the cell cycle. This 2D map enables the selection and potential isolation of single cells in a specific phase. In the corresponding heatmap, two clusters appeared representing cells in the red and green states. Our pipeline allows the high-throughput and accurate measurement of cellular fluorescence providing extensive statistical information on thousands of cells with potential applications in developmental and cancer biology. Furthermore, our method can be used to build ground truth datasets automatically for training and testing AI cell classification. Our automated pipeline can be used to analyze thousands of cells within 2 h after putting the sample onto the microscope.

In vivo assessment of bladder cancer with diffuse reflectance and fluorescence spectroscopy: A comparative study.

The aim of this work is to assess the performance of multimodal spectroscopic approach combined with single core optical fiber for detection of bladder cancer during surgery in vivo. Multimodal approach combines diffuse reflectance spectroscopy (DRS), fluorescence spectroscopy in the visible (405 nm excitation) and near-infrared (NIR) (690 nm excitation) ranges, and high-wavenumber Raman spectroscopy. All four spectroscopic methods were combined in a single setup. For 21 patients with suspected bladder cancer or during control cystoscopy optical spectra of bladder cancer, healthy bladder wall tissue and/or scars were measured. Classification of cancerous and healthy bladder tissue was performed using machine learning methods. Statistically significant differences in relative total haemoglobin content, oxygenation, scattering, and visible fluorescence intensity were found between tumor and normal tissues. The combination of DRS and visible fluorescence spectroscopy allowed detecting cancerous tissue with sensitivity and specificity of 78% and 91%, respectively. The addition of features extracted from NIR fluorescence and Raman spectra did not improve the quality of classification. This study demonstrates that multimodal spectroscopic approach allows increasing sensitivity and specificity of bladder cancer detection in vivo. The developed approach does not require special probes and can be used with single-core optical fibers applied for laser surgery.

Ligand-Free Silver Nanoparticles: An Innovative Strategy against Viruses and Bacteria.

The spread of antibiotic-resistant bacteria and the rise of emerging and re-emerging viruses in recent years constitute significant public health problems. Therefore, it is necessary to develop new antimicrobial strategies to overcome these challenges. Herein, we describe an innovative method to synthesize ligand-free silver nanoparticles by Pulsed Laser Ablation in Liquid (PLAL-AgNPs). Thus produced, nanoparticles were characterized by total X-ray fluorescence, zeta potential analysis, transmission electron microscopy (TEM), and nanoparticle tracking analysis (NTA). A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to evaluate the nanoparticles' cytotoxicity. Their potential was evaluated against the enveloped herpes simplex virus type 1 (HSV-1) and the naked poliovirus type 1 (PV-1) by plaque reduction assays and confirmed by real-time PCR and fluorescence microscopy, showing that nanoparticles interfered with the early stage of infection. Their action was also examined against different bacteria. We observed that the PLAL-AgNPs exerted a strong effect against both methicillin-resistant Staphylococcus aureus (S. aureus MRSA) and Escherichia coli (E. coli) producing extended-spectrum β-lactamase (ESBL). In detail, the PLAL-AgNPs exhibited a bacteriostatic action against S. aureus and a bactericidal activity against E. coli. Finally, we proved that the PLAL-AgNPs were able to inhibit/degrade the biofilm of S. aureus and E. coli.

Levels of rare earth elements on three abandoned mining sites of bauxite in southern Italy: A comparison between TXRF and ICP-MS

The essential utilization of rare earth elements (REEs) for the production of several electronic devices is making the demand for them being increased all the time. This extensive use of these elements has also increased concern about human and environmental health. Previous studies have shown that REE levels are higher in environmental samples near mining sites, and they are highly possible to be transferred to biota. In this study, REE levels were determined in environmental samples collected from three abandoned mining sites of bauxite (Gargano, Otranto, and Spinazzola) in the region of Puglia, Southern Italy. The samples were digested and analyzed by two different techniques, Total X-Ray Fluorescence (TXRF) and Inductively Coupled Plasma – Mass Spectroscopy (ICP-MS) to investigate which technique is the most suitable for analysis of the REE content in samples from abandoned mining sites of bauxite. Only 6 REEs could be detected by TXRF, while all REEs were detected in all the samples by ICP-MS. Spinazzola is the richest site and Ce the most abundant REE in all three regions. REE levels are correlated between the soil and biota samples in many cases, although the calculation of the bioconcentration factor showed that REEs are not bioaccumulative. ICP-MS seems to be a more suitable technique for analysis of the whole REE content in environmental samples from abandoned mining sites of bauxite.

Controlled dual activity of an organometallic antibiotic through micelle formulation

Sodium Lauryl Sulphate (SLS) and Cetyltrimethylammonium bromide (CTAB) were employed to create micelles (SLS@CIPTIN and CTAB@CIPTIN) of the organometallic antibiotic Ph2Sn(CIP)2 (CIPTIN) (where HCIP= ciprofloxacin). The characterization of the micelles was conducted in the solid-state using melting point, Attenuated Total Reflectance-Fourier Transform Infra-Red (ATR-FTIR), 119Sn Mössbauer and X-ray Fluorescence (XRF) spectroscopies, and Thermogravimetry/Differential Thermal Analysis (TG-DTA), while in solution, by Ultra violet-Visible (UV–Vis) and Nucleus Magnetic Resonance (1HNMR ) spectroscopic techniques were employed. The compounds were tested for their antiproliferative activity against human breast adenocarcinoma cells: MCF-7 (hormone depended (HD)) and MDA-MB-231 (hormone independent (HI)). The in vitro and in vivo non-genotoxicity was confirmed with micronucleus (MN) and Artemia salina assay. The antiproliferative mechanism of action of SLS@CIPTIN and CTAB@CIPTIN was studied by cell morphology and cell cycle arrest. The antibacterial activity was evaluated using Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Inhibition Zones (IZ) against Minimum Bactericidal Concentration (MBC) against Gram-negative strains P. aeruginosa, E. coli, as well as Gram-positive strains S. epidermidis and S. aureus. The results indicate that both the investigated CIPTIN and its supramolecular assembly (SLS@CIPTIN) exhibit activity. Therefore, the encapsulation of antimicrobial compounds can indeed be regarded as an attractive strategy for the development of new dual-active agents. These agents offer concurrent anticancer and antibacterial attributes without inducing toxicity.

Anthropogenic dissolved organic matter accumulation fuels greenhouse gas diffusive emissions in urban lakes along trophic state levels

Lakes are an important reservoir of dissolved organic matter (DOM) and also emit large amounts of greenhouse gases (GHGs), such as carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O). Yet, the linkages between quantity and quality of DOM and GHGs emissions are still not well understood. Hence, we investigated how DOM optical properties mediated dissolved GHGs levels and diffusive fluxes along a trophic state level in subtropical urban lakes, China. Our study showed that anthropogenic protein-like DOM (tryptophan- and tyrosine-like components) dominated the DOM pools, comprising 62% of total fluorescence DOM (FDOM). Partial least squares path modeling (PLS-PM) showed that the direct contribution of relative abundance of DOM to CO2 and CH4 diffusion in the highly-eutrophic state was higher than in the mesotrophic and lightly/moderately-eutrophic states. In particular, protein-like DOM primarily accounted for CO2 variations (44%-49%), while humic- and protein-like DOM contributed comparably to CH4 variations (38%-40% vs. 36%-40%). Furthermore, in the lightly/moderately- and highly-eutrophic states, nutrients contributed to DOM, CO2 and CH4 in both direct and indirect ways, while phytoplankton (chlorophyll a, Chl-a) negatively and directly affected carbon emissions. However, only nitrogen content directly contributed to N2O production and diffusion in the mesotrophic and lightly/moderately-eutrophic states. We concluded that protein-like DOM enrichment had the potential to trigger CO2 and CH4 diffusion and this process was largely mediated by nutrient levels. N2O diffusion was found to be mainly influenced by nitrogen loadings and forms. Our findings highlight the urgency for effective strategies to reduce anthropogenic DOM inputs and control nutrient levels to reduce GHGs diffusion in highly human-impacted aquatic systems.

Characterization and function of particulate organic matter: Evidence from lakes undergoing ecological restoration

Particulate organic matter (POM) plays a crucial role in the organic composition of lakes; however, its characteristics remain poorly understood. This study aimed to characterize the structure and composition of POM in Lake Baiyangdian using many kinds of techniques and investigate the effects of different extracted forms of POM on water quality. The suspended particulate matter in the lake had complex compositions, with its components primarily derived from aquatic plants and their detritus. The organic matter content of the suspended particulate matter was relatively high (organic carbon content 27.29–145.94 g/kg) for the sum of three extractable states (water-extracted organic matter [WEOM], humic acid, and fulvic acid) and one stable bound state (humin). Spatial distribution analysis revealed that the POM content in the water increased from west to east, which was consistent with the water flow pattern influenced by the Baiyangdian water diversion project. Fluorescence spectroscopy analysis of the WEOM showed three prominent peaks with excitation/emission wavelengths similar to those of dissolved organic matter peaks. These peaks were potentially initial products of POM conversion into dissolved organic matter. Furthermore, the intensity of the WEOM fluorescence peak (total fluorescence peak intensity) was negatively correlated with the inorganic nitrogen concentration in water (p < 0.01), while the intensity of the HA fluorescence peak showed a positive correlation with the inorganic nitrogen concentration (p < 0.01). This suggested that exogenous organic matter inputs led to the diffusion of alkaline dissolved nitrogen from sediment into water, while degradation processes of aquatic plant debris contributed to the decrease in inorganic nitrogen concentrations in the water column. These findings enhance our understanding of POM characteristics in shallow lakes and the role of POM in shallow lake ecosystems.

Seasonal changes in water-soluble brown carbon (BrC) at Nanling background station in South China

Brown carbon (BrC) is an important light-absorbing component of organic carbon (OC), causing large uncertainty in aerosol radiative forcing evaluation and being related to health issues as well. Knowledge of BrC in an atmospheric background station is beneficial to understand its role in a changing climate. A year-long sampling campaign was conducted at Nanling background station to get a comprehensive knowledge of WS-BrC, a total of seventy-two PM2.5 samples throughout a year were used. Light absorption and fluorescence spectra of WSOC were analyzed synchronously using a fluorescence spectrophotometer. The low levels of PM2.5, OC, and elemental carbon (EC) conferred a background site. The optical properties of WS-BrC were characterized using excitation-emission matrix (EEM) fluorescence spectroscopy. The WS-BrC made a significant contribution (365 nm, 18% ± 10%) to total carbonaceous aerosol absorption. The mass absorption efficiency (MAE) of WS-BrC is 0.81 ± 0.34 m2 gC–1, and varies among seasons due to the different sources or atmospheric processing. Three EEM fluorescent components were identified by parallel factor (PAFAFAC) analysis, including two humic-like substances (HULIS, C1, C2), and one phenolic-like component. The HULIS components accounted for approximately 70% of the total fluorescence intensities. Primary combustion emissions showed enhanced activity during the winter and spring seasons, but there were no significant influences on WS-BrC in spring. Secondary sources contributed significantly to WS-BrC during winter, summer, and autumn (all exceeding 50%), except for spring. Photooxidation is a significant process in the formation of secondary WS-BrC in winter and autumn, but there may be another formation pathway in summer, i.e., the ammonia pathway. This study contributes to our understanding of BrC in the background atmosphere.

A versatile beamline for soft x-ray reflectivity, absorption, and fluorescence measurements at Indus-2 synchrotron source.

A versatile beamline for performing reflectivity, fluorescence, and absorption experiments in the soft x-ray region of 100-1500eV is commissioned on a bending magnet port of the Indus-2 synchrotron source. A high vacuum 2-axis reflectometer with x, y, and z sample scanning stages is installed. This reflectometer is used to measure the reflectivity of large samples up to 300mm in length and 5kg in weight. This feature is useful for characterizing x-ray optical elements, such as mirrors, gratings, and multilayers. A flange mounted silicon drift detector is installed in the downstream of the reflectometer for soft x-ray fluorescence measurements. The soft x-ray absorption measurements are carried out in the total electron yield and partial fluorescence yield modes. Integration of three different experimental techniques in the experimental station makes the beamline versatile for materials science applications as it provides structural, chemical, and electronic state information by performing the required experiments in an identical environment. The beamline uses a varied line spacing plane grating monochromator and gives a high flux (∼109 to 1011 photon/s) with a moderate resolution (λ/Δλ ~1000-5000). A three-mirror-based higher harmonic setup is incorporated to get rid of harmonics and to get a high spectral purity monochromatic beam with less than 0.1% harmonic content. In the present article, the beamline optical scheme, mechanical configuration, and details of the experimental setups are presented, along with a few representative results of each experimental mode.

Origin of microscopically coupled ferromagnetic Cu-ions in a distorted system of Cu-doped ZnO and their synchrotron-based electronic structures

Spintronics-based studies have produced significant attention in the last decade while claiming the observation of room temperature ferromagnetism (RTFM). Nevertheless, there is a lack of consensus on a mechanism responsible for this phenomenon. In this study, we focus on Cu-doped ZnO (ZCO) to understand the microscopic origin of RTFM and the role of different oxidation states of Cu in RTFM. We have performed different spectroscopic techniques using synchrotron facilities. The values of spin-moment obtained from x-ray magnetic circular dichroism sum-rule truly exhibit a ferromagnetic interaction in the nanocrystalline powder of ZCO with ∼0.58 μB for 5% of Cu concentration in the total fluorescence yield mode. Such an enhanced magnetization is attributed to the presence of Cu2+, which is mainly localized in the bulk region. Cu in ZCO is mostly dominated by the presence of Cu2+. This is clearly reflected by the profiles of x-ray photoemission spectroscopy. Consequently, the weakly magnetized total electron yield mode is attributed to a state of magnetic frustration as the majority of Cu3+ is found on the surface. Some of these Cu3+ when come in the vicinity of Cu2+ ions result in a highly correlated state of double exchange mechanism, which is the microscopic origin of RTFM in ZCO. The coupling between Cu2+-Cu3+ is mediated via oxygen vacancies (VO), the presence of which is confirmed through the features of electron energy loss spectroscopy over different edges. The confirmation of VO is also supported by the deconvolution of E2high-phonon in the Raman spectra. Moreover, the defects in the local electronic structures of ZCO are demonstrated by the deconvoluted spectra of Cu L3 x-ray absorption spectroscopy. The images obtained from high-resolution transmission electron microscopy confirm the incorporation of Cu into the wurtzite crystal of ZnO. A clear enhancement in magnetization upon an increase in carriers of Cu in ZCO indicates carrier-induced ferromagnetism. Cu2+ and VO are the two attributes of RTFM in ZCO.

The effects of exogenous substances on the color of heated bones.

Burned bone coloration has been used for decades to help in the bioanthropological analysis of burned human bones. However, there is a variety of factors that can interfere with the coloration manifested by bones exposed to heat, resulting in colors that differ from the usual black to white gradient. In this study, we evaluated possible causes of unusual coloration changes and hues in burned bone. For that purpose, defleshed fresh pig (Sus scrofa) ribs as well as fresh and dry human clavicles were burned at four different temperatures (500, 700, 900 and 1100°C) in contact with different materials (CaO, Zn, Fe, Cu, Mn, and polyester cloth). Observable color changes were assessed through naked eye observation and description, Munsell color charts, and reflectance spectrophotometry. Additionally, chemical changes in bone were assessed using Fourier-transform infrared spectroscopy in attenuated total reflectance (FTIR-ATR) and x-ray fluorescence (XRF). Our results showed that some materials did affect usual burned bone coloration (Fe, Mn, Cu, Zn) and correspondent FTIR-ATR and XRF spectra. As for other materials, although no effect on visual bone coloration was observed, they still affected FTIR-ATR and XRF spectra (CaO and cloth). This study can contribute to the anthropological analysis of burned human remains, providing some answers to what can cause unusual types of heat-induced colorations.

Novel 40 µm spot size 3050/3200 nm DFG laser versus CO2 laser for laser-assisted drug delivery.

The use of ablative fractional lasers to enhance the delivery of topical drugs through the skin is known as laser-assisted drug delivery. Here, we compare a novel 3050/3200 nm difference frequency generation (DFG) fiber laser (spot size: 40 µm) to a commercially used CO2 laser (spot size: 120 µm). The objective is to determine whether differences in spot size and coagulation zone (CZ) thickness influence drug uptake. Fractional ablation was performed on ex-vivo human abdominal skin with the DFG (5 mJ) and CO2 (12 mJ) lasers to generate 680 µm deep lesions. To evaluate drug delivery, 30 kDa encapsulated fluorescent dye was topically applied to the skin and histologically analyzed at skin depths of 100, 140, 200, 400, and 600 µm. Additionally, transcutaneous permeation of encapsulated and 350 Da nonencapsulated dye was assessed using Franz Cells. The DFG laser generated smaller channels (diameter: 56.5 µm) with thinner CZs (thickness: 22.4 µm) than the CO2 laser (diameter: 75.9 µm, thickness: 66.8 µm). The DFG laser treated group exhibited significantly higher encapsulated dye total fluorescence intensities after 3 h compared to the CO2 laser treated group across all skin depths (p < 0.001). Permeation of nonencapsulated dye was also higher in the DFG laser treated group vs the CO2 laser treated group after 48 h (p < 0.0001), while encapsulated dye was not detected in any group. The DFG laser treated skin exhibited significantly higher total fluorescence uptake compared to the CO2 laser. Additionally, the smaller spot size and thinner CZ of the DFG laser could result in faster wound healing and reduced adverse effects while delivering similar or greater amount of topically applied drugs.

BBQ methods: streamlined workflows for bacterial burden quantification in infected cells by confocal microscopy.

Accurate quantification of bacterial burden within macrophages, termed bacterial burden quantification (BBQ), is crucial for understanding host-pathogen interactions. Various methods have been employed, each with strengths and weaknesses. This article addresses limitations in existing techniques and introduces two novel, automated methods for BBQ within macrophages based on confocal microscopy data analysis. The first method refines total fluorescence quantification by incorporating filtering steps to exclude uninfected cells, while the second method calculates total bacterial volume per cell to mitigate potential biases in fluorescence-based readouts. These workflows utilize PyImageJ and Cellpose software, providing reliable, unbiased, and rapid quantification of bacterial load. The proposed workflows were validated using Salmonella enterica serovar Typhimurium and Mycobacterium tuberculosis models, demonstrating their effectiveness in accurately assessing bacterial burden. These automated workflows offer valuable tools for studying bacterial interactions within host cells and provide insights for various research applications.

Unveiling angular sweep total fluorescence Spectroscopy:A novel multidimensional technique for analysis of complex multi-fluorophoric systems

Analyzing multi-fluorophoric real systems without pre-processing is challenging, often addressed with unconventional fluorescence techniques and chemometrics. In this context, we introduce a novel addition - ‘Angular Sweep Total Fluorescence Spectroscopy (ASTFS),' to the arsenal of multidimensional steady-state fluorescence spectroscopic techniques. ASTFS utilizes a series of variable angle synchronous fluorescence spectra, strategically covering the fluorescence region between the first and second-order Rayleigh scattering ridges. The plot features a minimal data matrix size, avoids Rayleigh scattering signals, and incurs no blind regions. The study delves into the instrumental configurations for spectral acquisition, highlights the enhanced spectral resolution due to the band-narrowing effect, and discusses other notable features of the ASTFS plot. Further, this technique is reported to be effective in analyzing analytes in complex systems with strong background fluorescence, such as milk. The antibiotic— Norfloxacin is quantified via minimal pre-processing in milk samples and yields excellent analytical figures of merit.

Tracking the Activity and Position of Mitochondrial β-Barrel Proteins.

Total interference reflection fluorescence (TIRF) microscopy of lipid bilayers is an effective technique for studying the lateral movement and ion channel activity of single integral membrane proteins. Here we describe how to integrate the mitochondrial outer membrane preprotein translocase TOM-CC and its β-barrel protein-conducting channel Tom40 into supported lipid bilayers to identify possible relationships between movement and channel activity. We propose that our approach can be readily applied to membrane protein channels where transient tethering to either membrane-proximal or intramembrane structures is accompanied by a change in channel permeation.

Inducing molecular orientation in solution-processed thin films of fluorene-bithiophene-based copolymer: thermal annealing vs. solvent additive.

A deep understanding of the factors influencing the morphology of thin films based on conjugated polymers is essential to boost their performance in optoelectronic devices. Herein, we investigated the electronic structure and morphology of thin films of the copolymer poly(9,9-dioctyl-fluorenyl-co-bithiophene) (F8T2) in its pristine form as well as samples processed with the solvent additive 1,8-diiodooctane (DIO) or post-processed through thermal annealing treatment. Measurements were carried out using angle-resolved S K-edge NEXAFS (near-edge X-ray absorption fine structure) in total electron yield (TEY) and fluorescence yield (FY) detection modes. Two main transitions were observed at the S 1s NEXAFS spectra: S 1s → π* and S 1s → σ* (S-C). The observed dichroism pointed to a face-on orientation of the conjugated backbone, which was significantly increased for F8T2 films processed with DIO. Resonant Auger decay spectra were obtained and analyzed using the core-hole clock (CHC) method. An enhancement in the charge transfer process was observed for thermally annealed films, especially for samples processed with DIO, corresponding to an increase in film ordering. Furthermore, the investigated films were characterized using X-ray photoelectron spectroscopy, attesting to the presence of the thiophene unit in the samples and demonstrating that some of its sulfur atoms were positively polarized in the F8T2 films. All these experimental findings were compared with molecular dynamics (MD) simulations of film evaporation with and without DIO. The use of MD, together with mathematical modeling, was able to explain the major effects found in the experiments, including the polarization of sulfur atoms. The simultaneous use of powerful spectroscopic techniques and theoretical methods shed light on key aspects linking film morphology with fabrication procedures.

Single-Molecule Approaches to Study DNA Condensation.

Proteins drive genome compartmentalization across different length scales. While the identities of these proteins have been well-studied, the physical mechanisms that drive genome organization have remained largely elusive. Studying these mechanisms is challenging owing to a lack of methodologies to parametrize physical models in cellular contexts. Furthermore, because of the complex, entangled, and dense nature of chromatin, conventional live imaging approaches often lack the spatial resolution to dissect these principles. In this chapter, we will describe how to image the interactions of λ-DNA with proteins under purified and cytoplasmic conditions. First, we will outline how to prepare biotinylated DNA, functionalize coverslips with biotin-conjugatedpoly-ethylene glycol (PEG), and assemble DNA microchannels compatible for the imaging of protein-DNA interactions using total internal fluorescence microscopy. Then we will describe experimental methods to image protein-DNA interactions in vitro and DNAloop extrusion using Xenopus laevis egg extracts.

Shedding light on the ageing of extra virgin olive oil: Probing the impact of temperature with fluorescence spectroscopy and machine learning techniques

This work systematically investigates the oxidation of extra virgin olive oil (EVOO) under accelerated storage conditions with UV absorption and total fluorescence spectroscopy. With the large amount of data collected, it proposes a method to monitor the oil's quality based on machine learning (ML) applied to highly-aggregated data.EVOO is a high-quality vegetable oil that has earned worldwide reputation for its numerous health benefits and excellent taste. Despite its outstanding quality, EVOO degrades over time due to oxidation, which can affect both its health qualities and flavour. Therefore, it is highly relevant to quantify the effects of oxidation on EVOO and develop methods to assess it that can be easily implemented under field conditions, rather than in specialized analytical laboratories.The ML approach indicates that the two excitation wavelengths (480 nm) and (300 nm) exhibit the maximum relative change in fluorescence intensity during the ageing for the majority of the oils, thus identifying the wavelengths which are more informative for quality prediction. Also, the paper proposes a method for the prediction of olive oil quality using highly-aggregated data. Such a method is of interest because it paves the way to the realization of a low-cost, portable device for in-field quality control.The following study demonstrates that fluorescence spectroscopy has the capability to monitor the effect of oxidation and assess the quality of EVOO, even when the data are highly aggregated. It shows that complex laboratory equipment is not necessary to exploit fluorescence spectroscopy using the proposed method and that cost-effective solutions, which can be used in-field by non-scientists, could provide an easily-accessible assessment of the quality of EVOO.

Ancient terminal water treatment methods still work: Removing micropollutants in drinking water by simultaneous boiling and VUV

Micropollutant in drinking water may not be effectively removed during centralized treatment. To reduce potential health risks, water treatment at the user terminal is warranted. This study investigated the performance of VUV coupled with boiling to simultaneously remove contaminants, a common household water treatment method in developing countries, for the treatment of micropollutants. Compared to individual boiling or VUV treatment, the VUV/BO process was more effective in improving the degradation of micropollutants. The results showed that heating promoted the direct photolysis and HO oxidation capabilities in the VUV process, with the HO generation increasing from 36.56 μM at 30 °C to 80.48 μM at 90 °C. At a reaction temperature of 100 °C, the degradation rates of malathion (MAL), dimethoate (DMT), trichloroacetic acid (TCAA), and dichloroacetic acid (DCAA) by VUV were increased by 0.81, 1.19, 1.34, and 0.90 times respectively compared to 25 °C. Analysis of the organic composition of actual water samples showed that the total fluorescence intensity of organic matter decreased significantly after VUV oxidation, mainly degrading humic-like substances. Using GC–MS analysis, this study identified 11 oxidative products of the DMT, and proposed two possible degradation pathways. Toxicity evaluation of the intermediate products of DMT demonstrated that they were less toxic than the parent contaminants. The results indicate that the heating-coupled water treatment process provides an effective technology for the treatment of micropollutants in terminal drinking water, and this process is worth promoting in the future.