Caenorhabditis elegans (C. elegans) is a typical model organism that has predominantly relied on growth using a bacterial diet, presenting limitations for automated experimentations (mainly due to the requirement of periodic transfer to new plates), and accuracy of results (because of possible interference by bacterial metabolism in liquid media), which makes flexible manipulation and long-term tracking difficult. C. elegans Maintenance Medium (CeMM), a chemically defined sterile liquid medium, holds the potential to solve these problems. Population-level studies of C. elegans cultured in CeMM have shown that nematode development slows, fecundity declines, lifespan increases, lipid and protein stores decrease, and gene expression changes relative to that on a bacterial diet. However, automated cultivation of C. elegans in CeMM, long-term tracking and phenotyping at single-worm level remains challenging. Here, we developed a chamber-array chip (WormChip-1.8) and an integrated and enclosed microfluidic device, CeMM-WormTracker, with the capability of automating fluid control and worm manipulation for single-worm level tracking and phenotyping of C. elegans grown in CeMM. By using a microscope, the CeMM-WormTracker allows for observation of C. elegans development, motility, reproduction, and survival for a long period of time. Our data demonstrate that it is possible to longitudinally track and phenotype the growth and development of nematodes at single-worm level for at least 75 days within the device, enabling comprehensive monitoring of the whole reproductive period and lifespan in sterile liquid culture, which is difficult for well plate-based experiments. Comparing with the results from 96-well plates, the development, activity, and reproduction of nematodes in the microfluidic device seems more stable. Thus, the CeMM-WormTracker provides feasible solutions for automated and high-throughput experimentations in studies of C. elegans, both on the ground and in orbit.
{"title":"CeMM-WormTracker: Long-term, single-worm level tracking and phenotyping of C. elegans cultivated in CeMM on an integrated and enclosed microfluidic device","authors":"Qianqian Yang, Runtao Zhong, Mengyu Wang, Wenbo Chang, Kexin Chen, Yeqing Sun","doi":"10.1039/d5an00394f","DOIUrl":"https://doi.org/10.1039/d5an00394f","url":null,"abstract":"Caenorhabditis elegans (C. elegans) is a typical model organism that has predominantly relied on growth using a bacterial diet, presenting limitations for automated experimentations (mainly due to the requirement of periodic transfer to new plates), and accuracy of results (because of possible interference by bacterial metabolism in liquid media), which makes flexible manipulation and long-term tracking difficult. C. elegans Maintenance Medium (CeMM), a chemically defined sterile liquid medium, holds the potential to solve these problems. Population-level studies of C. elegans cultured in CeMM have shown that nematode development slows, fecundity declines, lifespan increases, lipid and protein stores decrease, and gene expression changes relative to that on a bacterial diet. However, automated cultivation of C. elegans in CeMM, long-term tracking and phenotyping at single-worm level remains challenging. Here, we developed a chamber-array chip (WormChip-1.8) and an integrated and enclosed microfluidic device, CeMM-WormTracker, with the capability of automating fluid control and worm manipulation for single-worm level tracking and phenotyping of C. elegans grown in CeMM. By using a microscope, the CeMM-WormTracker allows for observation of C. elegans development, motility, reproduction, and survival for a long period of time. Our data demonstrate that it is possible to longitudinally track and phenotype the growth and development of nematodes at single-worm level for at least 75 days within the device, enabling comprehensive monitoring of the whole reproductive period and lifespan in sterile liquid culture, which is difficult for well plate-based experiments. Comparing with the results from 96-well plates, the development, activity, and reproduction of nematodes in the microfluidic device seems more stable. Thus, the CeMM-WormTracker provides feasible solutions for automated and high-throughput experimentations in studies of C. elegans, both on the ground and in orbit.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"22 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144603752","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}
Arhama Arhama Ansari, Ayush Ransingh, Soumyo Mukherji, Andrew S Hursthouse, Fiona Henriquez, John Connolly, Suparna Mukherji
The widespread detection of antibiotics in aquatic environments, particularly in effluent-receiving surface waters, poses significant ecological and public health concerns due to their role in promoting antimicrobial resistance. Accurate trace-level antibiotic measurement is essential for environmental risk assessment and improving wastewater treatment strategies. This study presents the development, optimization, and validation of two complementary liquid chromatography-mass spectrometry (LC-MS) workflows for the simultaneous quantification of nine antibiotics across five therapeutic classes in creek water impacted by a Common Effluent Treatment Plant (CETP). The performance of a triple quadrupole LC-MS/MS system (LC-QqQ-MS) was compared to that of a high-resolution Orbitrap mass spectrometer (LC-Orbitrap-HRMS). Both instruments demonstrated excellent linearity (R² > 0.99) and satisfactory recoveries (70–90%) across a wide concentration range. The method detection limits ranged from 0.11 to 0.23 ng L⁻¹ for LC-QqQ-MS and from 0.02 to 0.13 ng L⁻¹ for LC-Orbitrap-HRMS, confirming the superior sensitivity of the high-resolution system approach. Application to real-world creek water samples revealed the ubiquitous presence of multiple antibiotics, with azithromycin and enrofloxacin dominating the detected concentrations, particularly near the CETP discharge point and a nearby waste dumping site. A three-way ANOVA confirmed that antibiotic concentrations were significantly affected by instrument type, sampling site, and antibiotic class along with their interactions. Additionally, non-target screening performed using LC-Orbitrap-HRMS enabled the detection of additional antibiotics belonging to quinolones, sulfonamides and aminoglycosides, further demonstrating the broader analytical scope of high-resolution mass spectrometry. The study highlights the necessity of using advanced analytical tools for the accurate quantification of antibiotics in complex matrices and underscores the environmental risks posed by pharmaceutical pollution in industrial discharge-impacted water bodies.
在水生环境中，特别是在接收废水的地表水中广泛检测到抗生素，由于它们在促进抗菌素耐药性方面的作用，引起了重大的生态和公共卫生问题。准确的痕量抗生素测量对于环境风险评估和改善废水处理策略至关重要。本研究提出了两种互补的液相色谱-质谱（LC-MS）工作流程的开发、优化和验证，用于同时定量受公共污水处理厂（CETP）影响的溪水中五种治疗类别的九种抗生素。将三重四极杆LC-MS/MS系统（LC-QqQ-MS）与高分辨率Orbitrap质谱计（LC-Orbitrap-HRMS）的性能进行了比较。两种仪器都表现出良好的线性度(R²>；0.99)，在较宽的浓度范围内回收率为70-90%。LC-QqQ-MS的检测限为0.11 ~ 0.23 ng L -⁻¹，LC-Orbitrap-HRMS的检测限为0.02 ~ 0.13 ng L -⁻¹，证实了高分辨率系统方法的高灵敏度。应用于实际的溪水样本显示，多种抗生素普遍存在，以阿奇霉素和恩诺沙星的检测浓度占主导地位，特别是在CETP排放点和附近的废物倾倒场附近。三因素方差分析证实，抗生素浓度受仪器类型、采样地点、抗生素类别及其相互作用的显著影响。此外，使用LC-Orbitrap-HRMS进行非靶标筛选，可以检测到喹诺酮类抗生素、磺胺类抗生素和氨基糖苷类抗生素，进一步证明了高分辨率质谱法更广泛的分析范围。该研究强调了使用先进的分析工具对复杂基质中抗生素进行精确定量的必要性，并强调了工业排放影响水体中药物污染所带来的环境风险。
{"title":"Comparative Performance Evaluation of Triple Quadrupole Tandem Mass Spectrometry and Orbitrap High-Resolution Mass Spectrometry for Analysis of Antibiotics in Creek Water Impacted by CETP Discharge","authors":"Arhama Arhama Ansari, Ayush Ransingh, Soumyo Mukherji, Andrew S Hursthouse, Fiona Henriquez, John Connolly, Suparna Mukherji","doi":"10.1039/d5an00482a","DOIUrl":"https://doi.org/10.1039/d5an00482a","url":null,"abstract":"The widespread detection of antibiotics in aquatic environments, particularly in effluent-receiving surface waters, poses significant ecological and public health concerns due to their role in promoting antimicrobial resistance. Accurate trace-level antibiotic measurement is essential for environmental risk assessment and improving wastewater treatment strategies. This study presents the development, optimization, and validation of two complementary liquid chromatography-mass spectrometry (LC-MS) workflows for the simultaneous quantification of nine antibiotics across five therapeutic classes in creek water impacted by a Common Effluent Treatment Plant (CETP). The performance of a triple quadrupole LC-MS/MS system (LC-QqQ-MS) was compared to that of a high-resolution Orbitrap mass spectrometer (LC-Orbitrap-HRMS). Both instruments demonstrated excellent linearity (R² > 0.99) and satisfactory recoveries (70–90%) across a wide concentration range. The method detection limits ranged from 0.11 to 0.23 ng L⁻¹ for LC-QqQ-MS and from 0.02 to 0.13 ng L⁻¹ for LC-Orbitrap-HRMS, confirming the superior sensitivity of the high-resolution system approach. Application to real-world creek water samples revealed the ubiquitous presence of multiple antibiotics, with azithromycin and enrofloxacin dominating the detected concentrations, particularly near the CETP discharge point and a nearby waste dumping site. A three-way ANOVA confirmed that antibiotic concentrations were significantly affected by instrument type, sampling site, and antibiotic class along with their interactions. Additionally, non-target screening performed using LC-Orbitrap-HRMS enabled the detection of additional antibiotics belonging to quinolones, sulfonamides and aminoglycosides, further demonstrating the broader analytical scope of high-resolution mass spectrometry. The study highlights the necessity of using advanced analytical tools for the accurate quantification of antibiotics in complex matrices and underscores the environmental risks posed by pharmaceutical pollution in industrial discharge-impacted water bodies.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"37 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144594835","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}
Alaleh Vaghef-Koodehi,Victor H Perez-Gonzalez,Blanca H Lapizco-Encinas
Insulator based electrokinetic (iEK) devices have emerged as powerful tools for analyzing both nano- and microparticles due to their simplicity, robustness, and ability to integrate linear and nonlinear electrokinetic (EK) effects into a single platform. Recent studies emphasize the importance of nonlinear electrophoresis (EPNL) in particle analysis, for performing separations based on size, shape, and charge differences. Despite these advancements, the development of an empirical equation for predicting particle retention times in iEK-based systems that incorporates EPNL remains limited. This study presents a method for predicting particle retention time in iEK systems in scenarios where the linear EK regime allows for particles migration, while also incorporating EPNL and accounting for particle characteristics, applied electric fields, and microdevice features. Experiments were conducted using eight reference microparticles, grouped into four pairs with similar sizes (3.6 μm to 11.7 μm) but distinct zeta potentials (∼-20 mV and ∼-30 mV), across three distinct iEK microdevices: one with asymmetrical oval-diamond posts, one with symmetrical oval posts, and one postless design. Experimental retention times (tR,e) were measured at applied voltages ranging from 400 V to 1450 V. Using the collected tR,e data, three empirical equations were developed to describe particle velocity, incorporating both linear and nonlinear velocities. Validation with two control particles demonstrated prediction errors below 24% in all devices. These findings underscore the potential of the empirical equations in predicting particle behavior in iEK systems.
{"title":"Predicting the retention time of microparticles in electrokinetic migration.","authors":"Alaleh Vaghef-Koodehi,Victor H Perez-Gonzalez,Blanca H Lapizco-Encinas","doi":"10.1039/d5an00515a","DOIUrl":"https://doi.org/10.1039/d5an00515a","url":null,"abstract":"Insulator based electrokinetic (iEK) devices have emerged as powerful tools for analyzing both nano- and microparticles due to their simplicity, robustness, and ability to integrate linear and nonlinear electrokinetic (EK) effects into a single platform. Recent studies emphasize the importance of nonlinear electrophoresis (EPNL) in particle analysis, for performing separations based on size, shape, and charge differences. Despite these advancements, the development of an empirical equation for predicting particle retention times in iEK-based systems that incorporates EPNL remains limited. This study presents a method for predicting particle retention time in iEK systems in scenarios where the linear EK regime allows for particles migration, while also incorporating EPNL and accounting for particle characteristics, applied electric fields, and microdevice features. Experiments were conducted using eight reference microparticles, grouped into four pairs with similar sizes (3.6 μm to 11.7 μm) but distinct zeta potentials (∼-20 mV and ∼-30 mV), across three distinct iEK microdevices: one with asymmetrical oval-diamond posts, one with symmetrical oval posts, and one postless design. Experimental retention times (tR,e) were measured at applied voltages ranging from 400 V to 1450 V. Using the collected tR,e data, three empirical equations were developed to describe particle velocity, incorporating both linear and nonlinear velocities. Validation with two control particles demonstrated prediction errors below 24% in all devices. These findings underscore the potential of the empirical equations in predicting particle behavior in iEK systems.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"92 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144594215","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}
Continuous glucose level monitoring is essential for the effective treatment of diabetes and overall metabolic health, requiring advanced sensing technologies that provide precise, reliable, and sustained performance in physiological situations. This study presents an integrated enzymatic dual-signal self-powered flexible sensor based on ferrocene-mediated glucose biofuel cells (BFCs), allowing rapid glucose detection through dual signal transduction of current and electrochromic response. The anode (flexible Au/CNT-rGO paper films), functionalized with glucose oxidase/ferrocene (GOD/Fc), catalyzes glucose oxidation to gluconate and generates electrons and outputs the current signal, where Fc facilitates electron transport to mitigate oxygen dependency. The cathode (ITO/PET) provides a visual signal with chitosan@prussian (CS@PB) nano-composite integrated as an electrochromic region. The sensor exhibits a low detection limit of 0.018 mM and a linear detection range from 0.1 to 80 mM, with excellent selectivity and stability. In addition, clinical serum glucose and urine testing experiments validated the sensor's efficacy, demonstrating its potential for biomedical research and clinical applications. The sensor's self-power generation and dual-signal readout provide a promising platform for the development of point-of-care (POC) devices tailored for personalized metabolic health assessment.
{"title":"An integrated dual-signal self-powered flexible sensor based on ferrocene-mediated biofuel cell for glucose detection.","authors":"Zheng Wang,Maruf Ahmed,Jiayuan Zhu,Ying-Zhuo Shen,Meijuan Zhao,Wei Liu,Xiao-Ya Hu,Qin Xu","doi":"10.1039/d5an00633c","DOIUrl":"https://doi.org/10.1039/d5an00633c","url":null,"abstract":"Continuous glucose level monitoring is essential for the effective treatment of diabetes and overall metabolic health, requiring advanced sensing technologies that provide precise, reliable, and sustained performance in physiological situations. This study presents an integrated enzymatic dual-signal self-powered flexible sensor based on ferrocene-mediated glucose biofuel cells (BFCs), allowing rapid glucose detection through dual signal transduction of current and electrochromic response. The anode (flexible Au/CNT-rGO paper films), functionalized with glucose oxidase/ferrocene (GOD/Fc), catalyzes glucose oxidation to gluconate and generates electrons and outputs the current signal, where Fc facilitates electron transport to mitigate oxygen dependency. The cathode (ITO/PET) provides a visual signal with chitosan@prussian (CS@PB) nano-composite integrated as an electrochromic region. The sensor exhibits a low detection limit of 0.018 mM and a linear detection range from 0.1 to 80 mM, with excellent selectivity and stability. In addition, clinical serum glucose and urine testing experiments validated the sensor's efficacy, demonstrating its potential for biomedical research and clinical applications. The sensor's self-power generation and dual-signal readout provide a promising platform for the development of point-of-care (POC) devices tailored for personalized metabolic health assessment.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"23 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144593870","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}
The demand for high-performance devices capable of both electromagnetic wave manipulation and biomedical detection has intensified with the growth of terahertz technologies. However, most existing devices are designed for a single function, either polarization control or biosensing which limits their utility in integrated systems. Addressing this challenge, we introduce a cost-effective terahertz device that seamlessly integrates broadband cross polarization transformation with precise refractive index sensing, all within a fabrication-friendly structure. Utilizing aluminum patterned resonators on a Rogers RT5870 dielectric layer, the device delivers a polarization conversion ratio above 94% across a wide spectral window of 3.492 THz with peak conversion efficiencies exceeding 99.9% at multiple resonances and retains stable performance at oblique incident angles up to 40°. Simultaneously, it functions as a refractive index sensor reaching a peak sensitivity of 1.35 THz/RIU and effectively distinguishes between healthy and diseased biological samples including blood, cervical and skin tissues. Compared to state-of-the-art designs which typically specialize in either polarization modulation or biosensing alone, the proposed device stands out for its dual functionality, high sensitivity and ultra-broadband performance. This work fills a rarely addressed research gap by delivering a single device capable of both advanced polarization control and reliable biomedical diagnostics, paving the way for multifunctional terahertz systems.
{"title":"Multifunctional Terahertz Device with Angular Resilience for Biomedical Sensing and Polarization Conversion","authors":"Lei Gao, Taha Sheheryar, Bo Lv","doi":"10.1039/d5an00646e","DOIUrl":"https://doi.org/10.1039/d5an00646e","url":null,"abstract":"The demand for high-performance devices capable of both electromagnetic wave manipulation and biomedical detection has intensified with the growth of terahertz technologies. However, most existing devices are designed for a single function, either polarization control or biosensing which limits their utility in integrated systems. Addressing this challenge, we introduce a cost-effective terahertz device that seamlessly integrates broadband cross polarization transformation with precise refractive index sensing, all within a fabrication-friendly structure. Utilizing aluminum patterned resonators on a Rogers RT5870 dielectric layer, the device delivers a polarization conversion ratio above 94% across a wide spectral window of 3.492 THz with peak conversion efficiencies exceeding 99.9% at multiple resonances and retains stable performance at oblique incident angles up to 40°. Simultaneously, it functions as a refractive index sensor reaching a peak sensitivity of 1.35 THz/RIU and effectively distinguishes between healthy and diseased biological samples including blood, cervical and skin tissues. Compared to state-of-the-art designs which typically specialize in either polarization modulation or biosensing alone, the proposed device stands out for its dual functionality, high sensitivity and ultra-broadband performance. This work fills a rarely addressed research gap by delivering a single device capable of both advanced polarization control and reliable biomedical diagnostics, paving the way for multifunctional terahertz systems.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"92 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144594833","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}
Retention time (RT) is essential in evaluating the likelihood of candidate structures in nontarget screening (NTS) with liquid chromatography high resolution mass spectrometry (LC/HRMS). Approaches for estimating RT of candidate structure can broadly be divided into projection and prediction methods. The first approach takes advantage of public databases of RTs measured on similar chromatographic systems (CSsource) and projects these to the chromatographic system applied in the NTS (CSNTS) based on a small set of commonly analyzed chemicals. The second approach leverages machine learning (ML) model(s) trained on publicly available retention time data measured on one or more chromatographic systems (CStraining). Nevertheless, the CSsource and CStraining might differ substantially from CSNTS. Therefore, it is of interest to evaluate the generalizability of projection models and prediction models in CSs routinely applied in NTS. Here we take advantage of the recent NORMAN interlaboratory comparison where 41 known calibration chemicals and 45 suspects were analyzed to evaluate both the projection and prediction approach on 37 CSs. The accuracy of both approaches was directly linked to the similarity of the CS and the pH of the mobile phase as well as the column chemistry were found to be most impactful. Furthermore, for cases where CSsource and CSNTS differ substantially but CStraining and CSNTS were similar, prediction models often performed on bar with the projection models. These findings highlight the need in accounting for the mobile phase and column chemistry in ML model training and selecting the prediction model for RT.
{"title":"Do Experimental Projection Methods Outcompete Retention Time Prediction Models in Non-target Screening? A Case Study on LC/HRMS Interlaboratory Comparison Data","authors":"Louise Malm, Anneli Kruve","doi":"10.1039/d5an00323g","DOIUrl":"https://doi.org/10.1039/d5an00323g","url":null,"abstract":"Retention time (RT) is essential in evaluating the likelihood of candidate structures in nontarget screening (NTS) with liquid chromatography high resolution mass spectrometry (LC/HRMS). Approaches for estimating RT of candidate structure can broadly be divided into projection and prediction methods. The first approach takes advantage of public databases of RTs measured on similar chromatographic systems (CS<small><sub>source</sub></small>) and projects these to the chromatographic system applied in the NTS (CS<small><sub>NTS</sub></small>) based on a small set of commonly analyzed chemicals. The second approach leverages machine learning (ML) model(s) trained on publicly available retention time data measured on one or more chromatographic systems (CS<small><sub>training</sub></small>). Nevertheless, the CS<small><sub>source</sub></small> and CS<small><sub>training</sub></small> might differ substantially from CSNTS. Therefore, it is of interest to evaluate the generalizability of projection models and prediction models in CSs routinely applied in NTS. Here we take advantage of the recent NORMAN interlaboratory comparison where 41 known calibration chemicals and 45 suspects were analyzed to evaluate both the projection and prediction approach on 37 CSs. The accuracy of both approaches was directly linked to the similarity of the CS and the pH of the mobile phase as well as the column chemistry were found to be most impactful. Furthermore, for cases where CS<small><sub>source</sub></small> and CS<small><sub>NTS</sub></small> differ substantially but CS<small><sub>training</sub></small> and CS<small><sub>NTS</sub></small> were similar, prediction models often performed on bar with the projection models. These findings highlight the need in accounting for the mobile phase and column chemistry in ML model training and selecting the prediction model for RT.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"105 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144577651","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}
Accompanied by the widespread use of antibiotics, antibiotic resistance genes (ARGs) have rapidly proliferated in the environment, necessitating the urgent development of sensitive detection methods. However, most reported methods focus on the determination of only one subtype of ARGs, which leads to an inadequate evaluation of ARG pollution levels. Surface-enhanced Raman scattering (SERS) assays are highly feasible and can serve as universal detection methods through Raman molecules. Conventional SERS assays may face background interference from complex matrices and demonstrate unsatisfactory sensitivity for trace analysis. In this study, a multifunctional magnetic recognition probe was designed to effectively separate and enrich two subtypes of penicillin ARGs. Subsequently, a cascade nicking-polymerization signal amplification process was successfully conducted for the captured ARGs, resulting in a SERS detection probe featuring two distinct Raman molecules capable of sensitive detection for bla-TEM and bla-CTX-M-1. This achievement can be likened to “hitting two birds with one stone”. Due to the signal amplification strategy, the developed sensor obtained good sensitivities with limits of detection (LOD) of 0.33 zM for bla-TEM and 0.29 zM for bla-CTX-M-1. Furthermore, the developed detection method enabled accurate quantitative analysis when compared with conventional polymerase chain reaction (PCR) assays.
{"title":"Raman Sensor Assisted by Cascade Signal Amplification for Simultaneous Detection of Two Extracellular Antibiotic Resistance Genes","authors":"Xinli Shi, Qi Wang, Yan Zhang, Chunping Ge, Yujun Sun, Yuqi Zhang, Mengmeng Li, Xiao Gong, Xin-Yue Song, Shusheng Zhang","doi":"10.1039/d5an00536a","DOIUrl":"https://doi.org/10.1039/d5an00536a","url":null,"abstract":"Accompanied by the widespread use of antibiotics, antibiotic resistance genes (ARGs) have rapidly proliferated in the environment, necessitating the urgent development of sensitive detection methods. However, most reported methods focus on the determination of only one subtype of ARGs, which leads to an inadequate evaluation of ARG pollution levels. Surface-enhanced Raman scattering (SERS) assays are highly feasible and can serve as universal detection methods through Raman molecules. Conventional SERS assays may face background interference from complex matrices and demonstrate unsatisfactory sensitivity for trace analysis. In this study, a multifunctional magnetic recognition probe was designed to effectively separate and enrich two subtypes of penicillin ARGs. Subsequently, a cascade nicking-polymerization signal amplification process was successfully conducted for the captured ARGs, resulting in a SERS detection probe featuring two distinct Raman molecules capable of sensitive detection for bla-TEM and bla-CTX-M-1. This achievement can be likened to “hitting two birds with one stone”. Due to the signal amplification strategy, the developed sensor obtained good sensitivities with limits of detection (LOD) of 0.33 zM for bla-TEM and 0.29 zM for bla-CTX-M-1. Furthermore, the developed detection method enabled accurate quantitative analysis when compared with conventional polymerase chain reaction (PCR) assays.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"107 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144577652","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}
In this study, we used three different potassium ionophores developed for ion-selective electrodes to fabricate nanoemulsion-type ion-selective optodes (NE-ISOs) with a pH-independent response mechanism based on cation exchange between a polarity-responsive cationic dye in organic phase and potassium ion in aqueous phase, and compared their responses. As a result, the NE-ISO prepared with valinomycin as the typical ionophore showed an ideal response in which the initial fluorescence intensity decreased with increasing ion concentration, as the cationic dye was extruded out into the aqueous phase following extraction of potassium ions by valinomycin. On the other hand, when the same experiment was performed using a bis-crown-type ionophore with a long alkyl chain, the initial fluorescence intensity became extremely small and increased with increasing potassium ion concentration, indicating a completely opposite response behavior that cannot be explained by the conventional ion exchange model. As a result of extraction experiments of hydrophobic organic cations and investigation of ionophore concentrations in nano droplet, we obtained some results that suggests that the interaction between the ionophore molecule and the dye molecule at the organic-aqueous interface and the accumulation behavior of the ion-ionophore complex at the organic-aqueous interface greatly affect the response behavior to ions, depending on the molecular structure of the ionophore. These results contradict the conventional perception in the development of ion-selective optodes that the same principle can be applied to measure different ions by changing the ionophore. It clearly demonstrates the importance of considering the chemical structures of the ionophore and dye molecules and their interaction, in addition to the response mechanism when designing NE-ISOs.
{"title":"Unusual response behavior of ion exchange-based potassium ion-selective nano-optodes based on bis(crown ether) neutral ionophores and cationic solvatochromic dye","authors":"Naoya Matsumoto, Kenji Sueyoshi, Tatsuro Endo, Hideaki Hisamoto","doi":"10.1039/d5an00516g","DOIUrl":"https://doi.org/10.1039/d5an00516g","url":null,"abstract":"In this study, we used three different potassium ionophores developed for ion-selective electrodes to fabricate nanoemulsion-type ion-selective optodes (NE-ISOs) with a pH-independent response mechanism based on cation exchange between a polarity-responsive cationic dye in organic phase and potassium ion in aqueous phase, and compared their responses. As a result, the NE-ISO prepared with valinomycin as the typical ionophore showed an ideal response in which the initial fluorescence intensity decreased with increasing ion concentration, as the cationic dye was extruded out into the aqueous phase following extraction of potassium ions by valinomycin. On the other hand, when the same experiment was performed using a bis-crown-type ionophore with a long alkyl chain, the initial fluorescence intensity became extremely small and increased with increasing potassium ion concentration, indicating a completely opposite response behavior that cannot be explained by the conventional ion exchange model. As a result of extraction experiments of hydrophobic organic cations and investigation of ionophore concentrations in nano droplet, we obtained some results that suggests that the interaction between the ionophore molecule and the dye molecule at the organic-aqueous interface and the accumulation behavior of the ion-ionophore complex at the organic-aqueous interface greatly affect the response behavior to ions, depending on the molecular structure of the ionophore. These results contradict the conventional perception in the development of ion-selective optodes that the same principle can be applied to measure different ions by changing the ionophore. It clearly demonstrates the importance of considering the chemical structures of the ionophore and dye molecules and their interaction, in addition to the response mechanism when designing NE-ISOs.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"31 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144569013","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}
Inertial microfluidics has demonstrated tremendous potential to impact biological - and notably medical - fields, by offering a highly versatile, portable and cost-effective approach to cell focusing and sorting. While the range of applications of inertial devices spans medical diagnostics, bioprocessing or water engineering to mention a few, translation is still impeded by the lack of clear understanding of cell interactions in such devices. This often leads to bespoke designs that take years of development and characterisation for one targeted application, and limited tools for informed optimisation. A more fundamental knowledge of inertial behaviours is key to future translational works and impact, by enabling a deeper understanding of inertial forces in biological systems. Towards this goal, this paper focuses on high-throughput morphological phenotyping of the single-celled, flagellated parasite Leishmania mexicana to better understand how variations in cell body length, width and flagellated status impact the focusing patterns of highly non-spherical cells in curved inertial devices. Some of the key findings in this study include i) flagella do not always alter focusing if body shape is conserved, ii) the impact of cell shape is specific to a channel design and slight changes in e.g., cell confinement can drastically change focusing patterns, iii) elongated prolate-like cells align differently depending on their lateral position within a curved channel, and iv) despite variabilities observed in focusing patterns for elongated versus rounder cell phenotypes, large morphological variations can be completely overcome at high Reynolds numbers so that all phenotypes tightly focus at a single and stable position (here, towards the channel outer wall). This last finding, in particular, may open new avenues for highly efficient cell enrichment processes.
{"title":"An experimental investigation into the focusing behaviours of flagellated and elongated cells in inertial microfluidic devices","authors":"Jessie Howell, Nicole Hall, Sulochana Omwenga, Tansy C. Hammarton, Melanie Jimenez","doi":"10.1039/d4an01288g","DOIUrl":"https://doi.org/10.1039/d4an01288g","url":null,"abstract":"Inertial microfluidics has demonstrated tremendous potential to impact biological - and notably medical - fields, by offering a highly versatile, portable and cost-effective approach to cell focusing and sorting. While the range of applications of inertial devices spans medical diagnostics, bioprocessing or water engineering to mention a few, translation is still impeded by the lack of clear understanding of cell interactions in such devices. This often leads to bespoke designs that take years of development and characterisation for one targeted application, and limited tools for informed optimisation. A more fundamental knowledge of inertial behaviours is key to future translational works and impact, by enabling a deeper understanding of inertial forces in biological systems. Towards this goal, this paper focuses on high-throughput morphological phenotyping of the single-celled, flagellated parasite Leishmania mexicana to better understand how variations in cell body length, width and flagellated status impact the focusing patterns of highly non-spherical cells in curved inertial devices. Some of the key findings in this study include i) flagella do not always alter focusing if body shape is conserved, ii) the impact of cell shape is specific to a channel design and slight changes in e.g., cell confinement can drastically change focusing patterns, iii) elongated prolate-like cells align differently depending on their lateral position within a curved channel, and iv) despite variabilities observed in focusing patterns for elongated versus rounder cell phenotypes, large morphological variations can be completely overcome at high Reynolds numbers so that all phenotypes tightly focus at a single and stable position (here, towards the channel outer wall). This last finding, in particular, may open new avenues for highly efficient cell enrichment processes.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"12 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144569016","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}
A contact co-culture model of bacteria and cells on the microfluidic chip with arrayed circular chambers was established to simulate the process of Salmonella typhimurium (S. typhimurium) invading HT-29 intestinal epithelial cells in this paper. The interaction process could be observed and detected in situ on the designed microchip. By optimizing the structure and size of chambers and microchannels, the microchip was more suitable for microscopic in-situ observation. The cell viability, morphology, cytoskeleton and secretions were detected during the interaction process of S. typhimurium invading HT-29 intestinal epithelial. In the experiments, cells formed multicellular aggregates, and cell cytoskeleton rearranged to form giant pinocytosis after exposure to S. typhimurium. It was shown that cell viability was continuously decreased during the process. Furthermore, it was also illustrated that cell viability was positively related to cytoskeleton damage and cell cytoskeleton regulated cell morphology. The cell secretions IL-8 in the co-culture supernatants were measured by enzyme-linked immunosorbent assay (ELISA). It was shown that the level of IL-8 increased rapidly when S. typhimurium invaded HT-29 intestinal epithelial cells, reached a peak of 4.45 pg/mL at 3 h, and then continuously came down until recovered to the baseline level of 1.04 pg/mL at 12 h. The designed microfluidic chip could provide a unique way to study the interaction of pathogen invaded cells and important information for the analysis of bacterial infection and other related research.
{"title":"Secretion analysis of invading process between Salmonella typhimurium and HT-29 intestinal epithelial cells on the microfluidic chip","authors":"Hong He, Haolan Tan, Chuang Ge, Yi Xu","doi":"10.1039/d5an00512d","DOIUrl":"https://doi.org/10.1039/d5an00512d","url":null,"abstract":"A contact co-culture model of bacteria and cells on the microfluidic chip with arrayed circular chambers was established to simulate the process of Salmonella typhimurium (S. typhimurium) invading HT-29 intestinal epithelial cells in this paper. The interaction process could be observed and detected in situ on the designed microchip. By optimizing the structure and size of chambers and microchannels, the microchip was more suitable for microscopic in-situ observation. The cell viability, morphology, cytoskeleton and secretions were detected during the interaction process of S. typhimurium invading HT-29 intestinal epithelial. In the experiments, cells formed multicellular aggregates, and cell cytoskeleton rearranged to form giant pinocytosis after exposure to S. typhimurium. It was shown that cell viability was continuously decreased during the process. Furthermore, it was also illustrated that cell viability was positively related to cytoskeleton damage and cell cytoskeleton regulated cell morphology. The cell secretions IL-8 in the co-culture supernatants were measured by enzyme-linked immunosorbent assay (ELISA). It was shown that the level of IL-8 increased rapidly when S. typhimurium invaded HT-29 intestinal epithelial cells, reached a peak of 4.45 pg/mL at 3 h, and then continuously came down until recovered to the baseline level of 1.04 pg/mL at 12 h. The designed microfluidic chip could provide a unique way to study the interaction of pathogen invaded cells and important information for the analysis of bacterial infection and other related research.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"109 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144569012","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}
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