Sven Meinen, Steffen Brinkmann, Kevin Viebrock, Bassant Elbardisy, Henning Menzel, Rainer Krull, Andreas Dietzel
Microbioreactors increase information output in biopharmaceutical screening applications because they can be operated in parallel without consuming large quantities of the pharmaceutical formulations being tested. A capillary wave microbioreactor (cwMBR) has recently been reported, allowing cost-efficient parallelization in an array that can be activated for mixing as a whole. Although impedance spectroscopy can directly distinguish between dead and viable cells, the monitoring of cells in suspension within bioreactors is challenging because the signal is influenced by the potentially varying properties of the culture medium. In order to address this challenge, an impedance sensor consisting of two sets of microelectrodes in a cwMBR is presented. Only one set of electrodes was covered by a two-photon cross-linked hydrogel to become insensitive to the influence of cells while remaining sensitive to the culture medium. With this impedance sensor, the biomass of Saccharomyces cerevisiae could be measured in a range from 1 to 20g/l. In addition, the sensor can compensate for a change in the conductivity of the suspension of 5 to 15mS/cm. Moreover, the two-photon cross-linking of hydroxyethyl starch methacrylate hydrogel, which has been studied in detail, recommends itself for even much broader sensing applications in miniaturized bioreactors and biosensors.
{"title":"2PP-Hydrogel Covered Electrodes to Compensate for Media Effects in the Determination of Biomass in a Capillary Wave Micro Bioreactor","authors":"Sven Meinen, Steffen Brinkmann, Kevin Viebrock, Bassant Elbardisy, Henning Menzel, Rainer Krull, Andreas Dietzel","doi":"10.3390/bios14090438","DOIUrl":"https://doi.org/10.3390/bios14090438","url":null,"abstract":"Microbioreactors increase information output in biopharmaceutical screening applications because they can be operated in parallel without consuming large quantities of the pharmaceutical formulations being tested. A capillary wave microbioreactor (cwMBR) has recently been reported, allowing cost-efficient parallelization in an array that can be activated for mixing as a whole. Although impedance spectroscopy can directly distinguish between dead and viable cells, the monitoring of cells in suspension within bioreactors is challenging because the signal is influenced by the potentially varying properties of the culture medium. In order to address this challenge, an impedance sensor consisting of two sets of microelectrodes in a cwMBR is presented. Only one set of electrodes was covered by a two-photon cross-linked hydrogel to become insensitive to the influence of cells while remaining sensitive to the culture medium. With this impedance sensor, the biomass of Saccharomyces cerevisiae could be measured in a range from 1 to 20g/l. In addition, the sensor can compensate for a change in the conductivity of the suspension of 5 to 15mS/cm. Moreover, the two-photon cross-linking of hydroxyethyl starch methacrylate hydrogel, which has been studied in detail, recommends itself for even much broader sensing applications in miniaturized bioreactors and biosensors.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"19 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Plasmonic intragap nanostructures (PINs) have garnered intensive attention in Raman-related analysis due to their exceptional ability to enhance light–matter interactions. Although diverse synthetic strategies have been employed to create these nanostructures, the emphasis has largely been on PINs with simple configurations, which often fall short in achieving effective near-field focusing. Three-dimensional (3D) complex PINs, distinguished by their intricate networks of internal gaps and voids, are emerging as superior structures for effective light trapping. These structures facilitate the generation of hot spots and hot zones that are essential for enhanced near-field focusing. Nevertheless, the synthesis techniques for these complex structures and their specific impacts on near-field focusing are not well-documented. This review discusses the recent advancements in the synthesis of 3D complex PINs and their applications in surface-enhanced Raman scattering (SERS). We begin by describing the foundational methods for fabricating simple PINs, followed by a discussion on the rational design strategies aimed at developing 3D complex PINs with superior near-field focusing capabilities. We also evaluate the SERS performance of various 3D complex PINs, emphasizing their advanced sensing capabilities. Lastly, we explore the future perspective of 3D complex PINs in SERS applications.
{"title":"Recent Progress in the Synthesis of 3D Complex Plasmonic Intragap Nanostructures and Their Applications in Surface-Enhanced Raman Scattering","authors":"Li Ma, Keyi Zhou, Xinyue Wang, Jiayue Wang, Ruyu Zhao, Yifei Zhang, Fang Cheng","doi":"10.3390/bios14090433","DOIUrl":"https://doi.org/10.3390/bios14090433","url":null,"abstract":"Plasmonic intragap nanostructures (PINs) have garnered intensive attention in Raman-related analysis due to their exceptional ability to enhance light–matter interactions. Although diverse synthetic strategies have been employed to create these nanostructures, the emphasis has largely been on PINs with simple configurations, which often fall short in achieving effective near-field focusing. Three-dimensional (3D) complex PINs, distinguished by their intricate networks of internal gaps and voids, are emerging as superior structures for effective light trapping. These structures facilitate the generation of hot spots and hot zones that are essential for enhanced near-field focusing. Nevertheless, the synthesis techniques for these complex structures and their specific impacts on near-field focusing are not well-documented. This review discusses the recent advancements in the synthesis of 3D complex PINs and their applications in surface-enhanced Raman scattering (SERS). We begin by describing the foundational methods for fabricating simple PINs, followed by a discussion on the rational design strategies aimed at developing 3D complex PINs with superior near-field focusing capabilities. We also evaluate the SERS performance of various 3D complex PINs, emphasizing their advanced sensing capabilities. Lastly, we explore the future perspective of 3D complex PINs in SERS applications.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"33 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186546","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jun Guo, Xuanqi Wang, Ruiyu Bai, Zimo Zhang, Huazhen Chen, Kai Xue, Chuang Ma, Dawei Zang, Erwei Yin, Kunpeng Gao, Bowen Ji
Compared with the traditional gel electrode, the dry electrode is being taken more seriously in bioelectrical recording because of its easy preparation, long-lasting ability, and reusability. However, the commonly used dry AgCl electrodes and silver cloth electrodes are generally hard to record through hair due to their flat contact surface. Claw electrodes can contact skin through hair on the head and body, but the internal claw structure is relatively hard and causes discomfort after being worn for a few hours. Here, we report a conductive Velcro electrode (CVE) with an elastic hook hair structure, which can collect biopotential through body hair. The elastic hooks greatly reduce discomfort after long-time wearing and can even be worn all day. The CVE electrode is fabricated by one-step immersion in conductive silver paste based on the cost-effective commercial Velcro, forming a uniform and durable conductive coating on a cluster of hook microstructures. The electrode shows excellent properties, including low impedance (15.88 kΩ @ 10 Hz), high signal-to-noise ratio (16.0 dB), strong water resistance, and mechanical resistance. After washing in laundry detergent, the impedance of CVE is still 16% lower than the commercial AgCl electrodes. To verify the mechanical strength and recovery capability, we conducted cyclic compression experiments. The results show that the displacement change of the electrode hook hair after 50 compression cycles was still less than 1%. This electrode provides a universal acquisition scheme, including effective acquisition of different parts of the body with or without hair. Finally, the gesture recognition from electromyography (EMG) by the CVE electrode was applied with accuracy above 90%. The CVE proposed in this study has great potential and promise in various human–machine interface (HMI) applications that employ surface biopotential signals on the body or head with hair.
{"title":"A Cost-Effective and Easy-to-Fabricate Conductive Velcro Dry Electrode for Durable and High-Performance Biopotential Acquisition","authors":"Jun Guo, Xuanqi Wang, Ruiyu Bai, Zimo Zhang, Huazhen Chen, Kai Xue, Chuang Ma, Dawei Zang, Erwei Yin, Kunpeng Gao, Bowen Ji","doi":"10.3390/bios14090432","DOIUrl":"https://doi.org/10.3390/bios14090432","url":null,"abstract":"Compared with the traditional gel electrode, the dry electrode is being taken more seriously in bioelectrical recording because of its easy preparation, long-lasting ability, and reusability. However, the commonly used dry AgCl electrodes and silver cloth electrodes are generally hard to record through hair due to their flat contact surface. Claw electrodes can contact skin through hair on the head and body, but the internal claw structure is relatively hard and causes discomfort after being worn for a few hours. Here, we report a conductive Velcro electrode (CVE) with an elastic hook hair structure, which can collect biopotential through body hair. The elastic hooks greatly reduce discomfort after long-time wearing and can even be worn all day. The CVE electrode is fabricated by one-step immersion in conductive silver paste based on the cost-effective commercial Velcro, forming a uniform and durable conductive coating on a cluster of hook microstructures. The electrode shows excellent properties, including low impedance (15.88 kΩ @ 10 Hz), high signal-to-noise ratio (16.0 dB), strong water resistance, and mechanical resistance. After washing in laundry detergent, the impedance of CVE is still 16% lower than the commercial AgCl electrodes. To verify the mechanical strength and recovery capability, we conducted cyclic compression experiments. The results show that the displacement change of the electrode hook hair after 50 compression cycles was still less than 1%. This electrode provides a universal acquisition scheme, including effective acquisition of different parts of the body with or without hair. Finally, the gesture recognition from electromyography (EMG) by the CVE electrode was applied with accuracy above 90%. The CVE proposed in this study has great potential and promise in various human–machine interface (HMI) applications that employ surface biopotential signals on the body or head with hair.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186547","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mariella Särestöniemi, Daljeet Singh, Mikael von und zu Fraunberg, Teemu Myllylä
Microwave (MW) sensing is regarded as a promising technique for various medical monitoring and diagnostic applications due to its numerous advantages and the potential to be developed into a portable device for use outside hospital settings. The detection of skull fractures and the monitoring of their healing process would greatly benefit from a rapidly and frequently usable application that can be employed outside the hospital. This paper presents a simulation- and experiment-based study on skull fracture detection with the MW technique using realistic models for the first time. It also presents assessments on the most promising frequency ranges for skull fracture detection within the Industrial, Scientific and Medical (ISM) and ultrawideband (UWB) ranges. Evaluations are carried out with electromagnetic simulations using different head tissue layer models corresponding to different locations in the human head, as well as an anatomically realistic human head simulation model. The measurements are conducted with a real human skull combined with tissue phantoms developed in our laboratory. The comprehensive evaluations show that fractures cause clear differences in antenna and channel parameters (S11 and S21). The difference in S11 is 0.1–20 dB and in S21 is 0.1–30 dB, depending on the fracture width and location. Skull fractures with a less than 1 mm width can be detected with microwaves at different fracture locations. The detectability is frequency dependent. Power flow representations illustrate how fractures impact on the signal propagation at different frequencies. MW-based detection of skull fractures provides the possibility to (1) detect fractures using a safe and low-cost portable device, (2) monitor the healing-process of fractures, and (3) bring essential information for emerging portable MW-based diagnostic applications that can detect, e.g., strokes.
{"title":"Microwave Technique for Linear Skull Fracture Detection—Simulation and Experimental Study Using Realistic Human Head Models","authors":"Mariella Särestöniemi, Daljeet Singh, Mikael von und zu Fraunberg, Teemu Myllylä","doi":"10.3390/bios14090434","DOIUrl":"https://doi.org/10.3390/bios14090434","url":null,"abstract":"Microwave (MW) sensing is regarded as a promising technique for various medical monitoring and diagnostic applications due to its numerous advantages and the potential to be developed into a portable device for use outside hospital settings. The detection of skull fractures and the monitoring of their healing process would greatly benefit from a rapidly and frequently usable application that can be employed outside the hospital. This paper presents a simulation- and experiment-based study on skull fracture detection with the MW technique using realistic models for the first time. It also presents assessments on the most promising frequency ranges for skull fracture detection within the Industrial, Scientific and Medical (ISM) and ultrawideband (UWB) ranges. Evaluations are carried out with electromagnetic simulations using different head tissue layer models corresponding to different locations in the human head, as well as an anatomically realistic human head simulation model. The measurements are conducted with a real human skull combined with tissue phantoms developed in our laboratory. The comprehensive evaluations show that fractures cause clear differences in antenna and channel parameters (S11 and S21). The difference in S11 is 0.1–20 dB and in S21 is 0.1–30 dB, depending on the fracture width and location. Skull fractures with a less than 1 mm width can be detected with microwaves at different fracture locations. The detectability is frequency dependent. Power flow representations illustrate how fractures impact on the signal propagation at different frequencies. MW-based detection of skull fractures provides the possibility to (1) detect fractures using a safe and low-cost portable device, (2) monitor the healing-process of fractures, and (3) bring essential information for emerging portable MW-based diagnostic applications that can detect, e.g., strokes.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"80 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186548","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Katarína Nemčeková, Jana Korčeková, Veronika Svitková, Denis Baraniak, Michaela Domšicová, Eva Melníková, Michaela Hornychová, Viktória Szebellaiová, Miroslav Gál, Alexandra Poturnayová
The rapid and accurate detection of SARS-CoV-2, particularly its spike receptor-binding domain (S-RBD), was crucial for managing the COVID-19 pandemic. This study presents the development and optimization of two types of aptasensors: quartz crystal microbalance (QCM) and electrochemical sensors, both employing thiol-modified DNA aptamers for S-RBD detection. The QCM aptasensor demonstrated exceptional sensitivity, achieved by optimizing aptamer concentration, buffer composition, and pre-treatment conditions, with a limit of detection (LOD) of 0.07 pg/mL and a linear range from 1 pg/mL to 0.1 µg/mL, and a significant frequency change was observed upon target binding. The electrochemical aptasensor, designed for rapid and efficient preparation, utilized a one-step modification process that reduced the preparation time to 2 h while maintaining high sensitivity and specificity. Electrochemical impedance spectroscopy (EIS) enabled the detection of S-RBD concentrations as low as 132 ng/mL. Both sensors exhibited high specificity, with negligible non-specific interactions observed in the presence of competing proteins. Additionally, the QCM aptasensor’s functionality and stability were verified in biological fluids, indicating its potential for real-world applications. This study highlights the comparative advantages of QCM and electrochemical aptasensors in terms of preparation time, sensitivity, and specificity, offering valuable insights for the development of rapid, sensitive, and specific diagnostic tools for the detection of SARS-CoV-2 and other viruses.
{"title":"Comparative Analysis of QCM and Electrochemical Aptasensors for SARS-CoV-2 Detection","authors":"Katarína Nemčeková, Jana Korčeková, Veronika Svitková, Denis Baraniak, Michaela Domšicová, Eva Melníková, Michaela Hornychová, Viktória Szebellaiová, Miroslav Gál, Alexandra Poturnayová","doi":"10.3390/bios14090431","DOIUrl":"https://doi.org/10.3390/bios14090431","url":null,"abstract":"The rapid and accurate detection of SARS-CoV-2, particularly its spike receptor-binding domain (S-RBD), was crucial for managing the COVID-19 pandemic. This study presents the development and optimization of two types of aptasensors: quartz crystal microbalance (QCM) and electrochemical sensors, both employing thiol-modified DNA aptamers for S-RBD detection. The QCM aptasensor demonstrated exceptional sensitivity, achieved by optimizing aptamer concentration, buffer composition, and pre-treatment conditions, with a limit of detection (LOD) of 0.07 pg/mL and a linear range from 1 pg/mL to 0.1 µg/mL, and a significant frequency change was observed upon target binding. The electrochemical aptasensor, designed for rapid and efficient preparation, utilized a one-step modification process that reduced the preparation time to 2 h while maintaining high sensitivity and specificity. Electrochemical impedance spectroscopy (EIS) enabled the detection of S-RBD concentrations as low as 132 ng/mL. Both sensors exhibited high specificity, with negligible non-specific interactions observed in the presence of competing proteins. Additionally, the QCM aptasensor’s functionality and stability were verified in biological fluids, indicating its potential for real-world applications. This study highlights the comparative advantages of QCM and electrochemical aptasensors in terms of preparation time, sensitivity, and specificity, offering valuable insights for the development of rapid, sensitive, and specific diagnostic tools for the detection of SARS-CoV-2 and other viruses.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The liver has many important functions, including the biotransformation of drugs and detoxification of the human organism. As such, it is also exposed to many harmful substances, which leads to disorders and diseases such as cirrhosis. For these reasons, it seems important to consider liver metabolism and the direct effects on the liver when evaluating the efficacy of new drugs. Accordingly, we have developed an advanced in vitro liver model using an organ-on-a-chip approach that replicates many of the morphological and functional features of the liver in vivo. The model we created can metabolize drugs, which we demonstrated using two widely used anticancer drugs, 5-fluorouracil (5FU) and capecitabine (CAP). In addition, to the best of our knowledge, we are the first who evaluate the direct effects of these drugs not only on the viability of liver model-building cells but on their functions, such as cytochrome P450 activity and albumin production. Our study brings new hope to properly evaluating drug efficacy at the in vitro level.
{"title":"Advanced Liver-on-a-Chip Model for Evaluating Drug Metabolism and Hepatotoxicity","authors":"Sonia Frojdenfal, Agnieszka Zuchowska","doi":"10.3390/bios14090435","DOIUrl":"https://doi.org/10.3390/bios14090435","url":null,"abstract":"The liver has many important functions, including the biotransformation of drugs and detoxification of the human organism. As such, it is also exposed to many harmful substances, which leads to disorders and diseases such as cirrhosis. For these reasons, it seems important to consider liver metabolism and the direct effects on the liver when evaluating the efficacy of new drugs. Accordingly, we have developed an advanced in vitro liver model using an organ-on-a-chip approach that replicates many of the morphological and functional features of the liver in vivo. The model we created can metabolize drugs, which we demonstrated using two widely used anticancer drugs, 5-fluorouracil (5FU) and capecitabine (CAP). In addition, to the best of our knowledge, we are the first who evaluate the direct effects of these drugs not only on the viability of liver model-building cells but on their functions, such as cytochrome P450 activity and albumin production. Our study brings new hope to properly evaluating drug efficacy at the in vitro level.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"5 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186549","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Recent advances in three-dimensional (3D) culturing and nanotechnology offer promising pathways to overcome the limitations of drug screening, particularly for tumors like neuroblastoma. In this study, we develop a high-throughput microfluidic chip that integrates a concentration gradient generator (CGG) with a 3D co-culture system, constructing the vascularized microenvironment in tumors by co-culturing neuroblastoma (SY5Y cell line) and human brain microvascular endothelial cells (HBMVECs) within a decellularized extracellular matrix (dECM) hydrogels. The automated platform enhances the simulation of the tumor microenvironment and allows for the precise control of the concentrations of nanomedicines, which is crucial for evaluating therapeutic efficacy. The findings demonstrate that the high-throughput platform can significantly accelerate drug discovery. It efficiently screens and analyzes drug interactions in a biologically relevant setting, potentially revolutionizing the drug screening process.
{"title":"Tumor Microenvironment Based on Extracellular Matrix Hydrogels for On-Chip Drug Screening","authors":"Xiaoyan Liu, Jinxiong Cheng, Yingcan Zhao","doi":"10.3390/bios14090429","DOIUrl":"https://doi.org/10.3390/bios14090429","url":null,"abstract":"Recent advances in three-dimensional (3D) culturing and nanotechnology offer promising pathways to overcome the limitations of drug screening, particularly for tumors like neuroblastoma. In this study, we develop a high-throughput microfluidic chip that integrates a concentration gradient generator (CGG) with a 3D co-culture system, constructing the vascularized microenvironment in tumors by co-culturing neuroblastoma (SY5Y cell line) and human brain microvascular endothelial cells (HBMVECs) within a decellularized extracellular matrix (dECM) hydrogels. The automated platform enhances the simulation of the tumor microenvironment and allows for the precise control of the concentrations of nanomedicines, which is crucial for evaluating therapeutic efficacy. The findings demonstrate that the high-throughput platform can significantly accelerate drug discovery. It efficiently screens and analyzes drug interactions in a biologically relevant setting, potentially revolutionizing the drug screening process.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"2022 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nidhi Subhashini, Yannick Kerler, Marcus M. Menger, Olga Böhm, Judith Witte, Christian Stadler, Alexander Griberman
This study re-introduces a protein-free rapid test method for nucleic acids on paper based lateral flow assays utilizing special multichannel nitrocellulose membranes and DNA-Gold conjugates, achieving significantly enhanced sensitivity, easier protocols, reduced time of detection, reduced costs of production and advanced multiplexing possibilities. A protein-free nucleic acid-based lateral flow assay (NALFA) with a limit of detection of 1 pmol of DNA is shown for the first time. The total production duration of such an assay was successfully reduced from the currently known several days to just a few hours. The simplification and acceleration of the protocol make the method more accessible and practical for various applications. The developed method supports multiplexing, enabling the simultaneous detection of up to six DNA targets. This multiplexing capability is a significant improvement over traditional line tests and offers more comprehensive diagnostic potential in a single assay. The approach significantly reduces the run time compared to traditional line tests, which enhances the efficiency of diagnostic procedures. The protein-free aspect of this assay minimizes the prevalent complications of cross-reactivity in immunoassays especially in cases of multiplexing. It is also demonstrated that the NALFA developed in this study is amplification-free and hence does not rely on specialized technicians, nor does it involve labour-intensive steps like DNA extraction and PCR processes. Overall, this study presents a robust, efficient, and highly sensitive platform for DNA or RNA detection, addressing several limitations of current methods documented in the literature. The advancements in sensitivity, cost reduction, production time, and multiplexing capabilities mark a substantial improvement, holding great potential for various applications in diagnostics, forensics, and molecular biology.
本研究利用特殊的多通道硝酸纤维素膜和 DNA-Gold 结合物，在纸基横向流动检测法上重新引入了一种不含蛋白质的核酸快速检测方法，显著提高了灵敏度，简化了操作步骤，缩短了检测时间，降低了生产成本，并实现了先进的多重检测。首次展示了一种不含蛋白质的基于核酸的横向流动检测（NALFA），其 DNA 检测限为 1 pmol。这种检测方法的总生产时间成功地从目前已知的数天缩短到几小时。方案的简化和加速使该方法在各种应用中更加方便实用。所开发的方法支持多重检测，可同时检测多达六个 DNA 靶标。与传统的线性检测相比，这种多重检测能力是一项重大改进，可在一次检测中提供更全面的诊断潜力。与传统的线路测试相比，该方法大大缩短了运行时间，提高了诊断程序的效率。该检测方法不含蛋白质，最大程度地减少了免疫测定中普遍存在的交叉反应并发症，尤其是在多重检测的情况下。研究还表明，本研究开发的 NALFA 无需扩增，因此不依赖专业技术人员，也不涉及 DNA 提取和 PCR 过程等劳动密集型步骤。总之，本研究为 DNA 或 RNA 检测提供了一个稳健、高效、高灵敏度的平台，解决了文献中记载的当前方法的几个局限性。在灵敏度、成本降低、生产时间和多路复用能力方面的进步标志着一个实质性的改进，在诊断、法医和分子生物学的各种应用中蕴藏着巨大的潜力。
{"title":"Enhancing Colorimetric Detection of Nucleic Acids on Nitrocellulose Membranes: Cutting-Edge Applications in Diagnostics and Forensics","authors":"Nidhi Subhashini, Yannick Kerler, Marcus M. Menger, Olga Böhm, Judith Witte, Christian Stadler, Alexander Griberman","doi":"10.3390/bios14090430","DOIUrl":"https://doi.org/10.3390/bios14090430","url":null,"abstract":"This study re-introduces a protein-free rapid test method for nucleic acids on paper based lateral flow assays utilizing special multichannel nitrocellulose membranes and DNA-Gold conjugates, achieving significantly enhanced sensitivity, easier protocols, reduced time of detection, reduced costs of production and advanced multiplexing possibilities. A protein-free nucleic acid-based lateral flow assay (NALFA) with a limit of detection of 1 pmol of DNA is shown for the first time. The total production duration of such an assay was successfully reduced from the currently known several days to just a few hours. The simplification and acceleration of the protocol make the method more accessible and practical for various applications. The developed method supports multiplexing, enabling the simultaneous detection of up to six DNA targets. This multiplexing capability is a significant improvement over traditional line tests and offers more comprehensive diagnostic potential in a single assay. The approach significantly reduces the run time compared to traditional line tests, which enhances the efficiency of diagnostic procedures. The protein-free aspect of this assay minimizes the prevalent complications of cross-reactivity in immunoassays especially in cases of multiplexing. It is also demonstrated that the NALFA developed in this study is amplification-free and hence does not rely on specialized technicians, nor does it involve labour-intensive steps like DNA extraction and PCR processes. Overall, this study presents a robust, efficient, and highly sensitive platform for DNA or RNA detection, addressing several limitations of current methods documented in the literature. The advancements in sensitivity, cost reduction, production time, and multiplexing capabilities mark a substantial improvement, holding great potential for various applications in diagnostics, forensics, and molecular biology.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"33 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186553","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Uliana S. Novoyatlova, Anna A. Kudryavtseva, Sergey V. Bazhenov, Anna A. Utkina, Vadim V. Fomin, Shamil A. Nevmyanov, Bagila S. Zhoshibekova, Maria A. Fedyaeva, Mikhail Y. Kolobov, Ilya V. Manukhov
The ability of aquatic mesofauna representatives involved in trophic chains to sorb and accumulate toxicants is important for understanding the functioning of aquatic ecosystems and for fishing industry. This study investigated the capacity of marine amphipod Gammarus oceanicus and freshwater amphipods Eulimnogammarus vittatus and Gammarus lacustris to absorb the DNA-alkylating agent methyl methanesulfonate (MMS). The presence of alkylating agents in the environment and in the tissues of the amphipods was determined using whole-cell lux-biosensor Escherichia coli MG1655 pAlkA-lux, in which the luxCDABE genes from Photorhabdus luminescens, enabling the luminescence of the cell culture, are controlled by the PalkA promoter of DNA glycosylase. It was shown that within one day of incubation in water containing MMS at a concentration above 10 μM, the amphipods absorbed the toxicant and their tissues produce more alkylation damage to biosensor cells than the surrounding water. Concentrations of MMS above 1 mM in the environment caused the death of the amphipods before the toxicant could be significantly concentrated in their tissues. The sensitivity and the capacity to absorb MMS were found to be approximately the same for the marine amphipod G. oceanicus and the freshwater amphipods E. vittatus and G. lacustris.
参与营养链的水生中层动物吸附和积累有毒物质的能力对于了解水生生态系统的功能和渔业都很重要。本研究调查了海洋两栖动物大洋(Gammarus oceanicus)以及淡水两栖动物Eulimnogammarus vittatus和湖沼(Gammarus lacustris)吸收DNA烷化剂甲基磺酸盐(MMS)的能力。利用大肠杆菌 MG1655 pAlkA-lux 全细胞勒克斯生物传感器测定了环境和片脚类动物组织中存在的烷化剂。研究表明，在含有浓度超过 10 μM 的 MMS 的水中培养一天后，片脚类动物就会吸收毒性物质，其组织对生物传感器细胞产生的烷基化损伤比周围的水更大。环境中的 MMS 浓度超过 1 mM 会导致片脚类动物在毒物在其组织中显著富集之前死亡。研究发现，海洋片脚类动物 G. oceanicus 与淡水片脚类动物 E. vittatus 和 G. lacustris 对 MMS 的灵敏度和吸收能力大致相同。
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Mitochondria, crucial intracellular organelles, are central to energy metabolism, signal transduction, apoptosis, calcium homeostasis, and a myriad of other biological processes, making them a focal point in diverse research fields. The capacity to fluorescently label and visually track mitochondria is crucial for understanding their biological roles. We present mulberrin-Cy3, a novel small molecule fluorescent probe that selectively labels mitochondria in animal cells, including cancer cells, with relative ease. This protocol details the synthesis of mulberrin-Cy3 and its use for visualizing mitochondria in living cells. The synthesis is straightforward and time-efficient, and the labeling method is more accessible than traditional approaches, providing a cost-effective option for mitochondrial visualization at room temperature. The labeling is rapid, with effective labeling achieved within 5 min of incubation. The fluorescent signal is stable and brighter, offering a significant advantage over existing methods. Mulberrin-Cy3 represents a promising mitochondrial labeling compound, providing researchers with a novel experimental tool to explore the complex biological functions of mitochondria. This innovation has the potential to significantly advance our comprehension of mitochondrial dynamics and their role in cellular health and disease.
{"title":"Mitochondrial Labeling with Mulberrin-Cy3: A New Fluorescent Probe for Live Cell Visualization","authors":"Gangxiang Yuan, Yiwei Luo, Peng Qian, Ningjia He","doi":"10.3390/bios14090428","DOIUrl":"https://doi.org/10.3390/bios14090428","url":null,"abstract":"Mitochondria, crucial intracellular organelles, are central to energy metabolism, signal transduction, apoptosis, calcium homeostasis, and a myriad of other biological processes, making them a focal point in diverse research fields. The capacity to fluorescently label and visually track mitochondria is crucial for understanding their biological roles. We present mulberrin-Cy3, a novel small molecule fluorescent probe that selectively labels mitochondria in animal cells, including cancer cells, with relative ease. This protocol details the synthesis of mulberrin-Cy3 and its use for visualizing mitochondria in living cells. The synthesis is straightforward and time-efficient, and the labeling method is more accessible than traditional approaches, providing a cost-effective option for mitochondrial visualization at room temperature. The labeling is rapid, with effective labeling achieved within 5 min of incubation. The fluorescent signal is stable and brighter, offering a significant advantage over existing methods. Mulberrin-Cy3 represents a promising mitochondrial labeling compound, providing researchers with a novel experimental tool to explore the complex biological functions of mitochondria. This innovation has the potential to significantly advance our comprehension of mitochondrial dynamics and their role in cellular health and disease.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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