Pub Date : 2024-04-11DOI: 10.1088/2050-6120/ad379f
Eduardo D Wannaz, Rocío Medina, Sheila D Pardo, Gonzalo P Barbero Medina, Andrés Blanco, María Julieta Salazar
This study evaluated the reliability of portable X-ray fluorescence (pXRF) in Pb2+adsorption kinetics and isotherm experiments using soybean straw biochar. The research aimed to compare pXRF results with those obtained through traditional atomic absorption spectrometry (AAS). Soybean straw biochar, produced at 400 °C, was employed as the adsorbent for Pb2+. The efficiency of adsorption was assessed using Langmuir and Freundlich models. The kinetics of Pb2+adsorption was analysed through pseudo-first-order and pseudo-second-order models. The pseudo-second-order model described the kinetics of Pb2+adsorption on biochar better than the pseudo-first order model. Importantly, the pXRF technique demonstrated comparable results to those of AAS, making it a reliable and resource-efficient method for studying Pb2+kinetics. The results of the isotherm analyses fit the Langmuir model, indicating a desirable and irreversible adsorption of Pb2+on biochar. PXRF measurements on biochar allowed simultaneous observations of Pb2+adsorption and K+and Ca2+desorption, highlighting ionic exchange as the primary adsorption mechanism. In conclusion, our results showcased the applicability of pXRF for Pb+2adsorption studies in biochars, offering a valuable alternative to traditional methods. The findings contribute to the understanding of biochar as an effective adsorbent for heavy metals, emphasizing the potential of pXRF for cost-effective and efficient environmental research. In this study, we present a novel and detailed procedure that will allow other researchers to continue their studies on Pb2+adsorption on biochar or similar matrices, significantly reducing the resources and time used and enabling the simultaneous study of the behavior of other ions participating in the process.
{"title":"Measurement of elements by portable x-ray fluorescence spectrometry for the study of adsorption processes: the case of Pb<sup>2+</sup>adsorption on soybean straw biochar.","authors":"Eduardo D Wannaz, Rocío Medina, Sheila D Pardo, Gonzalo P Barbero Medina, Andrés Blanco, María Julieta Salazar","doi":"10.1088/2050-6120/ad379f","DOIUrl":"10.1088/2050-6120/ad379f","url":null,"abstract":"<p><p>This study evaluated the reliability of portable X-ray fluorescence (pXRF) in Pb<sup>2+</sup>adsorption kinetics and isotherm experiments using soybean straw biochar. The research aimed to compare pXRF results with those obtained through traditional atomic absorption spectrometry (AAS). Soybean straw biochar, produced at 400 °C, was employed as the adsorbent for Pb<sup>2+</sup>. The efficiency of adsorption was assessed using Langmuir and Freundlich models. The kinetics of Pb<sup>2+</sup>adsorption was analysed through pseudo-first-order and pseudo-second-order models. The pseudo-second-order model described the kinetics of Pb<sup>2+</sup>adsorption on biochar better than the pseudo-first order model. Importantly, the pXRF technique demonstrated comparable results to those of AAS, making it a reliable and resource-efficient method for studying Pb<sup>2+</sup>kinetics. The results of the isotherm analyses fit the Langmuir model, indicating a desirable and irreversible adsorption of Pb<sup>2+</sup>on biochar. PXRF measurements on biochar allowed simultaneous observations of Pb<sup>2+</sup>adsorption and K<sup>+</sup>and Ca<sup>2+</sup>desorption, highlighting ionic exchange as the primary adsorption mechanism. In conclusion, our results showcased the applicability of pXRF for Pb<sup>+2</sup>adsorption studies in biochars, offering a valuable alternative to traditional methods. The findings contribute to the understanding of biochar as an effective adsorbent for heavy metals, emphasizing the potential of pXRF for cost-effective and efficient environmental research. In this study, we present a novel and detailed procedure that will allow other researchers to continue their studies on Pb<sup>2+</sup>adsorption on biochar or similar matrices, significantly reducing the resources and time used and enabling the simultaneous study of the behavior of other ions participating in the process.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140288542","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}
Pub Date : 2024-04-09DOI: 10.1088/2050-6120/ad3890
Jin-Zhou Liu, Rui-Xian Zhao, Qian-Wei Yin, Hai-Chi Zhang, Rong Sheng Li, Jian Ling, Qiue Cao
Lead halide perovskite nanocrystals (PNCs) have attracted intense attention due to their excellent optoelectronic properties. In this work, a series of water-stable CsPb(Br/I)3PNCs fluorescent probes were prepared using an anion exchange method. It was found that the PNCs probes could be used to detect ascorbic acid (AA) in water, and interestingly, the FL spectra of the PNCs probes can be adjusted by controlling the concentration of KI in anion exchange to improve the detection selectivity of AA. The high sensitivity and selectivity make CsPb(Br/I)3PNCs an ideal material for AA sensing. The concentration of AA can be linearly measured in the range from 0.01 to 50μM, with a detection limit of 4.2 nM. The reason for the enhanced FL of CsPb(Br/I)3PNCs was studied, and it is considered that AA causes the aggregation of CsPb(Br/I)3PNCs. This strategy of improving the selectivity of the probe to the substrate by adjusting the spectrum will significantly expand the application of PNCs in the field of analysis and detection.
卤化铅包晶纳米晶体(PNCs)因其优异的光电特性而备受关注。本研究采用阴离子交换法制备了一系列水稳定性 CsPb(Br/I)3 PNCs 荧光探针。有趣的是,通过控制阴离子交换中 KI 的浓度来调节 PNCs 探针的荧光光谱,可以提高对 AA 的检测选择性。CsPb(Br/I)3 PNCs 的高灵敏度和高选择性特点使其成为 AA 检测的理想材料。研究发现,CsPb(Br/I)3 PNCs FL 增强的原因是 AA 引起了 CsPb(Br/I)3 PNCs 的聚集。这种通过调整光谱来提高探针对底物的选择性的策略将大大拓展 PNCs 在分析和检测领域的应用。
{"title":"Selective detection of ascorbic acid by tuning the composition and fluorescence of the cesium lead halide perovskite nanocrystals.","authors":"Jin-Zhou Liu, Rui-Xian Zhao, Qian-Wei Yin, Hai-Chi Zhang, Rong Sheng Li, Jian Ling, Qiue Cao","doi":"10.1088/2050-6120/ad3890","DOIUrl":"10.1088/2050-6120/ad3890","url":null,"abstract":"<p><p>Lead halide perovskite nanocrystals (PNCs) have attracted intense attention due to their excellent optoelectronic properties. In this work, a series of water-stable CsPb(Br/I)<sub>3</sub>PNCs fluorescent probes were prepared using an anion exchange method. It was found that the PNCs probes could be used to detect ascorbic acid (AA) in water, and interestingly, the FL spectra of the PNCs probes can be adjusted by controlling the concentration of KI in anion exchange to improve the detection selectivity of AA. The high sensitivity and selectivity make CsPb(Br/I)<sub>3</sub>PNCs an ideal material for AA sensing. The concentration of AA can be linearly measured in the range from 0.01 to 50<i>μ</i>M, with a detection limit of 4.2 nM. The reason for the enhanced FL of CsPb(Br/I)<sub>3</sub>PNCs was studied, and it is considered that AA causes the aggregation of CsPb(Br/I)<sub>3</sub>PNCs. This strategy of improving the selectivity of the probe to the substrate by adjusting the spectrum will significantly expand the application of PNCs in the field of analysis and detection.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140306191","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}
Pub Date : 2024-04-05DOI: 10.1088/2050-6120/ad3892
Igor D Romanishkin, Ekaterina V Akhlyustina, Gennady A Meerovich, Victor B Loschenov, Marina G Strakhovskaya
Polycationic photosensitizers (PS) are not susceptible to aggregation in solutions, but their high local concentrations in Gram-negative bacteria can be sufficient for aggregation and reduced effectiveness of antibacterial photodynamic treatment. By measuring fluorescence spectra and kinetics we were able to evaluate the degree of aggregation of polycationic PS ZnPcChol8in Gram-negative bacteria E.coliK12 TG1. Binding of ZnPcChol8toE.coliK12 TG1 leads to an appearance of groups of molecules with shorter PS fluorescence lifetime, a decrease in fluorescence intensity and a shift in the fluorescence spectral maximum. However, we evaluated that about 88% of the fluorescing PS molecules in the bacteria were in an unaggregated state, which indicates only a small reduction in the generation of reactive oxygen species.
{"title":"Оn the aggregation of polycationic photosensitizer upon binding to Gram-negative bacteria.","authors":"Igor D Romanishkin, Ekaterina V Akhlyustina, Gennady A Meerovich, Victor B Loschenov, Marina G Strakhovskaya","doi":"10.1088/2050-6120/ad3892","DOIUrl":"10.1088/2050-6120/ad3892","url":null,"abstract":"<p><p>Polycationic photosensitizers (PS) are not susceptible to aggregation in solutions, but their high local concentrations in Gram-negative bacteria can be sufficient for aggregation and reduced effectiveness of antibacterial photodynamic treatment. By measuring fluorescence spectra and kinetics we were able to evaluate the degree of aggregation of polycationic PS ZnPcChol<sub>8</sub>in Gram-negative bacteria E.<i>coli</i>K12 TG1. Binding of ZnPcChol<sub>8</sub>to<i>E</i>.<i>coli</i>K12 TG1 leads to an appearance of groups of molecules with shorter PS fluorescence lifetime, a decrease in fluorescence intensity and a shift in the fluorescence spectral maximum. However, we evaluated that about 88% of the fluorescing PS molecules in the bacteria were in an unaggregated state, which indicates only a small reduction in the generation of reactive oxygen species.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140306192","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}
Pub Date : 2024-04-05DOI: 10.1088/2050-6120/ad3891
Jiazhen Zhang, Yicong Jia, Xuan Tong, Hangyu Zhou, Le Zhang, Yue Yang, Xu Ji
Food contaminants pose a danger to human health, but rapid, sensitive and reliable food safety detection methods can offer a solution to this problem. In this study, an optical fiber ratiometric fluorescence sensing system based on carbon dots (CDs) and o-phenylenediamine (OPD) was constructed. The ratiometric fluorescence response of Cu2+ and thiram was carried out by the fluorescence resonance energy transfer (FRET) between CDs and 2,3−diaminophenazine (ox-OPD, oxidized state o-phenylenediamine). The oxidation of OPD by Cu2+ resulted in the formation of ox-OPD, which quenched the fluorescence of CDs and exhibited a new emission peak at 573 nm. The formation of a [dithiocarbamate-Cu2+] (DTC-Cu2+) complex by reacting thiram with Cu2+, inhibits the OPD oxidation reaction triggered by Cu2+, thus turning off the fluorescence signal of OPD-Cu2+. The as-established detection system presented excellent sensitivity and selectivity for the detection of Cu2+ and thiram in the ranges of 1 ∼ 100 μM and 5 ∼ 50 μM, respectively. The lowest detection limits were 0.392 μM for Cu2+ and 0.522 μM for thiram. Furthermore, actual sample analysis indicated that the sensor had the potential for Cu2+ and thiram assays in real sample analysis.
{"title":"Portable ratiometric fluorescence detection of Cu2+ and thiram","authors":"Jiazhen Zhang, Yicong Jia, Xuan Tong, Hangyu Zhou, Le Zhang, Yue Yang, Xu Ji","doi":"10.1088/2050-6120/ad3891","DOIUrl":"https://doi.org/10.1088/2050-6120/ad3891","url":null,"abstract":"Food contaminants pose a danger to human health, but rapid, sensitive and reliable food safety detection methods can offer a solution to this problem. In this study, an optical fiber ratiometric fluorescence sensing system based on carbon dots (CDs) and o-phenylenediamine (OPD) was constructed. The ratiometric fluorescence response of Cu<sup>2+</sup> and thiram was carried out by the fluorescence resonance energy transfer (FRET) between CDs and 2,3−diaminophenazine (ox-OPD, oxidized state o-phenylenediamine). The oxidation of OPD by Cu<sup>2+</sup> resulted in the formation of ox-OPD, which quenched the fluorescence of CDs and exhibited a new emission peak at 573 nm. The formation of a [dithiocarbamate-Cu<sup>2+</sup>] (DTC-Cu<sup>2+</sup>) complex by reacting thiram with Cu<sup>2+</sup>, inhibits the OPD oxidation reaction triggered by Cu<sup>2+</sup>, thus turning off the fluorescence signal of OPD-Cu<sup>2+</sup>. The as-established detection system presented excellent sensitivity and selectivity for the detection of Cu<sup>2+</sup> and thiram in the ranges of 1 ∼ 100 <italic toggle=\"yes\">μ</italic>M and 5 ∼ 50 <italic toggle=\"yes\">μ</italic>M, respectively. The lowest detection limits were 0.392 <italic toggle=\"yes\">μ</italic>M for Cu<sup>2+</sup> and 0.522 <italic toggle=\"yes\">μ</italic>M for thiram. Furthermore, actual sample analysis indicated that the sensor had the potential for Cu<sup>2+</sup> and thiram assays in real sample analysis.","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":"57 1","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140610195","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}
Pub Date : 2024-03-26DOI: 10.1088/2050-6120/ad326e
Mahla Shahsavar Gocmen, Ayse Dulda
Lanthanide-doped fluoride nanocrystals (NCs) exhibit excellent optical features, including upconversion and downconversion luminescence (UCL and DCL), that can be utilized in a variety of applications. In this study, we have successfully demonstrated the photoluminescence behavior of triple-doped NaYF4: Yb3+, Er3+, Pr3+NCs in the Vis-NIR region. Herein, highly monodisperse hexagonal phase NaYF4: Yb0.2, Er0.02, Prxnanocrystals in various Pr3+(x = 0, 0.1, 0.5, and 1 mol %) concentration with ∼22 nm diameter synthesized by thermal decomposition technique. The photoluminescence studies for all samples were performed under 980 nm laser excitation. The luminescence intensity of Er3+including blue (407 nm), green (520 and 540 nm), red (654 nm), and near-infrared (845 nm and 1530 nm) emissions was significantly quenched and Pr3+emission intensity at 1290 nm (Pr3+:1G4→3H5) changes irregularly upon doping with Pr3+ions. Furthermore, we performed the excitation power dependence and decay time analysis to investigate the energy transfer and upconversion mechanisms of samples. These findings indicate that the presence of praseodymium strongly reduces emission intensities due to abundant cross-relaxation channels. In addition, particle size is an efficient factor, shedding light on the influence of Pr3+on the energy transfer and upconversion mechanisms of the fluorides.
{"title":"Effect of Pr<sup>3+</sup>concentration in luminescence properties & upconversion mechanism of triple doped NaYF<sub>4</sub>: Yb<sup>3+</sup>, Er<sup>3+</sup>, Pr<sup>3</sup>.","authors":"Mahla Shahsavar Gocmen, Ayse Dulda","doi":"10.1088/2050-6120/ad326e","DOIUrl":"10.1088/2050-6120/ad326e","url":null,"abstract":"<p><p>Lanthanide-doped fluoride nanocrystals (NCs) exhibit excellent optical features, including upconversion and downconversion luminescence (UCL and DCL), that can be utilized in a variety of applications. In this study, we have successfully demonstrated the photoluminescence behavior of triple-doped NaYF<sub>4</sub>: Yb<sup>3+</sup>, Er<sup>3+</sup>, Pr<sup>3+</sup>NCs in the Vis-NIR region. Herein, highly monodisperse hexagonal phase NaYF<sub>4</sub>: Yb<sub>0.2</sub>, Er<sub>0.02</sub>, Pr<sub>x</sub>nanocrystals in various Pr<sup>3+</sup>(x = 0, 0.1, 0.5, and 1 mol %) concentration with ∼22 nm diameter synthesized by thermal decomposition technique. The photoluminescence studies for all samples were performed under 980 nm laser excitation. The luminescence intensity of Er<sup>3+</sup>including blue (407 nm), green (520 and 540 nm), red (654 nm), and near-infrared (845 nm and 1530 nm) emissions was significantly quenched and Pr<sup>3+</sup>emission intensity at 1290 nm (Pr<sup>3+</sup>:<sup>1</sup>G<sub>4</sub>→<sup>3</sup>H<sub>5</sub>) changes irregularly upon doping with Pr<sup>3+</sup>ions. Furthermore, we performed the excitation power dependence and decay time analysis to investigate the energy transfer and upconversion mechanisms of samples. These findings indicate that the presence of praseodymium strongly reduces emission intensities due to abundant cross-relaxation channels. In addition, particle size is an efficient factor, shedding light on the influence of Pr<sup>3+</sup>on the energy transfer and upconversion mechanisms of the fluorides.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140102005","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}
Pub Date : 2024-03-21DOI: 10.1088/2050-6120/ad326d
Matías E Carranza, Hugo M Elero, Pedro J Peña Hernández, Alicia V Veglia
Absorption and fluorescence spectra of the nitrogen polycyclic aromatic hydrocarbon carbazole (CZL) were analyzed with native cyclodextrins (CD;α,β,γ); derivatizedCD(hydroxypropyl-β-cyclodextrin,HPCD; methyl-β-cyclodextrin,MeCD) and p-sulfonated calixarenes (SCAn, with n = 6 and 8) macrocycles. The results showed a slight increase in the absorbance ofCZLwithCD, but the mixture ofCZLwithSCAshowed lower absorption than the sum of the individual spectra. Also, changes in fluorescence were observed by adding the macrocycles, quenching withSCA, and significant increases withCD. The higher fluorescence enhancement was withHPCDrationalized as a complex formation with 1:1 stoichiometry, with an average value for the association constant (KA) of (12 ± 1) x 102M-1, and a quantum yield ratio between the complexedCZLand freeCZL(ΦCZL-HPCD/ΦCZL) of (1.56 ± 0.02) at neutral pH and 25.0 °C. These increases in fluorescence were used as an on-fluorescence switch to develop a supramolecular analytical method forCZLin aqueous samples. The best analytical parameters were inHPCD(LOD = 1.41 ± 0.01 ng mL-1). The method was validated in aqueous samples of river and tap water with recoveries between 96%-104%. The proposed supramolecular method is quick, direct, selective and represents an alternative and low-cost analysis method.
{"title":"Calixarenes and cyclodextrins as off- and on-fluorescence probes for carbazole.","authors":"Matías E Carranza, Hugo M Elero, Pedro J Peña Hernández, Alicia V Veglia","doi":"10.1088/2050-6120/ad326d","DOIUrl":"10.1088/2050-6120/ad326d","url":null,"abstract":"<p><p>Absorption and fluorescence spectra of the nitrogen polycyclic aromatic hydrocarbon carbazole (<b>CZL</b>) were analyzed with native cyclodextrins (<b>CD</b>;<i>α</i>,<i>β</i>,<i>γ</i>); derivatized<b>CD</b>(hydroxypropyl-<i>β</i>-cyclodextrin,<b>HPCD</b>; methyl-<i>β</i>-cyclodextrin,<b>MeCD</b>) and p-sulfonated calixarenes (<b>SCAn</b>, with n = 6 and 8) macrocycles. The results showed a slight increase in the absorbance of<b>CZL</b>with<b>CD</b>, but the mixture of<b>CZL</b>with<b>SCA</b>showed lower absorption than the sum of the individual spectra. Also, changes in fluorescence were observed by adding the macrocycles, quenching with<b>SCA</b>, and significant increases with<b>CD</b>. The higher fluorescence enhancement was with<b>HPCD</b>rationalized as a complex formation with 1:1 stoichiometry, with an average value for the association constant (<i>K</i><sub><i>A</i></sub>) of (12 ± 1) x 10<sup>2</sup>M<sup>-1</sup>, and a quantum yield ratio between the complexed<b>CZL</b>and free<b>CZL</b>(Φ<sup>CZL-HPCD</sup>/Φ<sup>CZL</sup>) of (1.56 ± 0.02) at neutral pH and 25.0 °C. These increases in fluorescence were used as an on-fluorescence switch to develop a supramolecular analytical method for<b>CZL</b>in aqueous samples. The best analytical parameters were in<b>HPCD</b>(LOD = 1.41 ± 0.01 ng mL<sup>-1</sup>). The method was validated in aqueous samples of river and tap water with recoveries between 96%-104%. The proposed supramolecular method is quick, direct, selective and represents an alternative and low-cost analysis method.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140102004","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}
Pub Date : 2024-03-19DOI: 10.1088/2050-6120/ad31b8
Micaela Toscani, Axel M Lacapmesure
Here we apply the SUPPOSe algorithm on images acquired using Stimulated Emission Depletion (STED) microscopy with the aim of improving the resolution limit achieved. We processed images of the nuclear pore complex (NPC) from cell lines in which the Nup96 nucleoporin was endogenously labeled. This reference protein forms a ring whose diameter is ∼107 nm with 8 corners ∼42 nm apart from each other. The stereotypic arrangement of proteins in the NPC has been used as reference structures to characterize the performance of a variety of microscopy techniques. STED microscopy images resolve the ring arrangement but not the eightfold symmetry of the NPC. After applying the SUPPOSe algorithm to the STED images, we were able to solve the octagonal structure of the NPC. After processing 562 single NPC, the average radius of the NPC was found to beR= 54.2 ± 2.9 nm, being consistent with the theoretical distances of this structure. To verify that the solutions obtained are compatible with a NPC-type geometry, we rotate the solutions to optimally fit an eightfold-symmetric pattern and we count the number of corners that contain at least one localization. Fitting a probabilistic model to the histogram of the number of bright corners gives an effective labeling efficiency of 31%, which is in agreement with the values reported in for other cell lines and ligands used in Single Molecule Localization microscopy, showing that SUPPOSe can reliably retrieve sub-resolution, nanoscale objects from single acquisitions even in noisy conditions.
{"title":"Improving STED microscopy with SUPPOSe: enhancing resolution from a single-image.","authors":"Micaela Toscani, Axel M Lacapmesure","doi":"10.1088/2050-6120/ad31b8","DOIUrl":"10.1088/2050-6120/ad31b8","url":null,"abstract":"<p><p>Here we apply the SUPPOSe algorithm on images acquired using Stimulated Emission Depletion (STED) microscopy with the aim of improving the resolution limit achieved. We processed images of the nuclear pore complex (NPC) from cell lines in which the Nup96 nucleoporin was endogenously labeled. This reference protein forms a ring whose diameter is ∼107 nm with 8 corners ∼42 nm apart from each other. The stereotypic arrangement of proteins in the NPC has been used as reference structures to characterize the performance of a variety of microscopy techniques. STED microscopy images resolve the ring arrangement but not the eightfold symmetry of the NPC. After applying the SUPPOSe algorithm to the STED images, we were able to solve the octagonal structure of the NPC. After processing 562 single NPC, the average radius of the NPC was found to be<i>R</i>= 54.2 ± 2.9 nm, being consistent with the theoretical distances of this structure. To verify that the solutions obtained are compatible with a NPC-type geometry, we rotate the solutions to optimally fit an eightfold-symmetric pattern and we count the number of corners that contain at least one localization. Fitting a probabilistic model to the histogram of the number of bright corners gives an effective labeling efficiency of 31%, which is in agreement with the values reported in for other cell lines and ligands used in Single Molecule Localization microscopy, showing that SUPPOSe can reliably retrieve sub-resolution, nanoscale objects from single acquisitions even in noisy conditions.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140065499","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}
Pub Date : 2024-03-15DOI: 10.1088/2050-6120/ad2f39
Alain Bolaño Alvarez, Steffen B Petersen
We here report the formation of a turbid-gel phase in acrylic cuvettes upon exposure to pure Dimethyl Sulfoxide (DMSO) at room temperature. The observed phenomenon occurred over a 10 h to 14 h incubation in the presence of environmental oxygen. After the turbid gel was removed from the cuvette, it became a white solid exhibiting unique emission behavior. The formation of the turbid-gel phase was accelerated upon exposure to UV 295 LED pulses of light from 6 h to 8 h. Surprisingly, subsequent exposure of the white solid to a few microliters of pure DMSO and vortexing resulted in its transformation into a transparent gel state in just a few minutes, eventually acquiring transparent and liquid properties. Additionally, the white-solid phase can load other molecules, such as Resveratrol and Quercetin, leading to shifts in the respective emission spectra compared with the same molecule in liquid and pure DMSO. These novel findings highlight the interaction between UV photons, oxygen, DMSO and Acrylic, and potentially distort fluorescence spectroscopy experiments.
{"title":"Formation of gel and solid phases in acrylic cuvettes upon exposure to DMSO, oxygen and light: implications for fluorescence spectroscopy.","authors":"Alain Bolaño Alvarez, Steffen B Petersen","doi":"10.1088/2050-6120/ad2f39","DOIUrl":"10.1088/2050-6120/ad2f39","url":null,"abstract":"<p><p>We here report the formation of a turbid-gel phase in acrylic cuvettes upon exposure to pure Dimethyl Sulfoxide (DMSO) at room temperature. The observed phenomenon occurred over a 10 h to 14 h incubation in the presence of environmental oxygen. After the turbid gel was removed from the cuvette, it became a white solid exhibiting unique emission behavior. The formation of the turbid-gel phase was accelerated upon exposure to UV 295 LED pulses of light from 6 h to 8 h. Surprisingly, subsequent exposure of the white solid to a few microliters of pure DMSO and vortexing resulted in its transformation into a transparent gel state in just a few minutes, eventually acquiring transparent and liquid properties. Additionally, the white-solid phase can load other molecules, such as Resveratrol and Quercetin, leading to shifts in the respective emission spectra compared with the same molecule in liquid and pure DMSO. These novel findings highlight the interaction between UV photons, oxygen, DMSO and Acrylic, and potentially distort fluorescence spectroscopy experiments.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140012915","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}
Pub Date : 2024-03-12DOI: 10.1088/2050-6120/ad2f3a
Conghuan Lin, Huizhi Du
Forsythoside E is one secondary metabolite ofForsythia suspensa(Thunb.) Vahl. In the study, the interactions between forsythoside E and two types of cholinesterases, acetylcholinesterase and butyrylcholinesterase were investigated in the different conditions. Forsythoside E increased the fluorescence intensity of acetylcholinesterase but quenched the fluorescence of butyrylcholinesterase. Aβ25-35used in the study may not form complexes with cholinesterases, and did not affect the interaction between forsythoside E and cholinesterases. The charged quaternary group of AsCh interacted with the 'anionic' subsite in acetylcholinesterase, which did not affect the interaction between forsythoside E and acetylcholinesterase. The enhancement rate of forsythoside E to acetylcholinesterase fluorescence from high to low was acid solution (pH 6.4), neutral solution (pH 7.4) and alkaline solution (pH 8.0), while the reduction rate of forsythoside E to butyrylcholinesterase fluorescence was in reverse order. Metal ions may interact with cholinesterases, and increased the effects of forsythoside E to cholinesterases fluorescence, in order that Fe3+was the highest, followed by Cu2+, and Mg2+. A forsythoside E-butyrylcholinesterase complex at stoichiometric ratio of 1:1 was spontaneously formed, and the static quenching was the main quenching mode in the process of forsythoside E binding with butyrylcholinesterase. TheKvalues of two complexes were pretty much the same, suggesting that the interaction between cholinesterases and forsythoside E was almost unaffected by acid-base environment and metal ions. Thennumbers of two cholinesterases approximately equaled to one, indicating that there was only one site on each cholinesterase applicable for forsythoside E to bind to.
连翘苷 E 是连翘的一种次生代谢产物。本研究考察了连翘苷 E 与两种胆碱酯酶(乙酰胆碱酯酶和丁酰胆碱酯酶)在不同条件下的相互作用。连翘苷 E 增加了乙酰胆碱酯酶的荧光强度,但淬灭了丁酰胆碱酯酶的荧光。研究中使用的 Aβ25-35 可能不会与胆碱酯酶形成复合物,也不会影响连翘苷 E 与胆碱酯酶之间的相互作用。AsCh 带电荷的季基与乙酰胆碱酯酶中的 "阴离子 "位点相互作用,这并不影响连翘苷 E 与乙酰胆碱酯酶之间的相互作用。连翘苷 E 对乙酰胆碱酯酶荧光的增强率从高到低依次为酸性溶液(pH 6.4)、中性溶液(pH 7.4)和碱性溶液(pH 8.0),而连翘苷 E 对丁酰胆碱酯酶荧光的减弱率则相反。金属离子可能与胆碱酯酶发生作用,增加了连翘苷 E 对胆碱酯酶荧光的影响,依次是 Fe3+ 的影响最大,其次是 Cu2+ 和 Mg2+。在连翘苷 E 与丁酰胆碱酯酶结合的过程中,自发形成了比例为 1:1 的连翘苷 E-丁酰胆碱酯酶复合物,静态淬灭是主要的淬灭方式。两种复合物的 K 值基本相同,表明胆碱酯酶与连翘苷 E 的相互作用几乎不受酸碱环境和金属离子的影响。两种胆碱酯酶的 n 数近似等于 1,表明每种胆碱酯酶上只有一个位点可与连翘苷 E 结合。
{"title":"Interactions between forsythoside E and two cholinesterases at the different conditions: fluorescence sections.","authors":"Conghuan Lin, Huizhi Du","doi":"10.1088/2050-6120/ad2f3a","DOIUrl":"10.1088/2050-6120/ad2f3a","url":null,"abstract":"<p><p>Forsythoside E is one secondary metabolite of<i>Forsythia suspensa</i>(Thunb.) Vahl. In the study, the interactions between forsythoside E and two types of cholinesterases, acetylcholinesterase and butyrylcholinesterase were investigated in the different conditions. Forsythoside E increased the fluorescence intensity of acetylcholinesterase but quenched the fluorescence of butyrylcholinesterase. A<i>β</i><sub>25-35</sub>used in the study may not form complexes with cholinesterases, and did not affect the interaction between forsythoside E and cholinesterases. The charged quaternary group of AsCh interacted with the 'anionic' subsite in acetylcholinesterase, which did not affect the interaction between forsythoside E and acetylcholinesterase. The enhancement rate of forsythoside E to acetylcholinesterase fluorescence from high to low was acid solution (pH 6.4), neutral solution (pH 7.4) and alkaline solution (pH 8.0), while the reduction rate of forsythoside E to butyrylcholinesterase fluorescence was in reverse order. Metal ions may interact with cholinesterases, and increased the effects of forsythoside E to cholinesterases fluorescence, in order that Fe<sup>3+</sup>was the highest, followed by Cu<sup>2+</sup>, and Mg<sup>2+</sup>. A forsythoside E-butyrylcholinesterase complex at stoichiometric ratio of 1:1 was spontaneously formed, and the static quenching was the main quenching mode in the process of forsythoside E binding with butyrylcholinesterase. The<i>K</i>values of two complexes were pretty much the same, suggesting that the interaction between cholinesterases and forsythoside E was almost unaffected by acid-base environment and metal ions. The<i>n</i>numbers of two cholinesterases approximately equaled to one, indicating that there was only one site on each cholinesterase applicable for forsythoside E to bind to.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140012916","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}
Pub Date : 2024-02-08DOI: 10.1088/2050-6120/ad12f7
Dorian Gouzou, Ali Taimori, Tarek Haloubi, Neil Finlayson, Qiang Wang, James R Hopgood, Marta Vallejo
Many medical imaging modalities have benefited from recent advances in Machine Learning (ML), specifically in deep learning, such as neural networks. Computers can be trained to investigate and enhance medical imaging methods without using valuable human resources. In recent years, Fluorescence Lifetime Imaging (FLIm) has received increasing attention from the ML community. FLIm goes beyond conventional spectral imaging, providing additional lifetime information, and could lead to optical histopathology supporting real-time diagnostics. However, most current studies do not use the full potential of machine/deep learning models. As a developing image modality, FLIm data are not easily obtainable, which, coupled with an absence of standardisation, is pushing back the research to develop models which could advance automated diagnosis and help promote FLIm. In this paper, we describe recent developments that improve FLIm image quality, specifically time-domain systems, and we summarise sensing, signal-to-noise analysis and the advances in registration and low-level tracking. We review the two main applications of ML for FLIm: lifetime estimation and image analysis through classification and segmentation. We suggest a course of action to improve the quality of ML studies applied to FLIm. Our final goal is to promote FLIm and attract more ML practitioners to explore the potential of lifetime imaging.
{"title":"Applications of machine learning in time-domain fluorescence lifetime imaging: a review.","authors":"Dorian Gouzou, Ali Taimori, Tarek Haloubi, Neil Finlayson, Qiang Wang, James R Hopgood, Marta Vallejo","doi":"10.1088/2050-6120/ad12f7","DOIUrl":"10.1088/2050-6120/ad12f7","url":null,"abstract":"<p><p>Many medical imaging modalities have benefited from recent advances in Machine Learning (ML), specifically in deep learning, such as neural networks. Computers can be trained to investigate and enhance medical imaging methods without using valuable human resources. In recent years, Fluorescence Lifetime Imaging (FLIm) has received increasing attention from the ML community. FLIm goes beyond conventional spectral imaging, providing additional lifetime information, and could lead to optical histopathology supporting real-time diagnostics. However, most current studies do not use the full potential of machine/deep learning models. As a developing image modality, FLIm data are not easily obtainable, which, coupled with an absence of standardisation, is pushing back the research to develop models which could advance automated diagnosis and help promote FLIm. In this paper, we describe recent developments that improve FLIm image quality, specifically time-domain systems, and we summarise sensing, signal-to-noise analysis and the advances in registration and low-level tracking. We review the two main applications of ML for FLIm: lifetime estimation and image analysis through classification and segmentation. We suggest a course of action to improve the quality of ML studies applied to FLIm. Our final goal is to promote FLIm and attract more ML practitioners to explore the potential of lifetime imaging.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10851337/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138498873","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}