Miaomiao Xu, Siyuan Wei, Tong Su, Die Ma, Zhixuan Wang, Dan Zhu, Lixing Weng, Xianguang Ding
Macrophages, known for their phenotypic plasticity, play a critical role in maintaining homeostasis and inflammation-related pathogenesis. Although identifying diverse macrophage phenotypes holds promise for enhancing diagnoses and treatments of diseases mediated by macrophages, existing methodologies for differentiating macrophages often lack precision. They are limited by the cumbersome procedures that require large-scale equipment, such as flow cytometry and transcriptomic analysis. In this context, we have engineered fluorescent polyadenine (polyA)-mediated sticky flares that enable practical visualization of macrophages. This technology facilitates the highly sensitive detection of macrophage phenotypes through the specific recognition of intracellular mRNAs, permitting in situ imaging. Our approach demonstrates the potential for determining macrophage polarization status at the single-cell level within dynamic immune microenvironments, thereby providing crucial diagnostic and prognostic information that could guide the development of tailored treatments for macrophage-related diseases in personalized medicine.
{"title":"Visualizing Macrophage Polarization through Fluorescent mRNA Profiling.","authors":"Miaomiao Xu, Siyuan Wei, Tong Su, Die Ma, Zhixuan Wang, Dan Zhu, Lixing Weng, Xianguang Ding","doi":"10.3390/bios14100475","DOIUrl":"https://doi.org/10.3390/bios14100475","url":null,"abstract":"<p><p>Macrophages, known for their phenotypic plasticity, play a critical role in maintaining homeostasis and inflammation-related pathogenesis. Although identifying diverse macrophage phenotypes holds promise for enhancing diagnoses and treatments of diseases mediated by macrophages, existing methodologies for differentiating macrophages often lack precision. They are limited by the cumbersome procedures that require large-scale equipment, such as flow cytometry and transcriptomic analysis. In this context, we have engineered fluorescent polyadenine (polyA)-mediated sticky flares that enable practical visualization of macrophages. This technology facilitates the highly sensitive detection of macrophage phenotypes through the specific recognition of intracellular mRNAs, permitting in situ imaging. Our approach demonstrates the potential for determining macrophage polarization status at the single-cell level within dynamic immune microenvironments, thereby providing crucial diagnostic and prognostic information that could guide the development of tailored treatments for macrophage-related diseases in personalized medicine.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11506351/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510886","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Detecting circulating tumor cells has exhibited great significance in treating cancers since its concentration is an index strongly associated with the development and transfer of the tumor. However, the present commercial method for CTC detection is still expensive, because special antibodies and complicated devices must be used for cell separation and imaging. Hence, it is quite necessary to apply alternative materials and methods to decrease the cost of CTC detection. In this article, we coated a cellulose acetate membrane with nanoparticles formed by the polymerization of melamine and furfural, creating a surface with nanoscale roughness for the highly efficient capture of the sparse CTCs in a blood sample. Subsequently, the CTCs on the surface can be quantitatively detected by colorimetry with the aid of a COF-based nanozyme. The detection limit (LOD) can be as low as 3 cells/mL, which is the lowest LOD among the colorimetric methods to our knowledge. Considering the low cost of fabricating the membrane for CTC capture and the robustness of nanozymes compared with natural enzymes, this CTC detection approach displays great potential to decrease the financial burden of commercial CTC detection.
{"title":"A Nanoparticle-Coated Cellulose Acetate Membrane for Highly Efficient, Low-Cost Circulating Tumor Cell Detection.","authors":"Yize Zhao, Yaqi Pan, Hao Sun, Pengfei Huo, Guangtong Wang, Shaoqin Liu","doi":"10.3390/bios14100472","DOIUrl":"https://doi.org/10.3390/bios14100472","url":null,"abstract":"<p><p>Detecting circulating tumor cells has exhibited great significance in treating cancers since its concentration is an index strongly associated with the development and transfer of the tumor. However, the present commercial method for CTC detection is still expensive, because special antibodies and complicated devices must be used for cell separation and imaging. Hence, it is quite necessary to apply alternative materials and methods to decrease the cost of CTC detection. In this article, we coated a cellulose acetate membrane with nanoparticles formed by the polymerization of melamine and furfural, creating a surface with nanoscale roughness for the highly efficient capture of the sparse CTCs in a blood sample. Subsequently, the CTCs on the surface can be quantitatively detected by colorimetry with the aid of a COF-based nanozyme. The detection limit (LOD) can be as low as 3 cells/mL, which is the lowest LOD among the colorimetric methods to our knowledge. Considering the low cost of fabricating the membrane for CTC capture and the robustness of nanozymes compared with natural enzymes, this CTC detection approach displays great potential to decrease the financial burden of commercial CTC detection.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11505997/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Microfluidics are crucial for managing small-volume analytical solutions for various applications, such as disease diagnostics, drug efficacy testing, chemical analysis, and water quality monitoring. The precise control of flow control devices can generate diverse flow patterns using pneumatic control with solenoid valves and a microcontroller. This system enables the active modulation of the pneumatic pressure through Arduino programming of the solenoid valves connected to the pressure source. Additionally, the incorporation of solenoid valve sets allows for multichannel control, enabling simultaneous creation and manipulation of various microflows at a low cost. The proposed microfluidic flow controller facilitates accurate flow regulation, especially through periodic flow modulation beneficial for droplet generation and continuous production of microdroplets of different sizes. Overall, we expect the proposed microfluidic flow controller to drive innovative advancements in technology and medicine owing to its engineering precision and versatility.
{"title":"Microscale Flow Control and Droplet Generation Using Arduino-Based Pneumatically-Controlled Microfluidic Device.","authors":"Woohyun Park, Se-Woon Choe, Minseok Kim","doi":"10.3390/bios14100469","DOIUrl":"https://doi.org/10.3390/bios14100469","url":null,"abstract":"<p><p>Microfluidics are crucial for managing small-volume analytical solutions for various applications, such as disease diagnostics, drug efficacy testing, chemical analysis, and water quality monitoring. The precise control of flow control devices can generate diverse flow patterns using pneumatic control with solenoid valves and a microcontroller. This system enables the active modulation of the pneumatic pressure through Arduino programming of the solenoid valves connected to the pressure source. Additionally, the incorporation of solenoid valve sets allows for multichannel control, enabling simultaneous creation and manipulation of various microflows at a low cost. The proposed microfluidic flow controller facilitates accurate flow regulation, especially through periodic flow modulation beneficial for droplet generation and continuous production of microdroplets of different sizes. Overall, we expect the proposed microfluidic flow controller to drive innovative advancements in technology and medicine owing to its engineering precision and versatility.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11506217/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Minimally invasive surgery continues to prioritize patient safety by improving imaging techniques and tumor detection methods. In this work, an all-optical alternative to the current image based techniques for in vitro minimally invasive procedures has been explored. The technique uses a highly fluorescent marker for the surgical needle to be tracked inside simulated tissues. A series of markers were explored including inorganic (Perovskite and PbS) and organic (carbon dots) nanoparticles and organic dye (Rhodamine 6G) to identify layers of different stiffnesses within a tissue. Rhodamine 6G was chosen based on its high fluorescence signal to track 3D position of a surgical needle in a tissue. The needle was tracked inside homogeneous and inhomogeneous gelatin tissues successfully. This exploratory study of tissue characterization and needle tip tracking using fluorescent markers or photoluminescence technique show potential for real-time application of robot-assisted needle insertions during in vivo procedures.
{"title":"Needle Tip Tracking through Photoluminescence for Minimally Invasive Surgery.","authors":"Meenakshi Narayan, Mithun Bhowmick","doi":"10.3390/bios14100470","DOIUrl":"https://doi.org/10.3390/bios14100470","url":null,"abstract":"<p><p>Minimally invasive surgery continues to prioritize patient safety by improving imaging techniques and tumor detection methods. In this work, an all-optical alternative to the current image based techniques for in vitro minimally invasive procedures has been explored. The technique uses a highly fluorescent marker for the surgical needle to be tracked inside simulated tissues. A series of markers were explored including inorganic (Perovskite and PbS) and organic (carbon dots) nanoparticles and organic dye (Rhodamine 6G) to identify layers of different stiffnesses within a tissue. Rhodamine 6G was chosen based on its high fluorescence signal to track 3D position of a surgical needle in a tissue. The needle was tracked inside homogeneous and inhomogeneous gelatin tissues successfully. This exploratory study of tissue characterization and needle tip tracking using fluorescent markers or photoluminescence technique show potential for real-time application of robot-assisted needle insertions during in vivo procedures.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11505679/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510844","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ryan H P Siu, Robert G Jesky, Yu-Jing Fan, Cyrus C H Au-Yeung, Andrew B Kinghorn, Kwok-Hung Chan, Ivan Fan-Ngai Hung, Julian A Tanner
Gold standard detection of SARS-CoV-2 by reverse transcription quantitative PCR (RT-qPCR) can achieve ultrasensitive viral detection down to a few RNA copies per sample. Yet, the lengthy detection and labor-intensive protocol limit its effectiveness in community screening. In view of this, a structural switching electrochemical aptamer-based biosensor (E-AB) targeting the SARS-CoV-2 nucleocapsid (N) protein was developed. Four N protein-targeting aptamers were characterized on an electrochemical cell configuration using square wave voltammetry (SWV). The sensor was investigated in an artificial saliva matrix optimizing the aptamer anchoring orientation, SWV interrogation frequency, and target incubation time. Rapid detection of the N protein was achieved within 5 min at a low nanomolar limit of detection (LOD) with high specificity. Specific N protein detection was also achieved in simulated positive saliva samples, demonstrating its feasibility for saliva-based rapid diagnosis. Further research will incorporate novel signal amplification strategies to improve sensitivity for early diagnosis.
通过反转录定量 PCR(RT-qPCR)检测 SARS-CoV-2 的金标准可实现超灵敏病毒检测,每个样本可检测到几个 RNA 拷贝。然而,冗长的检测过程和劳动密集型方案限制了其在社区筛查中的有效性。有鉴于此,我们开发了一种以 SARS-CoV-2 核头蛋白(N)为靶标的结构转换电化学适配体生物传感器(E-AB)。利用方波伏安法(SWV)在电化学电池配置上对四种 N 蛋白靶向适配体进行了表征。在人工唾液基质中对传感器进行了研究,优化了适配体的锚定方向、SWV 检测频率和目标培养时间。在 5 分钟内就实现了对 N 蛋白的快速检测,检测限(LOD)低至纳摩尔,特异性高。在模拟的阳性唾液样本中也实现了对 N 蛋白的特异性检测,证明了其在基于唾液的快速诊断中的可行性。进一步的研究将采用新型信号放大策略,以提高早期诊断的灵敏度。
{"title":"Aptamer-Mediated Electrochemical Detection of SARS-CoV-2 Nucleocapsid Protein in Saliva.","authors":"Ryan H P Siu, Robert G Jesky, Yu-Jing Fan, Cyrus C H Au-Yeung, Andrew B Kinghorn, Kwok-Hung Chan, Ivan Fan-Ngai Hung, Julian A Tanner","doi":"10.3390/bios14100471","DOIUrl":"https://doi.org/10.3390/bios14100471","url":null,"abstract":"<p><p>Gold standard detection of SARS-CoV-2 by reverse transcription quantitative PCR (RT-qPCR) can achieve ultrasensitive viral detection down to a few RNA copies per sample. Yet, the lengthy detection and labor-intensive protocol limit its effectiveness in community screening. In view of this, a structural switching electrochemical aptamer-based biosensor (E-AB) targeting the SARS-CoV-2 nucleocapsid (N) protein was developed. Four N protein-targeting aptamers were characterized on an electrochemical cell configuration using square wave voltammetry (SWV). The sensor was investigated in an artificial saliva matrix optimizing the aptamer anchoring orientation, SWV interrogation frequency, and target incubation time. Rapid detection of the N protein was achieved within 5 min at a low nanomolar limit of detection (LOD) with high specificity. Specific N protein detection was also achieved in simulated positive saliva samples, demonstrating its feasibility for saliva-based rapid diagnosis. Further research will incorporate novel signal amplification strategies to improve sensitivity for early diagnosis.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11505747/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The evaluation of the upkeep and freshness of aquatic products within the cold chain is crucial due to their perishable nature, which can significantly impact both quality and safety. Conventional methods for assessing freshness in the cold chain have inherent limitations regarding specificity and accuracy, often requiring substantial time and effort. Recently, advanced sensor technologies have been developed for freshness assessment, enabling real-time and non-invasive monitoring via the detection of volatile organic compounds, biochemical markers, and physical properties. The integration of sensor technologies into cold chain logistics enhances the ability to maintain the quality and safety of aquatic products. This review examines the advancements made in multifunctional sensor devices for the freshness assessment of aquatic products in cold chain logistics, as well as the application of pattern recognition algorithms for identification and classification. It begins by outlining the categories of freshness criteria, followed by an exploration of the development of four key sensor devices: electronic noses, electronic tongues, biosensors, and flexible sensors. Furthermore, the review discusses the implementation of advanced pattern recognition algorithms in sensor devices for freshness detection and evaluation. It highlights the current status and future potential of sensor technologies for aquatic products within the cold chain, while also addressing the significant challenges that remain to be overcome.
{"title":"A Review of Advanced Sensor Technologies for Aquatic Products Freshness Assessment in Cold Chain Logistics.","authors":"Baichuan Wang, Kang Liu, Guangfen Wei, Aixiang He, Weifu Kong, Xiaoshuan Zhang","doi":"10.3390/bios14100468","DOIUrl":"https://doi.org/10.3390/bios14100468","url":null,"abstract":"<p><p>The evaluation of the upkeep and freshness of aquatic products within the cold chain is crucial due to their perishable nature, which can significantly impact both quality and safety. Conventional methods for assessing freshness in the cold chain have inherent limitations regarding specificity and accuracy, often requiring substantial time and effort. Recently, advanced sensor technologies have been developed for freshness assessment, enabling real-time and non-invasive monitoring via the detection of volatile organic compounds, biochemical markers, and physical properties. The integration of sensor technologies into cold chain logistics enhances the ability to maintain the quality and safety of aquatic products. This review examines the advancements made in multifunctional sensor devices for the freshness assessment of aquatic products in cold chain logistics, as well as the application of pattern recognition algorithms for identification and classification. It begins by outlining the categories of freshness criteria, followed by an exploration of the development of four key sensor devices: electronic noses, electronic tongues, biosensors, and flexible sensors. Furthermore, the review discusses the implementation of advanced pattern recognition algorithms in sensor devices for freshness detection and evaluation. It highlights the current status and future potential of sensor technologies for aquatic products within the cold chain, while also addressing the significant challenges that remain to be overcome.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11506179/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510864","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Particle separation and sorting techniques based on microfluidics have found extensive applications and are increasingly gaining prominence. This research presents the design and fabrication of a microfluidic device for separating cells using deterministic lateral displacement (DLD), enabling accuracy and continuity while being size-based. Nevertheless, it remains demanding, to completely reverse the detrimental effects of the boundaries that disturb the fluidic flow in the channel and reduce particle separation efficiency. This study introduces a novel approach to enhance the boundary structure of channels. By using this design, separation efficiency is boosted, and the fluid behavior around the walls is improved. The boundary correction (BC) enhances the operation of the microchannel and is very effective in microchannels. With boundary correction, the device exhibited improved separation efficiencies, but in its absence, separation efficiencies dropped. The collected microscopic images of the isolation of prostate cancer cell lines and red blood cells revealed promising outcomes. The efficiency of circulating tumor cell (CTC) throughput in the microfluidic channel, quantified as the ratio or proportion of tumor cells exiting the channel to cells entering it, exceeds 93%. Moreover, the efficiency of CTC isolation, expressed as the proportion of tumor cells from the upper outlet of the microfluidic channel to all cells, is over 89%. Additionally, the efficiency of red blood cell isolation, evaluated as the ratio of red blood cells from the lower outlet of the microfluidic channel to all cells, surpasses 77%. While using the same DLD separator without boundary correction reduced the separation efficiency by around 5%.
{"title":"Effective Boundary Correction for Deterministic Lateral Displacement Microchannels to Improve Cell Separation: A Numerical and Experimental Study.","authors":"Shaghayegh Mirhosseini, Mohammadmahdi Eskandarisani, Aryanaz Faghih Nasiri, Fatemeh Khatami, Akram Mirzaei, Majid Badieirostami, Seyed Mohammad Kazem Aghamir, Mohammadreza Kolahdouz","doi":"10.3390/bios14100466","DOIUrl":"https://doi.org/10.3390/bios14100466","url":null,"abstract":"<p><p>Particle separation and sorting techniques based on microfluidics have found extensive applications and are increasingly gaining prominence. This research presents the design and fabrication of a microfluidic device for separating cells using deterministic lateral displacement (DLD), enabling accuracy and continuity while being size-based. Nevertheless, it remains demanding, to completely reverse the detrimental effects of the boundaries that disturb the fluidic flow in the channel and reduce particle separation efficiency. This study introduces a novel approach to enhance the boundary structure of channels. By using this design, separation efficiency is boosted, and the fluid behavior around the walls is improved. The boundary correction (BC) enhances the operation of the microchannel and is very effective in microchannels. With boundary correction, the device exhibited improved separation efficiencies, but in its absence, separation efficiencies dropped. The collected microscopic images of the isolation of prostate cancer cell lines and red blood cells revealed promising outcomes. The efficiency of circulating tumor cell (CTC) throughput in the microfluidic channel, quantified as the ratio or proportion of tumor cells exiting the channel to cells entering it, exceeds 93%. Moreover, the efficiency of CTC isolation, expressed as the proportion of tumor cells from the upper outlet of the microfluidic channel to all cells, is over 89%. Additionally, the efficiency of red blood cell isolation, evaluated as the ratio of red blood cells from the lower outlet of the microfluidic channel to all cells, surpasses 77%. While using the same DLD separator without boundary correction reduced the separation efficiency by around 5%.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11506467/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510806","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cutaneous squamous cell carcinoma (cSCC) is the second most common malignant skin tumor. Early and precise diagnosis of tumor staging is crucial for long-term outcomes. While pathological diagnosis has traditionally served as the gold standard, the assessment of differentiation levels heavily depends on subjective judgments. Therefore, how to improve the diagnosis accuracy and objectivity of pathologists has become an urgent problem to be solved. We used multispectral imaging (MSI) to enhance tumor classification. The hematoxylin and eosin (H&E) stained cSCC slides were from Shanghai Ruijin Hospital. Scale-invariant feature transform was applied to multispectral images for image stitching, while the adaptive threshold segmentation method and random forest segmentation method were used for image segmentation, respectively. Synthetic pseudo-color images effectively highlight tissue differences. Quantitative analysis confirms significant variation in the nuclear area between normal and cSCC tissues (p < 0.001), supported by an AUC of 1 in ROC analysis. The AUC within cSCC tissues is 0.57. Further study shows higher nuclear atypia in poorly differentiated cSCC tissues compared to well-differentiated cSCC (p < 0.001), also with an AUC of 1. Lastly, well differentiated cSCC tissues show more and larger keratin pearls. These results have shown that combined MSI with imaging processing techniques will improve H&E stained human cSCC diagnosis accuracy, and it will be well utilized to distinguish histopathological staging features.
{"title":"Distinguishing of Histopathological Staging Features of H-E Stained Human cSCC by Microscopical Multispectral Imaging.","authors":"Rujuan Wu, Jiayi Yang, Qi Chen, Changxing Yang, Qianqian Ge, Danni Rui, Huazhong Xiang, Dawei Zhang, Cheng Wang, Xiaoqing Zhao","doi":"10.3390/bios14100467","DOIUrl":"https://doi.org/10.3390/bios14100467","url":null,"abstract":"<p><p>Cutaneous squamous cell carcinoma (cSCC) is the second most common malignant skin tumor. Early and precise diagnosis of tumor staging is crucial for long-term outcomes. While pathological diagnosis has traditionally served as the gold standard, the assessment of differentiation levels heavily depends on subjective judgments. Therefore, how to improve the diagnosis accuracy and objectivity of pathologists has become an urgent problem to be solved. We used multispectral imaging (MSI) to enhance tumor classification. The hematoxylin and eosin (H&E) stained cSCC slides were from Shanghai Ruijin Hospital. Scale-invariant feature transform was applied to multispectral images for image stitching, while the adaptive threshold segmentation method and random forest segmentation method were used for image segmentation, respectively. Synthetic pseudo-color images effectively highlight tissue differences. Quantitative analysis confirms significant variation in the nuclear area between normal and cSCC tissues (<i>p</i> < 0.001), supported by an AUC of 1 in ROC analysis. The AUC within cSCC tissues is 0.57. Further study shows higher nuclear atypia in poorly differentiated cSCC tissues compared to well-differentiated cSCC (<i>p</i> < 0.001), also with an AUC of 1. Lastly, well differentiated cSCC tissues show more and larger keratin pearls. These results have shown that combined MSI with imaging processing techniques will improve H&E stained human cSCC diagnosis accuracy, and it will be well utilized to distinguish histopathological staging features.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11506349/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Single nucleotide variant (SNV) detection is pivotal in various fields, including disease diagnosis, viral screening, genetically modified organism (GMO) identification, and genotyping. However, detecting SNVs presents significant challenges due to the fragmentation of nucleic acids caused by cellular apoptosis, molecular shearing, and physical degradation processes such as heating. Fragmented nucleic acids often exhibit variable lengths and inconsistent breakpoints, complicating the accurate detection of SNVs. This article delves into the underlying causes of nucleic acid fragmentation and synthesizes the strengths and limitations of next-generation sequencing technology, high-resolution melting curves, molecular probes, and CRISPR-based approaches for SNV detection in fragmented nucleic acids. By providing a detailed comparative analysis, it seeks to offer valuable insights for researchers working to overcome the challenges of SNV detection in fragmented samples, ultimately advancing the accurate and efficient detection of single nucleotide variants across diverse applications.
{"title":"Advances in Genotyping Detection of Fragmented Nucleic Acids.","authors":"Qian Liu, Yun Chen, Hao Qi","doi":"10.3390/bios14100465","DOIUrl":"https://doi.org/10.3390/bios14100465","url":null,"abstract":"<p><p>Single nucleotide variant (SNV) detection is pivotal in various fields, including disease diagnosis, viral screening, genetically modified organism (GMO) identification, and genotyping. However, detecting SNVs presents significant challenges due to the fragmentation of nucleic acids caused by cellular apoptosis, molecular shearing, and physical degradation processes such as heating. Fragmented nucleic acids often exhibit variable lengths and inconsistent breakpoints, complicating the accurate detection of SNVs. This article delves into the underlying causes of nucleic acid fragmentation and synthesizes the strengths and limitations of next-generation sequencing technology, high-resolution melting curves, molecular probes, and CRISPR-based approaches for SNV detection in fragmented nucleic acids. By providing a detailed comparative analysis, it seeks to offer valuable insights for researchers working to overcome the challenges of SNV detection in fragmented samples, ultimately advancing the accurate and efficient detection of single nucleotide variants across diverse applications.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11506436/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The urine albumin (Alb)-to-creatinine (Crn) ratio (UACR) is a sensitive and early indicator of chronic kidney disease (CKD) and cardiorenal syndrome. This study developed a portable and wireless electrochemical-sensing platform for the sensitive and accurate determination of UACR. The developed platform consists of a carbon nanotube (CNT)-2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)(ABTS)-based modified UACR sensor, a miniaturised potentiostat, a cup holder embedded with a magnetic stirrer and a smartphone app. The UACR sensing electrode is composed of two screen-printed carbon working electrodes, one screen-printed carbon counter electrode and a screen-printed AgCl reference electrode. The miniaturised potentiostat, which is controlled by the developed app, performs cyclic voltammetry and amperometry to detect Alb and Crn, respectively. Clinical trials of the proposed system by using spot urine samples from 30 diabetic patients indicate that it can accurately classify all three CKD risk statuses within 30 min. The high accuracy of our proposed sensing system exhibits satisfactory agreement with the commercial biochemical analyser TBA-25FR (Y = 0.999X, R2 = 0.995). The proposed UACR sensing system offers a convenient, reliable and affordable solution for personal mobile health monitoring and point-of-care urinalysis.
{"title":"Portable Electrochemical System and Platform with Point-of-Care Determination of Urine Albumin-to-Creatinine Ratio to Evaluate Chronic Kidney Disease and Cardiorenal Syndrome.","authors":"Shuenn-Yuh Lee, Ding-Siang Ciou, Hao-Yun Lee, Ju-Yi Chen, Yi-Chieh Wei, Meng-Dar Shieh","doi":"10.3390/bios14100463","DOIUrl":"https://doi.org/10.3390/bios14100463","url":null,"abstract":"<p><p><b></b> The urine albumin (Alb)-to-creatinine (Crn) ratio (UACR) is a sensitive and early indicator of chronic kidney disease (CKD) and cardiorenal syndrome. This study developed a portable and wireless electrochemical-sensing platform for the sensitive and accurate determination of UACR. The developed platform consists of a carbon nanotube (CNT)-2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)(ABTS)-based modified UACR sensor, a miniaturised potentiostat, a cup holder embedded with a magnetic stirrer and a smartphone app. The UACR sensing electrode is composed of two screen-printed carbon working electrodes, one screen-printed carbon counter electrode and a screen-printed AgCl reference electrode. The miniaturised potentiostat, which is controlled by the developed app, performs cyclic voltammetry and amperometry to detect Alb and Crn, respectively. Clinical trials of the proposed system by using spot urine samples from 30 diabetic patients indicate that it can accurately classify all three CKD risk statuses within 30 min. The high accuracy of our proposed sensing system exhibits satisfactory agreement with the commercial biochemical analyser TBA-25FR (Y = 0.999X, R<sup>2</sup> = 0.995). The proposed UACR sensing system offers a convenient, reliable and affordable solution for personal mobile health monitoring and point-of-care urinalysis.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11506532/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510850","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}