Methods and Applications in Fluorescence最新文献

Challenges and limitations of molecular resolution fluorescence imaging. 分子分辨率荧光成像的挑战和局限性。

IF 2.4 3区化学

Methods and Applications in Fluorescence Pub Date : 2025-09-17 DOI: 10.1088/2050-6120/ae042b

Dominic A Helmerich, Markus Sauer

{"title":"Challenges and limitations of molecular resolution fluorescence imaging.","authors":"Dominic A Helmerich, Markus Sauer","doi":"10.1088/2050-6120/ae042b","DOIUrl":"10.1088/2050-6120/ae042b","url":null,"abstract":"Super-resolution microscopy (SRM) has revolutionized fluorescence imaging enabling insights into the molecular organization of cells that were previously unconceivable. Latest developments now allow the visualization of individual molecules with nanometer precision and imaging with molecular resolution. However, translating these achievements to imaging under physiological conditions in cells remains challenging. The higher the spatial resolution is pushed by the development of improved SRM methods the more challenging the problems we are confronted when aiming to use them for sub-10 nm fluorescence imaging in cells. It turns out that most developed SRM methods that demonstrate nanometer resolution cannot be directly implemented for molecular resolution imaging in cells. Particularly, fluorescence labeling, i.e. high-density covalent labeling of the molecules of interest with fluorophores with minimal linkage error represents currently a nearly insurmountable obstacle. In addition, even if high labeling densities can be realized it has to be considered that fluorophores can interact via different energy pathways and thus impede super-resolution imaging in the sub-10 nm range. Here, we describe the boundaries, discuss the challenges we must accept and show strategies to circumvent them and achieve true molecular resolution fluorescence imaging under physiological conditions in cells.","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145015789","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An enhanced mountain climbing search algorithm to enable fast and accurate autofocusing in high resolution fluorescence microscopy. 一种增强的爬山搜索算法，使高分辨率荧光显微镜能够快速准确地自动对焦。

IF 2.4 3区化学

Methods and Applications in Fluorescence Pub Date : 2025-09-08 DOI: 10.1088/2050-6120/ae008f

Yuetong Jia, Edward N Ward, Francesca W van Tartwijk, Yutong Yuan, Yuqing Feng, Clemens F Kaminski

{"title":"An enhanced mountain climbing search algorithm to enable fast and accurate autofocusing in high resolution fluorescence microscopy.","authors":"Yuetong Jia, Edward N Ward, Francesca W van Tartwijk, Yutong Yuan, Yuqing Feng, Clemens F Kaminski","doi":"10.1088/2050-6120/ae008f","DOIUrl":"10.1088/2050-6120/ae008f","url":null,"abstract":"Accurate and efficient autofocusing is essential for the automation of fluorescence microscopy, but background noise and shallow depth of field at high magnifications make autofocusing particularly challenging. Here, we present a fast and accurate autofocus algorithm to address these challenges. It is highly effective for high-magnification imaging, while performing equally well for low-magnification imaging tasks. The method is based on the mountain climbing search algorithm and yields improvements on autofocusing precision of up to 200-fold over current methods, whilst offering competitive speed and greatly extended search ranges. Our approach is broadly applicable: it demonstrated good stability and reproducibility across magnifications ranging from 20X to 100X, excels in both live cell imaging and high-resolution fixed sample imaging, and it is compatible with various microscopy techniques without the need for fiducial markers or hardware modifications on existing microscopes. To maximise its accessibility, we constructed a user-friendly interface compatible with the widely used Micromanager software. It generalises well across various imaging modalities and hardware platforms, making it particularly suitable for use in high-resolution screening of candidate drugs.","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144961033","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Advances in ultraviolet microscopy. 紫外显微镜的研究进展。

IF 2.4 3区化学

Methods and Applications in Fluorescence Pub Date : 2025-08-18 DOI: 10.1088/2050-6120/adf800

M McFarlane, G McConnell

引用次数: 0

A blind reconstruction approach for multifocal structured illumination microscopy. 一种多焦结构照明显微镜的盲重建方法。

IF 2.4 3区化学

Methods and Applications in Fluorescence Pub Date : 2025-06-10 DOI: 10.1088/2050-6120/addf12

Haozhen He, Zhiqi Chen, Penghuan Liu

{"title":"A blind reconstruction approach for multifocal structured illumination microscopy.","authors":"Haozhen He, Zhiqi Chen, Penghuan Liu","doi":"10.1088/2050-6120/addf12","DOIUrl":"10.1088/2050-6120/addf12","url":null,"abstract":"Multifocal Structured Illumination Microscopy (MSIM) was initially introduced as a parallel version of image scanning microscopy, aiming to enhance the temporal resolution of the imaging process. Beyond its capacity in super-resolution imaging, MSIM exhibits optical sectioning capabilities akin to confocal microscopy, making it well-suited for imaging thick tissue samples. Traditional MSIM reconstruction algorithms rely on digital pinholes to eliminate out-of-focus signals, demanding precise illumination information. However, controlling and accurately reconstructing illumination patterns can be challenging or impractical in certain experimental settings. To address this, our paper proposes a blind reconstruction method for MSIM that circumvents the need for exact illumination information. Leveraging the stability of the standard deviation for each pixel in illumination, this method achieves optical sectioning effectively and provides approximately 1.76 times better resolution than widefield imaging. The efficacy of our proposed blind reconstruction method for both super-resolution imaging and optical sectioning is validated through both simulations and experimental results.","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144187342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Plasmon-enhanced fluorescence of polyvinylpyrrolidone-embedded CdSeS/ZnS quantum dots on gold film over nanospheres. 纳米球金膜上聚乙烯吡咯烷酮包埋CdSeS/ZnS量子点的等离子体增强荧光。

IF 2.4 3区化学

Methods and Applications in Fluorescence Pub Date : 2025-05-19 DOI: 10.1088/2050-6120/add3c0

A Falamas, C Farcău

{"title":"Plasmon-enhanced fluorescence of polyvinylpyrrolidone-embedded CdSeS/ZnS quantum dots on gold film over nanospheres.","authors":"A Falamas, C Farcău","doi":"10.1088/2050-6120/add3c0","DOIUrl":"10.1088/2050-6120/add3c0","url":null,"abstract":"This study explores the fluorescence enhancement of quantum dots (QDs) by gold film over nanospheres (AuFoN) plasmonic substrates, focusing on how a polymer matrix and plasmon resonances of the substrate affect the fluorescence properties of QDs. It was observed that polyvinylpyrrolidone (PVP) facilitated the uniform distribution of QDs on the surface of the AuFoN by simple drop-coating, avoiding aggregation during solvent evaporation. Progressive fluorescence redshifts and intensity enhancement were observed when moving from QDs on glass substrates to planar Au, and most pronouncedly, to nanostructured AuFoN substrates. The fluorescence enhancement was further analyzed by varying the diameter of the polystyrene spheres used in AuFoN fabrication, revealing that substrates based on 600-700 nm spheres provided the strongest fluorescence amplification due to stronger localized electromagnetic fields. Time-resolved fluorescence measurements revealed two primary fluorescence lifetime components for QDs on AuFoN: a short component linked to non-radiative plasmonic energy transfer and a long component representing intrinsic QDs emission. By optimizing sphere size, Au nanostructured films can be tailored to control QDs fluorescence lifetimes and intensity, advancing their use in biosensing, photonics, and other fluorescence-based technologies. This work enhances our understanding of how substrate design and matrix effects impact QDs fluorescence, providing a pathway for precisely engineered Surface Enhanced Fluorescence (SEF) platforms suited to various applications in optical sensing and more general photonics.","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144001928","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Spectrofluorometric determination of Fe³⁺, Pd²⁺, and Sn²⁺ions using carbon nanodots derived from hazelnut shells. 用榛子壳碳纳米点分光光度法测定Fe3+、Pd2+和Sn2+离子。

IF 2.4 3区化学

Methods and Applications in Fluorescence Pub Date : 2025-05-12 DOI: 10.1088/2050-6120/add1aa

Abidin Gümrükçüoğlu, Miraç Ocak, Sevgi Kolaylı, Saliha Dinç, Ilknur Altin, Serhat Gün, Ümmühan Turgut Ocak

{"title":"Spectrofluorometric determination of Fe3+, Pd2+, and Sn2+ions using carbon nanodots derived from hazelnut shells.","authors":"Abidin Gümrükçüoğlu, Miraç Ocak, Sevgi Kolaylı, Saliha Dinç, Ilknur Altin, Serhat Gün, Ümmühan Turgut Ocak","doi":"10.1088/2050-6120/add1aa","DOIUrl":"https://doi.org/10.1088/2050-6120/add1aa","url":null,"abstract":"Carbon nanodots (CNDs) were synthesized from hazelnut shells using the pyrolysis method in an oven at temperatures ranging from 250 to 400 °C and reaction times between 1 and 3 h. The structural and optical properties of the CNDs, which exhibited strong blue fluorescence under a UV lamp (365 nm), were characterized using UV-vis absorption spectroscopy, fluorescence spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, x-ray diffraction (XRD), x-ray photoelectron spectroscopy (XPS), and transmission electron microscopy (TEM). The quantum yield of the CNDs was determined to be between 2.2% and 7.8%. The effect of various cations on the fluorescence spectra of the CNDs was investigated using fluorescence spectrometry. Among the synthesized CNDs, those prepared via pyrolysis for 3 h at 300 °C, 315 °C, and 350 °C (designated as HS300-3, HS315-3, and HS350-3, respectively) exhibited selective fluorescence quenching in the presence of Fe3+, Sn2+, and Pd2+. These CNDs were applied for the determination of Fe3+in tap water and soil samples, while HS315-3 was also employed for the detection of Sn2+and Pd2+in tap water. The limits of detection (LOD) were 4.5 μg l-1for Fe3+and 15.8 μg l-1for both Sn2+and Pd2+. The accuracy of the methods was validated through spiked recovery experiments with tap water samples and by analyzing a certified reference material (CRM-SA-C Sandy Soil C).","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":"13 3","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144012473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Selective detection of Cr(III) among various chromium oxidation states in aqueous environments. 水环境中不同氧化态Cr（III）的选择性检测。

IF 2.4 3区化学

Methods and Applications in Fluorescence Pub Date : 2025-05-07 DOI: 10.1088/2050-6120/add080

Mengyu Yin, Long Yu, Kaili Wu, Yifei Sun, Yunfei Lu, Zhaojing Yuan, Wenyuan Jiang, Xiangxue Wang, Suhua Wang

{"title":"Selective detection of Cr(III) among various chromium oxidation states in aqueous environments.","authors":"Mengyu Yin, Long Yu, Kaili Wu, Yifei Sun, Yunfei Lu, Zhaojing Yuan, Wenyuan Jiang, Xiangxue Wang, Suhua Wang","doi":"10.1088/2050-6120/add080","DOIUrl":"https://doi.org/10.1088/2050-6120/add080","url":null,"abstract":"Chromium (III), as a significant environmental pollutant, poses a serious threat to human health when accumulated in excess, making it imperative to develop highly sensitive and rapid on-site detection methods. In this study, a Eu3+-based metal-organic framework (MOF) was successfully synthesized using a dual-ligand strategy, enabling ratiometric fluorescence detection of Cr(III). During the detection process, distinct changes in the fluorescence ratio were observed, accompanied by a notable color shift from red to blue, providing a clear visual cue. Aromatic π-conjugated organic ligands were selected to enhance the photoluminescence properties, while a second ligand was introduced to optimize the crystal size and pore structure of the MOF. The experimental results demonstrated that the MOF material exhibited outstanding ratiometric fluorescence properties for Cr(III) detection, with high stability. Additionally, the material showed excellent selectivity, anti-interference ability, and sensitivity for detecting Cr(III) in water environments, with a detection range of 0-24 μM and detection limit as low as 59 nM. Further investigation revealed that the changes in ratiometric fluorescence signals were induced by Cr(III)-specific partial collapse of the crystal structure and ligand release. Finally, a MOF-based test strip was developed, where the vivid fluorescence color changes enabled visual detection by the naked eye. The test strip also demonstrated good recovery efficiency in real water samples, further confirming the material's potential as a real-time smart sensor for Cr(III) detection.","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":"13 3","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144018470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Quantum dots - polymer films as the reversible luminescent wide range temperature sensors. 量子点-聚合物薄膜作为可逆发光宽范围温度传感器。

IF 2.4 3区化学

Methods and Applications in Fluorescence Pub Date : 2025-03-06 DOI: 10.1088/2050-6120/adb9b6

Dmitriy O Sagdeev, Yuriy G Galyametdinov

引用次数: 0

Microwave synthesis of molybdenum disulfide quantum dots and the application in bilirubin sensing. 微波合成二硫化钼量子点及其在胆红素传感中的应用。

IF 2.4 3区化学

Methods and Applications in Fluorescence Pub Date : 2025-01-30 DOI: 10.1088/2050-6120/ada99b

Shu Pang, Xinyu Chang, Lin Xu, Jiazhen Wu

引用次数: 0

Cell viability assessment by using GelRed/SYTO 9-based double staining. 采用GelRed/SYTO - 9双染色法评估细胞活力。