Methods and Applications in Fluorescence荧光方法与应
Methods and Applications in Fluorescence(英文缩写 METHODS APPL FLUORES),ISSN 2050-6120,eISSN 2050-6120,中文译名:荧光方法与应 是一本学术期刊。本页汇总该期刊的最新影响因子、分区信息以及最新收录于 PubMed 的文献,帮助您快速了解期刊全貌。
发文量统计区间：2025-10-01 至 2026-09-30，按本站收录文献的发表日期统计。
期刊介绍
历年影响因子趋势
| JCR 数据年份 | 影响因子 | JCR 分区 |
|---|---|---|
| 2021 | 3.849 | Q2 |
| 2022 | 3.200 | Q2 |
| 2023 | 2.400 | Q3 |
| 2024 | 2.400 | Q3 |
| 2025 | 2.500 | Q3 |
Methods and Applications in Fluorescence 最新收录文献
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1. {"_":"Host-dependent structural and optical properties and upconversion ratiometric thermometry of Yb/Er-doped NaYF, NaGdF, NaScFnanophosphors.","sup":["3+","3+"],"sub":["4","4","4"]}
PMID:日期:2026-09-30This work presents a systematic study of the structural, morphological, and optical properties, along with the upconversion ratiometric thermometry of NaGdYbErF, NaYYbErF, NaYYbErF, and NaScYbErFnanophosphors. Structural analysis confirms the hexagonal-phase for all the nanophosphors except NaScYbErF. Upconversion emission studies show emission bands corresponding to theH→I,S→I, andF→Itransitions of Erions. Upconversion emission intensities and green-to-red emission intensity ratios strongly depend on Ybconcentration and phonon energy of the hosts. The temperature-dependent population distribution between the thermally coupled levels,HandS, was used for upconversion-based thermometry. The plot of temperature sensitivity as a function of absolute temperature shows a linear variation for all these nanophosphors. The NaScYbErFshows the lowest temperature sensitivity, possibly because of the higher phonon energy of this host. Thus, the findings of this work provide valuable insights into the role of the sensitizer and host matrix in designing efficient upconversion nanothermometers.
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2. A practical framework for measuring protein oligomerization equilibria by fluorescence correlation spectroscopy.
PMID:日期:2026-09-23Protein oligomerization equilibria are central to many biological processes and are often highly sensitive to environmental conditions such as ionic
strength, pH, and ligand binding. Quantitative characterization of these equilibria remains experimentally challenging because stable protein com-
plexes frequently dissociate only at concentrations that are difficult to access with conventional biophysical methods. Fluorescence correlation spectroscopy (FCS) is uniquely suited to this problem, as it provides direct access to diffusion coefficients of fluorescently labeled proteins at nanomolar concentrations. However, the quantitative interpretation of FCS data from oligomeric systems requires a rigorous mathematical framework and careful experimental practice that have not previously been described in sufficient detail to guide implementation. Here, we provide a comprehensive description of the experimental workflow and analytical framework for determining dissociation equilibrium constants by FCS, covering instrument calibration, sample preparation, data quality control, afterpulse correction, and nonlinear least-squares fitting. We discuss common sources of error and provide practical guidance on critical experimental considerations including surface passivation, buffer preparation, equilibration time, and the role of labeling efficiency. Using the homotrimeric sliding clamp PCNA as a model system, we demonstrate the complete workflow under a range of KCl concentrations and show that moderate ionic strength stabilizes the PCNA trimer while very high salt partially destabilizes the complex. The approach is general and applicable to any reversible protein self-association reaction accessible by fluorescence detection at low protein concentrations.
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3. {"_":"Revealing alkali ion modulation on the nucleation phase of highly concentrated-synuclein probed by Thioflavin T lasing spectroscopy.","i":["α"]}
PMID:日期:2026-09-23Early nucleation events during-synuclein (S) aggregation remain challenging to investigate because conventional fluorescence assays lose sensitivity in highly concentrated biomolecular environments. Here, we present Thioflavin T (ThT) lasing spectroscopy in a Fabry-Pérot microcavity as a sensitive methodological approach for probingS aggregation within condensate-like microenvironments that more closely mimic intracellular molecular crowding. Beyond conventional lasing-threshold analysis, we introduce the first automated frame-by-frame analysis of ThT lasing spectra using machine-vision algorithms, enabling statistical evaluation of individual lasing events throughout the aggregation process. As a demonstration of this methodology, we investigate how physiologically abundant monovalent and divalent cations (Na, K, Mg, and Ca) modulate the nucleation phase of highly concentratedS. Ion-specific differences were identified in both lasing-threshold kinetics and spectral evolution. Monovalent cations produced complex, non-monotonic threshold behavior accompanied by transient spectral shifts, consistent with incomplete electrostatic screening and dynamic accessibility of ThT to early oligomeric species. In contrast, divalent cations generated more stable lasing wavelengths and distinct threshold regimes, reflecting stronger electrostatic screening. These results demonstrate that automated analysis of ThT lasing provides complementary information beyond conventional fluorescence, enabling sensitive interrogation of nucleation processes in highly crowded protein systems. The presented methodology establishes lasing spectroscopy combined with machine-vision analysis as a powerful platform for investigating early protein aggregation under physiologically relevant condensed conditions.
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4. Fibre-enhanced fluorescent-based immunosorbent assay.
PMID:日期:2026-09-22During the manufacture of many bio-pharmaceutical products, it is difficult to monitor process yields in-line with production. This is particularly the case with egg-based influenza vaccine manufacture, where current assay methods, such as the fluorescent focus assay (for live-attenuated virus vaccines), require timescales incompatible with normal production cycles. This is due to the requirement to perform cell culture incubation as part of this assay, which takes several days to complete before a result is obtained. Other, more rapid assay methods, such as the enzyme-linked immunosorbent assay, rely on peroxidase-labelled antibodies, which are relatively expensive, and current plate-based fluorescent assays can be difficult to integrate within pharmaceutical production settings. In this paper, we propose a fluorescence-based assay using an optofluidic hollow-core photonic crystal fibre. Exploiting the fibre's waveguide modes to collect fluorescence, the system can quantitatively and specifically measure influenza concentrations below 10particles per ml in under 5 h, using less than 200l of sample. We propose that, with future improvements, this approach could ultimately be integrated directly into production lines to provide continuous feedback on virus concentration, enabling more effective optimisation of process yield.
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5. Quantitative Environmental Monitoring of Pharmaceuticals in Complex Matrices using Isoreticular Ln-BDC Frameworks through Non-Covalent Interactions.
PMID:日期:2026-09-15Lanthanide-based metal-organic frameworks (Ln-MOFs) offer a unique combination of adsorption capabilities and tunable luminescent properties, making them promising materials for environmental applications. This study details the synthesis of a family of isoreticular Ln-MOFs (Ln = Sm, Eu, Tb, Dy) using benzene-1,4-dicarboxylic acid (BDC) as a linker, yielding materials with nanometric crystallite sizes. We explore their efficiency in removing four emerging contaminants (ECs) and systematically evaluate the impact of adsorption on their luminescent properties. While exhibiting a maximum adsorption of 50.2% for naproxen on Dy 2 (BDC) 3 , highlighting the influence of EC functional groups, the materials demonstrate superior functionality as a luminescent sensor. Notably, ciprofloxacin adsorption induces a significant and specific luminescence enhancement (up to 202.24% for Eu-MOF), suggesting great potential for luminescence-based chemical sensing applications. The consistent fitting to the Freundlich model supports the occurrence of adsorption on energetically heterogeneous surfaces. These findings underscore the versatility of Ln-MOF for environmental monitoring through a mechanism that combines adsorption and modulated luminescence.
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6. Evaluating the far-red emitting fluorescence-activating and absorption-shifting tag (frFAST) chemogenetic labeling system for live-cell super-resolution microscopy.
PMID:日期:2026-09-08Fluorogen-activating proteins (FAPs) provide renewable fluorescence signals through continuous fluorogen exchange, resulting in imaging that is virtually resistant to photobleaching, an especially appealing feature for deterministic super-resolution microscopy (SRM) techniques. Furthermore, careful adjustment of the fluorogen concentration may lead to reversible, exchange-driven stochastic blinking, allowing the use of FAPs in stochastic SRM methods. The green-light-excitable, far-red-emitting fluorescence-activating and absorption-shifting tag (frFAST) is particularly well suited for biological imaging due to its favorable spectral properties and live-cell compatibility, however, its performance across different SRM modalities has not been evaluated. Here, we assess frFAST for single-molecule localization microscopy (SMLM), fluctuation-based computational approaches (super-resolution optical fluctuation imaging (SOFI) and extended super-resolution radial fluctuations (eSRRF)), and stimulated emission depletion (STED) microscopy. By tuning fluorogen concentration, we induced reversible stochastic blinking in fixed and live cells, enabling SMLM imaging of cytoskeletal structures and filopodia in mammalian and neuroblastoma cells, without the need for harsh reducing agents. Additionally, we successfully labeled and imaged mitochondrial outer membrane, microtubules, cytoskeleton and histone proteins in live cell STED microscopy. While the renewable nature of the frFAST:HPAR-3OM interaction supports extended live-cell imaging for up to 40 min with limited photobleaching, we found that its performance is strongly modality dependent. In particular, intrinsic photophysical properties limit its suitability for classical SMLM, whereas fluctuation-based methods and live-cell STED microscopy are more compatible with frFAST.
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7. {"_":"A novel rhodamine-based dual-mode fluorogenic probe for sensitive determination of Hg².","sup":[""]}
PMID:日期:2026-09-02Mercury ions (Hg) are highly toxic and persistent heavy metal pollutants that pose serious threats to environmental systems and human health. Therefore, developing sensitive and selective detection strategies for Hgis of great significance. In this study, a rhodamine-based dual-mode fluorescent probe (RB-HG) was developed utilizing dimethylthiocarbamate groups as recognition sites.()()thereby activating both colorimetric and fluorescence dual-channel signals.experiments demonstrate that RB-HG (5.0M) exhibits high selectivity and sensitivity toward Hg. The limits of detection for the fluorescence and colorimetric detection modes are 1.16M and 3.22M, respectively. RB-HG was successfully applied to detect Hgin food samples with excellent recovery rates and to image exogenous Hgin living cells, highlighting its potential for food monitoring and bioimaging applications.
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8. {"_":"A ratiometric fluorescent probe for HSOand its application in environmental and food analysis.","sub":["3"],"sup":["-"]}
PMID:日期:2026-08-26Bisulfite (HSO) is widely used as a preservative and antioxidant in the food industry and chemical manufacturing, yet its excessive residues pose risks to food quality, environmental safety, and human health. To achieve rapid, selective, and quantitative detection of HSO, a ratiometric fluorescent probe based on a benzothiazole-carbazole unit and a benzoindole ion was developed. Upon reaction with HSO, the probe exhibits a distinct dual-emission response, fluorescence intensity decreases at 612 nm and increases at 467 nm, enabling robust ratiometric sensing (/). Owing to its pronounced fluorescence modulation (emission signal change > 145 nm) and high sensitivity (limit of detection = 0.62M), the probe demonstrates reliable performance in aqueous samples and has been successfully integrated into portable test strips and swabs. Furthermore, when coupled with a smartphone-based colorimetric application software, the probe allows convenient and real-time quantification of both gaseous SOand aqueous HSOacross environmental and food-related matrices. This work thus delivers a practical, instrument-free platform for sulfite monitoring in complex real-world settings.
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9. Experimental and simulated FRAP for the quantitative determination of protein diffusion in helical cells.
PMID:日期:2026-08-18Fluorescence recovery after photobleaching (FRAP) is widely used to characterize diffusion in cells, but quantitative interpretation of the data in small prokaryotes requires explicitly accounting for cell geometry. While this has been successfully achieved for spherical and rod-shaped bacteria, analytical approaches developed in these cases are not directly applicable to cells with more complex morphologies. Here, we explore the application of FRAP to helical bacteria using simulations. We show that half-compartment FRAP experiments, where one-half of the cell is photobleached, provide a robust means of characterizing fast protein diffusion. To help with the practical implementation of this technique, we established the relationship between the diffusion coefficient and characteristic fluorescence recovery time as a function of cell length and helical parameters, and for two different ways of estimating the recovery time. As a first application, we report measurements of the diffusion coefficient of the fluorescent protein, mNeonGreen, in the helical bacteriumAMB-1. We find it to be D = 4.9 ± 2.2 μm.sin isosmotic conditions, not significantly different from the value measured in. Although developed for helical bacteria, including spirilla, spirochetes, and vibrios, our framework can readily be extended to cells or compartments with other geometries.
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10. Splitting of luminescence bands of doped quantum dot ensembles in composite with polymers on the silver islands films obtained by chemical deposition method.
PMID:日期:2026-08-18The study demonstrates significant splitting of the impurity luminescence peak of cadmium sulfide quantum dots (QDs) doped with copper and manganese ions in polymer composites on substrates with silver nanoparticles obtained by chemical deposition. The influence of polymer concentration and QD type on the splitting is studied. Our results demonstrate that the probability of splitting occurrence and the spectral separation between the peaks are governed not by the optical density of the film or the size of the silver islands, but by the inter-island distance, highlighting the critical role of spatial arrangement in the collective optical response of the system. It is hypothesized that the observed effect can be caused by coupling between the luminescent nanoparticles, which have long luminescence lifetimes, and the silver nanoislands, which act as nanoresonators.