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Microfluidic Barcode Biochips for High-Throughput Real-Time Biomolecule and Single-Cell Screening 认领 引用 被引量：1
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作者 Jiaoyan Qiu Yanbo Liang +4 位作者 Chao Wang Yang Yu Yu Zhang Hong Liu Lin Han 《Engineering》 SCIE EI CSCD 2025年第3期130-146,共17页
The real-time screening of biomolecules and single cells in biochips is extremely important for disease prediction and diagnosis,cellular analysis,and life science research.Barcode biochip technology,which is integrat... The real-time screening of biomolecules and single cells in biochips is extremely important for disease prediction and diagnosis,cellular analysis,and life science research.Barcode biochip technology,which is integrated with microfluidics,typically comprises barcode array,sample loading,and reaction unit array chips.Here,we present a review of microfluidics barcode biochip analytical approaches for the high-throughput screening of biomolecules and single cells,including protein biomarkers,microRNA(miRNA),circulating tumor DNA(ctDNA),single-cell secreted proteins,single-cell exosomes,and cell interactions.We begin with an overview of current high-throughput detection and analysis approaches.Following this,we outline recent improvements in microfluidic devices for biomolecule and single-cell detection,highlighting the benefits and limitations of these devices.This paper focuses on the research and development of microfluidic barcode biochips,covering their self-assembly substrate materials and their specific applications with biomolecules and single cells.Looking forward,we explore the prospects and challenges of this technology,with the aim of contributing toward the use of microfluidic barcode detection biochips in medical diagnostics and therapies,and their large-scale commercialization. 展开更多
关键词 High-throughput Microfluidic barcode biochip Single-cell analysis Biomolecules
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Single-cell deoxyribonucleic acid typing for forensic mixtures and trace evidence:Opportunities,validation requirements,and reporting limits 认领 引用
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作者 Kyriacos Evangelou Paraskevi Angeli +1 位作者 Andreas Polydorou Thalia Petropoulou 《World Journal of Biological Chemistry》 2026年第2期1-9,共9页
Forensic deoxyribonucleic acid(DNA)interpretation is limited less by genotyping technology than by the biochemical and inferential effects of mixed,low-template,and environmentally complex traces.Single-cell and singl... Forensic deoxyribonucleic acid(DNA)interpretation is limited less by genotyping technology than by the biochemical and inferential effects of mixed,low-template,and environmentally complex traces.Single-cell and single-molecule strategies,including the United Kingdom Research and Innovation-funded single-cell and single-molecule analysis for DNA identification(SCAnDi)program,aim to preserve cellular resolution.They allow investigators to isolate and type individual cells or defined small-cell pools before heterogeneous evidence is converted into a bulk lysate.In selected validation settings,this approach has yielded near-complete diploid short tandem repeat(STR)profiles from small pools,credible genotype sets tightly concentrated on the true genotype across high-order mixtures,and improved access to donor-specific profiles from sexual assault and other complex samples.However,these studies also show important limits.Many operationally successful“single-cell”workflows are,in practice,single-cell-plusconsensus or few-cell workflows.Stochastic effects remain intrinsic,and cell capture itself becomes a probabilistic sampling step.This opinion review explicitly adopts an evaluative stance:It synthesizes recent validation studies and guidance documents to identify the performance thresholds and reporting boundaries that should be met before targeted forensic deployment of SCAnDilike workflows.We argue that casework entry should require measured cell-recovery probabilities,phenotypemisclassification rates,locus-and cell-type-specific dropout and stutter models,quantified contamination and drop-in rates,validated minimum cell counts for consensus generation,and explicit database-upload criteria.It should also require strict separation between sub-source reporting and activity-level propositions.If those conditions are met,single-cell typing can complement,rather than replace,bulk STR analysis and probabilistic genotyping in a narrow but important set of high-value forensic scenarios. 展开更多
关键词 Forensic genetics Single-cell genomics DNA mixtures Laser capture microdissection Digital cell sorting Whole-genome amplification Short tandem repeats Activity-level propositions DNA transfer
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An integrated microfluidics platform with high-throughput single-cell cloning array and concentration gradient generator for efficient cancer drug effect screening 认领 引用 被引量：3
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作者 Biao Wang Bang-Shun He +6 位作者 Xiao-Lan Ruan Jiang Zhu Rui Hu Jie Wang Ying Li Yun-Huang Yang Mai-Li Liu 《Military Medical Research》 SCIE CAS CSCD 2023年第3期325-341,共17页
Background:Tumor cell heterogeneity mediated drug resistance has been recognized as the stumbling block of cancer treatment.Elucidating the cytotoxicity of anticancer drugs at single-cell level in a high-throughput wa... Background:Tumor cell heterogeneity mediated drug resistance has been recognized as the stumbling block of cancer treatment.Elucidating the cytotoxicity of anticancer drugs at single-cell level in a high-throughput way is thus of great value for developing precision therapy.However,current techniques suffer from limitations in dynamically characterizing the responses of thousands of single cells or cell clones presented to multiple drug conditions.Methods:We developed a new microfluidics-based“SMART”platform that is Simple to operate,able to generate a Massive single-cell array and Multiplex drug concentrations,capable of keeping cells Alive,Retainable and Trackable in the microchambers.These features are achieved by integrating a Microfluidic chamber Array(4320 units)and a sixConcentration gradient generator(MAC),which enables highly efficient analysis of leukemia drug effects on single cells and cell clones in a high-throughput way.Results:A simple procedure produces 6 on-chip drug gradients to treat more than 3000 single cells or single-cell derived clones and thus allows an efficient and precise analysis of cell heterogeneity.The statistic results reveal that Imatinib(Ima)and Resveratrol(Res)combination treatment on single cells or clones is much more efficient than Ima or Res single drug treatment,indicated by the markedly reduced half maximal inhibitory concentration(IC50).Additionally,single-cell derived clones demonstrate a higher IC50 in each drug treatment compared to single cells.Moreover,primary cells isolated from two leukemia patients are also found with apparent heterogeneity upon drug treatment on MAC.Conclusions:This microfluidics-based“SMART”platform allows high-throughput single-cell capture and culture,dynamic drug-gradient treatment and cell response monitoring,which represents a new approach to efficiently investigate anticancer drug effects and should benefit drug discovery for leukemia and other cancers. 展开更多
关键词 Microfluidics Single-cell analysis Leukemia High-throughput drug screening Single-cell cloning
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High-throughput sorting of two-color fluorescent-labeled zebrafish embryos 认领 引用
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作者 Hongzhen Tang Linbo Wang +3 位作者 Xiaohu Chen Chong Chen Hui Li Guang Yang 《Journal of Innovative Optical Health Sciences》 SCIE EI CSCD 2023年第5期107-114,共8页
The zebrafish embryos were widely employed in genetics,development and drug discovery studies as miniatured animal models.Sorting of two-color fluorescent embryos is often required in large-scale experiments but it is... The zebrafish embryos were widely employed in genetics,development and drug discovery studies as miniatured animal models.Sorting of two-color fluorescent embryos is often required in large-scale experiments but it is challenging to manually sort with high efficiency.Here,we reported a high-throughput sorting system for two-color fluorescent zebraflsh embryos.The embryos can be automatically loaded from a sample pool and sorted based on the average fluorescent intensity.The two-color fluorescent signals were split into two lines and detected by an area array camera.The system achieves the sorting of 100 embryos in less than 10 min with an accuracy of greater than 95%. 展开更多
关键词 High-throughput zebrafish embryo two-color sorting automatic.
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A time-coded multi-concentration microfluidic chemical waveform generator for high-throughput probing suspension single-cell signaling 认领 引用
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作者 Yiran Guo Peng Chen +4 位作者 Zhaolong Gao Yiwei Li Shunji Li Xiaojun Feng Bi-Feng Liu 《Chinese Chemical Letters》 SCIE CAS CSCD 2022年第6期3091-3096,共6页
The cellular response to the complex extracellular microenvironment is highly dynamic in time and type of extracellular matrix.Accurately reconstructing this process and analyzing the changes in receptor conformation ... The cellular response to the complex extracellular microenvironment is highly dynamic in time and type of extracellular matrix.Accurately reconstructing this process and analyzing the changes in receptor conformation on the cell membrane surface and intracellular or intercellular signaling has been a major challenge in analytical chemistry and biophysical methodology.In this paper,a time-coded multiconcentration microfluidic chemical waveform generator was developed for the dynamic signaling probing with single-cell array of high temporal resolution,high throughput,and multi-concentration combination stimulation.Based on innovative microchannel structure,sophisticated external control methods and multiplexing technology,the system not only allowed for temporally sequential permutations of the four concentrations of stimuli(time code),but also generated pulsed and continuous waveforms at different frequencies in a highly controllable manner.Furthermore,the single-cell trap array was set up to efficiently capture cells in suspension,dramatically increasing throughput and reducing experiment preparation time.The maximum frequency of the platform was 1 Hz,and one cell could be stimulated at multiple frequencies.To show the ability of the system to investigate rapid biochemical events in high throughput,pulse stimulation and continuous stimulation of different frequencies and different time codes,combined with four concentrations of histamine(HA),were generated for probing G protein-coupled receptor(GPCR)signaling in He La cells.Then,statistical analysis was performed for the mean peak height and mean peak area of the cellular response.We believe that the time-coded multi-concentration microfluidic chemical waveform generator will provide a novel strategy for analytical chemistry,biophysics,cell signaling,and individualized medicine applications. 展开更多
关键词 Microfluidics Dynamic stimulation Time code High-throughput Single-cell array
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Artificial intelligence-assisted automatic and index-based microbial single-cell sorting system for One-Cell-One-Tube 认领 引用 被引量：7
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作者 Zhidian Diao Lingyan Kan +11 位作者 Yilong Zhao Jingyun Song Chen Wang Yang Liu Fengli Zhang Teng Xu Rongze Chen Xixian Wang Xiaoyan Jing Jian Xu Yuandong Li Bo Ma 《mLife》 2022年第4期448-459,共12页
Identification,sorting,and sequencing of individual cells directly from in situ samples have great potential for in-depth analysis of the structure and function of microbiomes.In this work,based on an artificial intel... Identification,sorting,and sequencing of individual cells directly from in situ samples have great potential for in-depth analysis of the structure and function of microbiomes.In this work,based on an artificial intelligence(AI)-assisted object detection model for cell phenotype screening and a cross-interface contact method for single-cell exporting,we developed an automatic and index-based system called EasySort AUTO,where individual microbial cells are sorted and then packaged in a microdroplet and automatically exported in a precisely indexed,“One-Cell-One-Tube”manner.The target cell is automatically identified based on an AI-assisted object detection model and then mobilized via an optical tweezer for sorting.Then,a crossinterface contact microfluidic printing method that we developed enables the automated transfer of cells from the chip to the tube,which leads to coupling with subsequent single-cell culture or sequencing.The efficiency of the system for single-cell printing is>93%.The throughput of the system for single-cell printing is~120 cells/h.Moreover,>80%of single cells of both yeast and Escherichia coli are culturable,suggesting the superior preservation of cell viability during sorting.Finally,AI-assisted object detection supports automated sorting of target cells with high accuracy from mixed yeast samples,which was validated by downstream single-cell proliferation assays.The automation,index maintenance,and vitality preservation of EasySort AUTO suggest its excellent application potential for single-cell sorting. 展开更多
关键词 artificial intelligence One-Cell-One-Tube(OCOT) single-cell analysis single-cell printing single-cell sorting
Validation of breast cancer as a risk factor for anxiety and depression:Insights from Mendelian randomization analysis 认领 引用
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作者 Guannan He Man Xi +2 位作者 Tianhao Zhang Shuang Wang Gang Liu 《Journal of Pharmaceutical Analysis》 SCIE CAS CSCD 2025年第9期2101-2121,共21页
This study employed Mendelian randomization(MR)analysis to confirm the association between breast cancer and the risk of anxiety and depression,and to explore the molecular mechanisms by which lipid nanoparticles of k... This study employed Mendelian randomization(MR)analysis to confirm the association between breast cancer and the risk of anxiety and depression,and to explore the molecular mechanisms by which lipid nanoparticles of ketamine(LNP@Ket)modulate these behaviors in a mouse model of breast cancer.Through single-cell transcriptomic analysis,the study aimed to clarify nuclear factor erythroid 2-related factor 2(Nrf2)’s role in the development of anxiety and depression in these mice.Analysis of patient data from genome-wide association study(GWAS)databases supported the link between breast cancer,anxiety,and depression.In vivo experiments demonstrated that treating breast cancer mice with LNP@Ket significantly reduced anxiety and depression behaviors.The synthesis of LNP@Ket and its subsequent analysis highlighted its inhibitory effects on these behaviors.Single-cell transcriptomic sequencing identified key cells and genes affected by LNP@Ket treatment,particularly emphasizing Nrf2.Upregulation of Nrf2 in astrocytes increased the expression of antioxidant enzymes and reduced pro-inflammatory cytokines,alleviating anxiety and depression symptoms by inhibiting neuroinflammation and neurodegeneration.This comprehensive study highlights the pivotal role of Nrf2 in the therapeutic efficacy of LNP@Ket for treating anxiety and depression in breast cancer mice. 展开更多
关键词 Breast cancer Anxiety and depression behaviors Ketamine-loaded lipid nanoparticlesNrf2 AstrocytesMendelian randomization analysis Single-cell transcriptome sequencing High-throughput transcriptome sequencing
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Genetically encoded biosensors enabled high-throughput screening of microbial cell factories 认领 引用
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作者 Jin Wang Xueyan Liu +2 位作者 Longqian Zhao Yue Zhang Meng Wang 《Engineering Microbiology》 CSCD 2026年第1期85-97,共13页
Genetically encoded biosensors provide powerful tools for coupling desired phenotypes to detectable outputs and have been extensively developed to detect a wide range of natural and unnatural products.When integrated ... Genetically encoded biosensors provide powerful tools for coupling desired phenotypes to detectable outputs and have been extensively developed to detect a wide range of natural and unnatural products.When integrated with diverse high-throughput screening(HTS)approaches,these biosensors enable efficient product-driven screening across various throughputs,thereby expediting the engineering and optimization of microbial cell factories to produce various target compounds.For effective HTS of microbial cell factories,biosensors need to possess certain crucial characteristics.The performance features of biosensors significantly influence their application potential in HTS and can be precisely engineered through synthetic biology strategies.Furthermore,to ensure biosensordriven HTS,additional engineering and optimizations of the biosensors are often required to increase the success rate and reduce false positives in the screening process.This review discusses the essential features of genetically encoded biosensors designed for HTS and then summarizes the latest advances in biosensor engineering for HTS purposes via synthetic biology strategies.Following this,the challenges and optimization of biosensors to adapt to different HTS processes are also discussed and exemplified.Finally,the key concerns and research prospects of developing biosensors for HTS applications are highlighted.Overall,this review provides comprehensive guidance on the engineering of genetically encoded biosensors and their applications in HTS for developing microbial cell factories to produce diverse target compounds. 展开更多
关键词 Genetically encoded biosensor High-throughput screening Microbial cell factory Fluorescence-activated cell sorting Fluorescence-activated droplet sorting Synthetic biology Artificial intelligence
Recent advances in high-throughput droplet-based microfluidic screening platforms for food analysis 认领 引用 被引量：1
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作者 Qianqian Zhang Shiqiang Yue +3 位作者 Dongyuan Zhang Fuping Lu Shuhong Mao Hui-Min Qin 《Food Bioscience》 SCIE 2024年第6期1192-1203,共12页
As a novel type of miniaturized instrumentation,high-throughput droplet-based microfluidics is playing an increasingly significant role in food safety inspection and quality control.The popularization of traditional i... As a novel type of miniaturized instrumentation,high-throughput droplet-based microfluidics is playing an increasingly significant role in food safety inspection and quality control.The popularization of traditional instrumental analysis is largely limited by the need for lengthy analysis time and costly instrumentation.The novel high-throughput droplet-based microfluidic screening platform(DMSP)has advantages of high screening rate,single-cell packaging,and less reagent consumption that solves the drawbacks of traditional instruments.In this review,we introduce the generation and manipulation of high-throughput DMSP,after which we summarize their recent applications in food analysis.These applications were demonstrated in the directed evolution of microorganisms for fermented food,improving the catalytic efficiency of enzymes for food processing,screening of nutraceutical ingredients,and detection of hazardous substances.We also provide a critical evaluation of the state of high-throughput DMSP and suggestions for future development directions. 展开更多
关键词 Droplet-based microfluidic screening platform High-throughput screening Single-cell packaging Food safety inspection Food quality control
The application of microfluidic-based technologies in the cycle of metabolic engineering 认领 引用 被引量：4
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作者 Xiaoyan Ma Yi-Xin Huo 《Synthetic and Systems Biotechnology》 SCIE 2016年第3期137-142,共6页
The process of metabolic engineering consists of multiple cycles of design,build,test and learn,which is typically laborious and time-consuming.To increase the efficiency and the rate of success of strain engineering,... The process of metabolic engineering consists of multiple cycles of design,build,test and learn,which is typically laborious and time-consuming.To increase the efficiency and the rate of success of strain engineering,novel instrumentation must be applied.Microfluidics,the control of liquid flow in microstructures,has enabled flexible,accurate,automatic,and high-throughput manipulation of cells in liquid at picoliter to nanoliter scale.These attributes hold great promise in advancing metabolic engineering in terms of the phases of design,build,test and learn.To promote the application of microfluidic-based technologies in strain improvement,this review addressed the potentials of microfluidics and the related approaches in DNA assembly,transformation,strain screening,genotyping and phenotyping,and highlighted their adaptations for single-cell analysis.As a result,this facilitates in-depth understanding of the metabolic network,which in turn promote efficient optimization in the following cycles of strain engineering.Taken together,microfluidic-based technologies enable on-chip workflow,and could greatly accelerate the turnaround of metabolic engineering. 展开更多
关键词 Metabolic engineering Microfluidics High-throughput Single-cell analysis
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