Pub Date : 2024-11-01DOI: 10.1016/j.biosx.2024.100557
Dieter Spiehl , Gerhard Schwall , Fabian Post , Carina Weber , Edgar Dörsam , Andreas Blaeser , Volkhard A.J. Kempf , Michael Hogardt
This work presents a ready-to use dry culture device platform for differentiation of bacterial species and testing antibiotic susceptibility conveniently. The study demonstrates that high-volume and scalable production methods such as can reliably manufacture such devices for reproducible in vitro diagnostic testing. This marks a significant advancement compared to similar handmade devices in the literature.
Two types of ready-to-use (RTU) devices were produced through printing. One device differentiates bacterial species from urine samples for rapid diagnosis of urinary tract infections (UTI), while the other determines the minimum inhibitory concentration (MIC) of antibiotics against specific bacterial species. The results show performance comparable to standard tests in clinical microbiological laboratories. The differentiation of bacterial species from four UTI samples matched the efficacy of commercial UTI agar plates utilizing chromogenic substrates. The MIC determination for seven bacterial species in three independent tests yielded categorical results consistent with MIC test strips on agar plates as reference standard. Minor errors occurred in 14 % and major errors in 5 % of the tests. No major errors occurred and in 81 % of all tests results were correct and consistent with the reference method.
这项研究提出了一种可随时使用的干培养装置平台，用于区分细菌种类和方便地测试抗生素敏感性。该研究表明，大批量、可扩展的生产方法，如......，可以可靠地制造出这种装置，用于可重复的体外诊断检测。与文献中的类似手工设备相比，这标志着一项重大进步。通过印刷生产出了两种即用型设备（RTU）。其中一种装置可从尿液样本中区分细菌种类，用于快速诊断尿路感染（UTI）；另一种装置可确定抗生素对特定细菌种类的最小抑菌浓度（MIC）。结果显示，其性能可与临床微生物实验室的标准测试相媲美。从四份 UTI 样品中区分细菌种类的结果与使用显色底物的商用 UTI 琼脂平板的效果相当。在三次独立测试中对 7 种细菌进行了 MIC 测定，得出的分类结果与作为参考标准的琼脂平板上的 MIC 测试条一致。有 14% 的测试出现了轻微误差，5% 的测试出现了重大误差。所有测试中没有出现重大错误，81%的测试结果正确且与参考方法一致。
{"title":"Printed dry and ready-to-use in vitro diagnostic culture media devices for differentiation and antimicrobial susceptibility testing of bacteria","authors":"Dieter Spiehl , Gerhard Schwall , Fabian Post , Carina Weber , Edgar Dörsam , Andreas Blaeser , Volkhard A.J. Kempf , Michael Hogardt","doi":"10.1016/j.biosx.2024.100557","DOIUrl":"10.1016/j.biosx.2024.100557","url":null,"abstract":"<div><div>This work presents a ready-to use dry culture device platform for differentiation of bacterial species and testing antibiotic susceptibility conveniently. The study demonstrates that high-volume and scalable production methods such as can reliably manufacture such devices for reproducible in vitro diagnostic testing. This marks a significant advancement compared to similar handmade devices in the literature.</div><div>Two types of ready-to-use (RTU) devices were produced through printing. One device differentiates bacterial species from urine samples for rapid diagnosis of urinary tract infections (UTI), while the other determines the minimum inhibitory concentration (MIC) of antibiotics against specific bacterial species. The results show performance comparable to standard tests in clinical microbiological laboratories. The differentiation of bacterial species from four UTI samples matched the efficacy of commercial UTI agar plates utilizing chromogenic substrates. The MIC determination for seven bacterial species in three independent tests yielded categorical results consistent with MIC test strips on agar plates as reference standard. Minor errors occurred in 14 % and major errors in 5 % of the tests. No major errors occurred and in 81 % of all tests results were correct and consistent with the reference method.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100557"},"PeriodicalIF":10.61,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142587183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Invasive, biofilm-forming Non-typhoidal Salmonella (iNTS), propagating through the global food and water supply chain, presents a significant risk to food safety and public health. Developing a robust detection system is crucial for enabling point-of-care, affordable, and equipment-free identification of this pathogen throughout the supply chain. In this study, we screened a novel pool of ssDNA aptamers specific to a multidrug resistant iNTS strain SMC25, previously isolated from Indian poultry products in our earlier research. Through 13 rounds of whole-cell SELEX, we identified, characterized, and selected seven full-length aptamers (ST18, ST19, ST25, ST28, ST29, ST31, and ST32). Flow cytometric analysis reveals superior binding of ST25, ST28, ST29, and ST31. These aptamers were translated onto Nanozyme-based aptasensing system for efficient, cost-effective detection of SMC25. This system harnesses the aptamer-mediated, reversible peroxidase-like activity of gold nanoparticles (GNPs) to oxidize the TMB substrate into a one-electron oxidation state, resulting in a blue-colored Diamine charge transfer complex (DCTC). The catalytic process, coupled with GNP aggregation, induces a visible color change in the test mixture from ruby-red to blue. Post-SELEX truncations identified the optimal aptamer sequence (T_ST31), which selectively detected SMC25 in water with a limit of detection (LOD) of ∼10⁴ CFU/mL. Lower concentrations (10 CFU/mL) of SMC25 could be detected after non-selective enrichment within 120 min. This research introduces a novel pool of iNTS-specific aptamers along with a cost-effective (0.25 USD per sample) solution for colorimetric detection by the naked eye.
{"title":"Development of novel DNA aptamers and colorimetric nanozyme aptasensor for targeting multi-drug-resistant, invasive Salmonella typhimurium strain SMC25","authors":"Akanksha Joshi , Abhishek Kaushik , Neetu Kumra Taneja , Komal Chauhan , Amit Kumar , Tarun Kumar Sharma","doi":"10.1016/j.biosx.2024.100555","DOIUrl":"10.1016/j.biosx.2024.100555","url":null,"abstract":"<div><div>Invasive, biofilm-forming Non-typhoidal <em>Salmonella</em> (iNTS), propagating through the global food and water supply chain, presents a significant risk to food safety and public health. Developing a robust detection system is crucial for enabling point-of-care, affordable, and equipment-free identification of this pathogen throughout the supply chain. In this study, we screened a novel pool of ssDNA aptamers specific to a multidrug resistant iNTS strain SMC25, previously isolated from Indian poultry products in our earlier research. Through 13 rounds of whole-cell SELEX, we identified, characterized, and selected seven full-length aptamers (ST18, ST19, ST25, ST28, ST29, ST31, and ST32). Flow cytometric analysis reveals superior binding of ST25, ST28, ST29, and ST31. These aptamers were translated onto Nanozyme-based aptasensing system for efficient, cost-effective detection of SMC25. This system harnesses the aptamer-mediated, reversible peroxidase-like activity of gold nanoparticles (GNPs) to oxidize the TMB substrate into a one-electron oxidation state, resulting in a blue-colored Diamine charge transfer complex (DCTC). The catalytic process, coupled with GNP aggregation, induces a visible color change in the test mixture from ruby-red to blue. Post-SELEX truncations identified the optimal aptamer sequence (T_ST31), which selectively detected SMC25 in water with a limit of detection (LOD) of ∼10⁴ CFU/mL. Lower concentrations (10 CFU/mL) of SMC25 could be detected after non-selective enrichment within 120 min. This research introduces a novel pool of iNTS-specific aptamers along with a cost-effective (0.25 USD per sample) solution for colorimetric detection by the naked eye.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100555"},"PeriodicalIF":10.61,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142533623","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-19DOI: 10.1016/j.biosx.2024.100556
Tommi Åman , Sanna Auer , Vesa P. Hytönen , Juha A. Määttä
Biosensors are used in various applications in the field of medicine, environmental monitoring, and chemical processing to study the concentration and quality of target molecules. Biosensors convert the interaction between a specific target molecule and a recognition element into a measurable signal. We compared the performance of four different label-free biosensor techniques (multi-parametric surface plasmon resonance (MP-SPR), quartz crystal microbalance (QCM), mass-sensitive micro array (MSMA, or also known as film bulk acoustic resonators (FBARs)) and biolayer interferometry (BLI)) to evaluate how well they can quantify thick protein layers. We utilized the avidin-biotin system, which enables tight and specific binding and allowed us to assemble layers of proteins on the biosensor surface in a well-defined and reproducible fashion. Our results show that MP-SPR outperforms the other label-free biosensors in analyzing thick samples, showing a predictable and sensitive binding signal for over 50 albumin – avidin layers, which is estimated to correspond to a 300–400 nm thick protein layer. The linear measurement range of BLI was 38 layers corresponding to a 228–304 nm thick surface on the biosensor while QCM and MSMA were able to measure 108–144 nm and 72–96 nm thick protein layers with a fairly linear response, respectively.
{"title":"Performance of label-free biosensors as a function of layer thickness","authors":"Tommi Åman , Sanna Auer , Vesa P. Hytönen , Juha A. Määttä","doi":"10.1016/j.biosx.2024.100556","DOIUrl":"10.1016/j.biosx.2024.100556","url":null,"abstract":"<div><div>Biosensors are used in various applications in the field of medicine, environmental monitoring, and chemical processing to study the concentration and quality of target molecules. Biosensors convert the interaction between a specific target molecule and a recognition element into a measurable signal. We compared the performance of four different label-free biosensor techniques (multi-parametric surface plasmon resonance (MP-SPR), quartz crystal microbalance (QCM), mass-sensitive micro array (MSMA, or also known as film bulk acoustic resonators (FBARs)) and biolayer interferometry (BLI)) to evaluate how well they can quantify thick protein layers. We utilized the avidin-biotin system, which enables tight and specific binding and allowed us to assemble layers of proteins on the biosensor surface in a well-defined and reproducible fashion. Our results show that MP-SPR outperforms the other label-free biosensors in analyzing thick samples, showing a predictable and sensitive binding signal for over 50 albumin – avidin layers, which is estimated to correspond to a 300–400 nm thick protein layer. The linear measurement range of BLI was 38 layers corresponding to a 228–304 nm thick surface on the biosensor while QCM and MSMA were able to measure 108–144 nm and 72–96 nm thick protein layers with a fairly linear response, respectively.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100556"},"PeriodicalIF":10.61,"publicationDate":"2024-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142533622","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-17DOI: 10.1016/j.biosx.2024.100554
Simonetta Palleschi , Leopoldo Silvestroni , Barbara Rossi , Simone Dinarelli , Marco Magi , Lorenzo Giacomelli , Andrea Bettucci
Viscoelasticity (VE) is the intrinsic mechano-dynamic property enabling red blood cells (RBCs) to undergo prompt and repeated deformations while maintaining structural integrity. Assessing RBC VE and how different stressors can affect it is of great interest. Quartz Crystal Microbalance with Dissipation monitoring (QCM-D) is a technology exploiting high-frequency acoustic waves to probe soft matter rheological properties. In the present study, a QCM-D method is reported for in vitro monitoring of cell VE in viable RBCs. The method is based on casting a sensor-adherent cell monolayer and modeling it as an effective viscoelastic medium, and allows to extrapolate proxy values of both the elastic and the viscous cell shear moduli. Real-time VE changes induced by the known cell VE stressors temperature, medium tonicity, glutaraldehyde, methyl-β-cyclodextrin and cytochalasin D have been reliably identified. The method is relatively simple and inexpensive, non-invasive, and able to seize subtle changes of cell biomechanics. Hence, it could be usefully exploited for in vitro assessment of RBC rheological properties and their alterations induced by external chemico-physical stimuli.
粘弹性（VE）是红细胞（RBC）在保持结构完整性的同时能够迅速、反复变形的内在机械动力特性。评估 RBC 的 VE 以及不同的应力如何影响它是非常有意义的。具有耗散监测功能的石英晶体微天平（QCM-D）是一种利用高频声波探测软物质流变特性的技术。本研究报告了一种用于体外监测存活 RBC 中细胞 VE 的 QCM-D 方法。该方法基于铸造一个传感器附着细胞单层，并将其建模为有效粘弹性介质，从而推断出细胞弹性和粘性剪切模量的代理值。已知的细胞粘弹性应激源温度、培养基补液度、戊二醛、甲基-β-环糊精和细胞松弛素 D 引起的实时粘弹性变化已被可靠地识别出来。该方法相对简单、廉价、无创，并能捕捉到细胞生物力学的微妙变化。因此，该方法可用于体外评估 RBC 流变特性及其在外部化学物理刺激下的变化。
{"title":"Simple and sensitive method for in vitro monitoring of red blood cell viscoelasticity by Quartz Crystal Microbalance with dissipation monitoring (QCM-D)","authors":"Simonetta Palleschi , Leopoldo Silvestroni , Barbara Rossi , Simone Dinarelli , Marco Magi , Lorenzo Giacomelli , Andrea Bettucci","doi":"10.1016/j.biosx.2024.100554","DOIUrl":"10.1016/j.biosx.2024.100554","url":null,"abstract":"<div><div>Viscoelasticity (VE) is the intrinsic mechano-dynamic property enabling red blood cells (RBCs) to undergo prompt and repeated deformations while maintaining structural integrity. Assessing RBC VE and how different stressors can affect it is of great interest. Quartz Crystal Microbalance with Dissipation monitoring (QCM-D) is a technology exploiting high-frequency acoustic waves to probe soft matter rheological properties. In the present study, a QCM-D method is reported for <em>in vitro</em> monitoring of cell VE in viable RBCs. The method is based on casting a sensor-adherent cell monolayer and modeling it as an effective viscoelastic medium, and allows to extrapolate proxy values of both the elastic and the viscous cell shear moduli. Real-time VE changes induced by the known cell VE stressors temperature, medium tonicity, glutaraldehyde, methyl-β-cyclodextrin and cytochalasin D have been reliably identified. The method is relatively simple and inexpensive, non-invasive, and able to seize subtle changes of cell biomechanics. Hence, it could be usefully exploited for <em>in vitro</em> assessment of RBC rheological properties and their alterations induced by external chemico-physical stimuli.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100554"},"PeriodicalIF":10.61,"publicationDate":"2024-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142533621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-10DOI: 10.1016/j.biosx.2024.100551
Shengdong Li
Lipid droplets participate in the metabolic process of many living cells, and their abnormalities are highly correlated with many diseases such as fatty liver, diabetes, and cancer. Nevertheless, the current methods are inadequate for elucidating the relationship between diverse diseases and lipid droplets. In particular, the visualization of tumors with abnormal distribution of lipid droplets remains a significant challenge. Therefore, a water-soluble biosensor with lipid droplet specificity is synthesized by copolymerization of coumarin derivatives with low toxicity and high hydrophilic hydroxyl terminal polyethylene glycol. The survival rate of HeLa cells cultured with lipid drop biosensor for 24 h was more than 90% on average, and the toxicity was less. The co-localization imaging experiments with lipid droplets showed that the biosensor had a good ability to target lipid droplets in HeLa cells. The experimental results of the response of different cell lines to the biosensor showed that the tumor cells had a good response and uptake ability. These results indicate that the prepared biosensor has a good distinguishing function between normal cells and tumor cells, and provides an experimental basis for the early diagnosis of clinical tumors.
脂滴参与许多活细胞的新陈代谢过程，其异常与脂肪肝、糖尿病和癌症等多种疾病高度相关。然而，目前的方法还不足以阐明各种疾病与脂滴之间的关系。特别是，如何观察脂滴分布异常的肿瘤仍然是一个重大挑战。因此，通过低毒性香豆素衍生物与高亲水性羟基端聚乙二醇的共聚，合成了一种具有脂滴特异性的水溶性生物传感器。用脂滴生物传感器培养 HeLa 细胞 24 小时的存活率平均超过 90%，且毒性较低。脂滴共定位成像实验表明，该生物传感器能很好地靶向 HeLa 细胞中的脂滴。不同细胞系对生物传感器的反应实验结果表明，肿瘤细胞对生物传感器具有良好的反应和吸收能力。这些结果表明，制备的生物传感器具有良好的区分正常细胞和肿瘤细胞的功能，为临床肿瘤的早期诊断提供了实验依据。
{"title":"Targeted biosensors for intracellular lipid droplet content detection","authors":"Shengdong Li","doi":"10.1016/j.biosx.2024.100551","DOIUrl":"10.1016/j.biosx.2024.100551","url":null,"abstract":"<div><div>Lipid droplets participate in the metabolic process of many living cells, and their abnormalities are highly correlated with many diseases such as fatty liver, diabetes, and cancer. Nevertheless, the current methods are inadequate for elucidating the relationship between diverse diseases and lipid droplets. In particular, the visualization of tumors with abnormal distribution of lipid droplets remains a significant challenge. Therefore, a water-soluble biosensor with lipid droplet specificity is synthesized by copolymerization of coumarin derivatives with low toxicity and high hydrophilic hydroxyl terminal polyethylene glycol. The survival rate of HeLa cells cultured with lipid drop biosensor for 24 h was more than 90% on average, and the toxicity was less. The co-localization imaging experiments with lipid droplets showed that the biosensor had a good ability to target lipid droplets in HeLa cells. The experimental results of the response of different cell lines to the biosensor showed that the tumor cells had a good response and uptake ability. These results indicate that the prepared biosensor has a good distinguishing function between normal cells and tumor cells, and provides an experimental basis for the early diagnosis of clinical tumors.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100551"},"PeriodicalIF":10.61,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142433225","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-10DOI: 10.1016/j.biosx.2024.100552
Jefferson Honorio Franco , Paolo Stufano , Rossella Labarile , Dario Lacalamita , Pierluigi Lasala , Elisabetta Fanizza , Massimo Trotta , Gianluca Maria Farinola , Matteo Grattieri
The low-cost and early monitoring of metal ion contaminants is paramount to prevent widespread contamination of water environments. Self-powered microbial electrochemical sensors represent an interesting approach to achieving this goal. Purple non-sulfur bacteria have a versatile metabolism and a well-characterized photosynthetic system, making them an ideal candidate for developing biohybrid technologies. In this work, we report the use of these bacteria in biophotoelectrodes to develop self-powered monitoring systems for two common pollutants, NiCl2 and CuSO4. The microbial biophotoelectrode was obtained on a homemade poly-hydroxybutyrate-carbon nanofibers electrode modified with a redox-adhesive polydopamine matrix-based entrapping the purple bacterium Rhodobacter capsulatus. The presence of 500 μM NiCl2 resulted in a 60 % decrease in current density, while the simultaneous presence of 100 μM NiCl2 and 100 mM CuSO4 led to an 83 % current inhibition. Given the implementation of the biophotoelectrode in the field, the biohybrid system was tested in a complex matrix containing beer, demonstrating the promising ability of the photoelectrochemical system to act as an efficient biosensor in complex solutions. Finally, the biohybrid electrode was coupled to a cathode performing oxygen reduction, which allowed obtaining a self-powered monitoring system, paving the way for the future implementation of a low-cost monitoring system for widespread metal ions contaminant monitoring.
{"title":"Intact photosynthetic bacteria-based electrodes for self-powered metal ions monitoring","authors":"Jefferson Honorio Franco , Paolo Stufano , Rossella Labarile , Dario Lacalamita , Pierluigi Lasala , Elisabetta Fanizza , Massimo Trotta , Gianluca Maria Farinola , Matteo Grattieri","doi":"10.1016/j.biosx.2024.100552","DOIUrl":"10.1016/j.biosx.2024.100552","url":null,"abstract":"<div><div>The low-cost and early monitoring of metal ion contaminants is paramount to prevent widespread contamination of water environments. Self-powered microbial electrochemical sensors represent an interesting approach to achieving this goal. Purple non-sulfur bacteria have a versatile metabolism and a well-characterized photosynthetic system, making them an ideal candidate for developing biohybrid technologies. In this work, we report the use of these bacteria in biophotoelectrodes to develop self-powered monitoring systems for two common pollutants, NiCl<sub>2</sub> and CuSO<sub>4</sub>. The microbial biophotoelectrode was obtained on a homemade poly-hydroxybutyrate-carbon nanofibers electrode modified with a redox-adhesive polydopamine matrix-based entrapping the purple bacterium <em>Rhodobacter capsulatus</em>. The presence of 500 μM NiCl<sub>2</sub> resulted in a 60 % decrease in current density, while the simultaneous presence of 100 μM NiCl<sub>2</sub> and 100 mM CuSO<sub>4</sub> led to an 83 % current inhibition. Given the implementation of the biophotoelectrode in the field, the biohybrid system was tested in a complex matrix containing beer, demonstrating the promising ability of the photoelectrochemical system to act as an efficient biosensor in complex solutions. Finally, the biohybrid electrode was coupled to a cathode performing oxygen reduction, which allowed obtaining a self-powered monitoring system, paving the way for the future implementation of a low-cost monitoring system for widespread metal ions contaminant monitoring.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100552"},"PeriodicalIF":10.61,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142433226","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study introduces a novel tyrosinase-based biosensor designed to simultaneously detect tyrosinase, tyrosine, L-DOPA, and L-DOPA quinone. The biosensor, designated as GCE/GNP/Cys/Chit/Tyrase, was developed by immobilizing tyrosinase on a modified glassy carbon electrode (GCE) that incorporates electrodeposited gold nanoparticles, cysteine, and chitosan. The morphology of the biosensor was characterized using scanning electron microscopy (SEM). The electrochemical behaviors of the biosensor were explored in response to the target analytes. Key analytical characteristics were assessed, including linear range, sensitivity, selectivity, limits of detection and quantification, long-term stability, repeatability, reproducibility, electrochemically active surface area, and charge transfer behavior. The biosensor demonstrated a linear response range of 1–120 μM, with an impressive sensitivity of 200.4 mA.Lmol−1cm−2 and a detection limit of 27 μM. This study provides a comprehensive evaluation of the analytical features of tyrosinase-based biosensors. Additionally, the biosensor was applied to quantify dopamine in brain tissue, utilizing a calibration curve derived from the fabricated biosensor. The dopamine concentration measured in five Wistar rats was 35 ± 2.75 μM, reflecting the mean and standard deviation, respectively. These results confirm the biosensor's capability for accurately detecting this critical neurotransmitter in the brains of Wistar rats. This investigation underscores the potential of tyrosinase-based biosensors for diverse analytical applications in biological samples.
{"title":"A novel approach to tyrosinase-based biosensors: Electrode reactions and biological measurement","authors":"Masumeh Goodarzi , Hosna Tavakoli , Maryam Arab Khalaj , Hassan Tavakoli","doi":"10.1016/j.biosx.2024.100550","DOIUrl":"10.1016/j.biosx.2024.100550","url":null,"abstract":"<div><div>This study introduces a novel tyrosinase-based biosensor designed to simultaneously detect tyrosinase, tyrosine, L-DOPA, and L-DOPA quinone. The biosensor, designated as GCE/GNP/Cys/Chit/Tyrase, was developed by immobilizing tyrosinase on a modified glassy carbon electrode (GCE) that incorporates electrodeposited gold nanoparticles, cysteine, and chitosan. The morphology of the biosensor was characterized using scanning electron microscopy (SEM). The electrochemical behaviors of the biosensor were explored in response to the target analytes. Key analytical characteristics were assessed, including linear range, sensitivity, selectivity, limits of detection and quantification, long-term stability, repeatability, reproducibility, electrochemically active surface area, and charge transfer behavior. The biosensor demonstrated a linear response range of 1–120 μM, with an impressive sensitivity of 200.4 mA.Lmol<sup>−</sup><sup>1</sup>cm<sup>−</sup><sup>2</sup> and a detection limit of 27 μM. This study provides a comprehensive evaluation of the analytical features of tyrosinase-based biosensors. Additionally, the biosensor was applied to quantify dopamine in brain tissue, utilizing a calibration curve derived from the fabricated biosensor. The dopamine concentration measured in five Wistar rats was 35 ± 2.75 μM, reflecting the mean and standard deviation, respectively. These results confirm the biosensor's capability for accurately detecting this critical neurotransmitter in the brains of Wistar rats. This investigation underscores the potential of tyrosinase-based biosensors for diverse analytical applications in biological samples.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100550"},"PeriodicalIF":10.61,"publicationDate":"2024-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142417340","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-05DOI: 10.1016/j.biosx.2024.100549
Zuyuan Tian , Mohamed Shaheen , Tianxiang Jiang , Shaoxi Wang , Xihua Wang , Jie Chen
Rapid isolation and precise quantification of target cancer cells are crucial for precision medicine applications. Conventional fluorescence-based methods require bulky optical instrumentation and specialized expertise. Recent advances in label-free microfluidic techniques have attempted to overcome these limitations but often suffer from reduced reliability and throughput when integrating multiple functions. Here, we demonstrate an electroactive microfluidic platform that integrates functions of selective concentration, particle focusing, and single-cell level quantification without introducing additional complex physical microstructures. Our device employs the dielectrophoresis (DEP) effect combined with an interdigitated cell sorter to achieve selective cell concentration. For quantification and characterization, the device leverages the impedimetric Coulter principle to achieve precise particle counting and characterization at single-cell resolution. To address the inherent tradeoff between sensitivity and throughput in the traditional Coulter counter, we utilize the tilted interdigitated electrode configuration excited by an amplitude-modulated (AM) Positive DEP (pDEP) signal instead of the commonly employed narrow channel or sheath flow to realize the effective focusing and alignment of the cell stream. The focusing module is combined with our side-counter design to detect cells. The performance of our device and its embedded modules has been verified with mixed breast cancer and blood model cell lines, where high consistency between optical and electrical detection has been demonstrated. We expect the integration of the proposed AM-pDEP focusing approach and the corresponding microfluidic design for cell sorter and on-chip flow cytometry would offer an alternative engineering solution to the stream-based on-chip single-cell analysis where reliable cell focusing and single-cell sensing are required.
{"title":"An electroactive microfluidic platform integrated with AM-pDEP focusing and side-counter design for selective cell sorting and single-cell quantification","authors":"Zuyuan Tian , Mohamed Shaheen , Tianxiang Jiang , Shaoxi Wang , Xihua Wang , Jie Chen","doi":"10.1016/j.biosx.2024.100549","DOIUrl":"10.1016/j.biosx.2024.100549","url":null,"abstract":"<div><div>Rapid isolation and precise quantification of target cancer cells are crucial for precision medicine applications. Conventional fluorescence-based methods require bulky optical instrumentation and specialized expertise. Recent advances in label-free microfluidic techniques have attempted to overcome these limitations but often suffer from reduced reliability and throughput when integrating multiple functions. Here, we demonstrate an electroactive microfluidic platform that integrates functions of selective concentration, particle focusing, and single-cell level quantification without introducing additional complex physical microstructures. Our device employs the dielectrophoresis (DEP) effect combined with an interdigitated cell sorter to achieve selective cell concentration. For quantification and characterization, the device leverages the impedimetric Coulter principle to achieve precise particle counting and characterization at single-cell resolution. To address the inherent tradeoff between sensitivity and throughput in the traditional Coulter counter, we utilize the tilted interdigitated electrode configuration excited by an amplitude-modulated (AM) Positive DEP (pDEP) signal instead of the commonly employed narrow channel or sheath flow to realize the effective focusing and alignment of the cell stream. The focusing module is combined with our side-counter design to detect cells. The performance of our device and its embedded modules has been verified with mixed breast cancer and blood model cell lines, where high consistency between optical and electrical detection has been demonstrated. We expect the integration of the proposed AM-pDEP focusing approach and the corresponding microfluidic design for cell sorter and on-chip flow cytometry would offer an alternative engineering solution to the stream-based on-chip single-cell analysis where reliable cell focusing and single-cell sensing are required.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100549"},"PeriodicalIF":10.61,"publicationDate":"2024-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142417344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study used a polyurethane (PU)-based ion selective electrode (ISE) immobilized with di-(2-ethyl hexyl) phosphoric acid (D2EHPA) to detect and quantify Pb2+ ions at low concentrations (10−10 to 10−1 M) in aqueous solution. PU, synthesized from castor oil (Ricinus communis L.), was utilized as the ISE membrane matrix. The ISE demonstrated a sensitivity of 26.24 ± 0.12 mV/decade, a detection limit of 1.44 × 10−7 M, and a response time of 10 s. We maintained the measurement stability for up to six days of storage.
The results of Pb2+ quantification produced by ISE were compared with the results using atomic absorption spectroscopy (AAS), indicating similar Pb2+ concentrations in both artificial and real wastewater samples (calculated t-value < theoretical t-value). Therefore, this ISE is suitable for detecting trace levels of Pb2+ and quantifying them with high accuracy.
本研究使用固定了二-（2-乙基己基）磷酸（D2EHPA）的聚氨酯（PU）基离子选择电极（ISE）来检测和定量水溶液中低浓度（10-10 至 10-1 M）的 Pb2+ 离子。用蓖麻油（Ricinus communis L.）合成的聚氨酯作为 ISE 膜基质。ISE 的灵敏度为 26.24 ± 0.12 mV/decade，检测限为 1.44 × 10-7 M，响应时间为 10 s。我们将 ISE 的 Pb2+ 定量结果与原子吸收光谱法（AAS）的结果进行了比较，结果表明人工废水和实际废水样品中的 Pb2+ 浓度相似（计算 t 值 < 理论 t 值）。因此，这种 ISE 适用于检测痕量 Pb2+，并能准确定量。
{"title":"Ion-selective electrode based on polyurethane-immobilized di-(2-ethyl hexyl) phosphoric acid for low-concentration aqueous Pb2+ detection and quantification","authors":"Khairun Nisah , Eka Safitri , Rahmi Rahmi , Muliadi Ramli , Reni Silvia Nasution , Muhammad Iqhrammullah","doi":"10.1016/j.biosx.2024.100547","DOIUrl":"10.1016/j.biosx.2024.100547","url":null,"abstract":"<div><div>This study used a polyurethane (PU)-based ion selective electrode (ISE) immobilized with di-(2-ethyl hexyl) phosphoric acid (D2EHPA) to detect and quantify Pb<sup>2+</sup> ions at low concentrations (10<sup>−10</sup> to 10<sup>−1</sup> M) in aqueous solution. PU, synthesized from castor oil (<em>Ricinus communis</em> L.), was utilized as the ISE membrane matrix. The ISE demonstrated a sensitivity of 26.24 ± 0.12 mV/decade, a detection limit of 1.44 × 10<sup>−7</sup> M, and a response time of 10 s. We maintained the measurement stability for up to six days of storage.</div><div>The results of Pb<sup>2+</sup> quantification produced by ISE were compared with the results using atomic absorption spectroscopy (AAS), indicating similar Pb<sup>2+</sup> concentrations in both artificial and real wastewater samples (calculated t-value < theoretical t-value). Therefore, this ISE is suitable for detecting trace levels of Pb<sup>2+</sup> and quantifying them with high accuracy.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100547"},"PeriodicalIF":10.61,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142433242","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-24DOI: 10.1016/j.biosx.2024.100546
Hengye Huang , Karol Kexin Sun , Dan Wu , Chuncao Zhang , Yanyan Huo , Yuan Tian , Guangjun Yu
Global developmental delay (GDD) is a common neurodevelopmental disorder in children under 5 years of age. This study assessed the potential of functional Near-Infrared Spectroscopy (fNIRS) technology aiding diagnosis of GDD in premature infants and explored brain pathogenesis of GDD. This prospective cohort study was conducted between December 2020 and June 2023 among preterm infants in five hospitals in China. The primary outcome was the diagnosis of GDD. A 5-min fNIRS brain scan was performed in preterm infants at 4-, 8-, and 12-month corrected age in two hospitals. A diagnostic model based on fNIRS brain network and an integrated diagnostic model built on fNIRS data as well as birth head circumference was developed. Diagnostic models were assessed using the area under the curve (AUC) from receiver operating characteristic curves. Brain network analysis revealed significantly worse left-sided functional connectivity in GDD preterm infants. The fNIRS technique had satisfactory diagnostic value for GDD in all age groups. The diagnostic accuracy of the combined diagnostic model significantly improved. This study suggested that fNIRS could be considered for early screening and late auxiliary diagnosis of GDD in preterm infants. Furthermore, it can also be used to explore GDD brain pathogenesis.
{"title":"Resting-state brain network analysis and applied evaluation of global developmental delay in preterm infants: A functional near-infrared spectroscopic study","authors":"Hengye Huang , Karol Kexin Sun , Dan Wu , Chuncao Zhang , Yanyan Huo , Yuan Tian , Guangjun Yu","doi":"10.1016/j.biosx.2024.100546","DOIUrl":"10.1016/j.biosx.2024.100546","url":null,"abstract":"<div><div>Global developmental delay (GDD) is a common neurodevelopmental disorder in children under 5 years of age. This study assessed the potential of functional Near-Infrared Spectroscopy (fNIRS) technology aiding diagnosis of GDD in premature infants and explored brain pathogenesis of GDD. This prospective cohort study was conducted between December 2020 and June 2023 among preterm infants in five hospitals in China. The primary outcome was the diagnosis of GDD. A 5-min fNIRS brain scan was performed in preterm infants at 4-, 8-, and 12-month corrected age in two hospitals. A diagnostic model based on fNIRS brain network and an integrated diagnostic model built on fNIRS data as well as birth head circumference was developed. Diagnostic models were assessed using the area under the curve (AUC) from receiver operating characteristic curves. Brain network analysis revealed significantly worse left-sided functional connectivity in GDD preterm infants. The fNIRS technique had satisfactory diagnostic value for GDD in all age groups. The diagnostic accuracy of the combined diagnostic model significantly improved. This study suggested that fNIRS could be considered for early screening and late auxiliary diagnosis of GDD in preterm infants. Furthermore, it can also be used to explore GDD brain pathogenesis.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"21 ","pages":"Article 100546"},"PeriodicalIF":10.61,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142653619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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