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Enhancing single-cell encapsulation in droplet microfluidics with fine-tunable on-chip sample enrichment

Microsystems & Nanoengineering 2024年第1期10卷 149-160页

作者： Tao Tang Hao Zhao Shaofei Shen Like Yang Chwee Teck Lim Department of Biomedical engineering National University of Singapore117583 SingaporeSingapore integrative sciences and engineering Programme NUS Graduate SchoolNational University of Singapore119077 SingaporeSingapore Shanxi Key Lab for Modernization of TCVM College of Life ScienceShanxi Agricultural UniversityTaiguShanxi 030801China Institute for Health Innovation&Technology National University of Singapore117599 SingaporeSingapore Mechanobiology Institute National University of Singapore117411 SingaporeSingapore Institute for Digital Molecular Analytics and Science Nanyang Technological University636921 SingaporeSingapore

Single-cell encapsulation in droplet microfluidics is commonly hindered by the tradeoff between cell suspension density and on-chip focusing performance.In this study,we introduce a novel droplet microfluidic chip to overcome this challenge.The chip comprises a double spiral focusing unit,a flow resistance-based sample enrichment module with fine-tunable outlets,and a crossflow droplet generation unit.Utilizing a low-density cell/bead suspension(2×10^(6) objects/mL),cells/beads are focused into a near-equidistant linear arrangement within the double spiral microchannel.The excess water phase is diverted while cells/beads remain focused and sequentially encapsulated in individual droplets.Focusing performance was assessed through numerical simulations and experiments at three flow rates(40,60,80μL/min),demonstrating successful focusing at 40 and 80μL/min for beads and cells,respectively.In addition,both simulation and experimental results revealed that the flow resistance at the sample enrichment module is adjustable by punching different outlets,allowing over 50%of the aqueous phase to be removed.YOLOv8n-based droplet detection algorithms realized the counting of cells/beads in droplets,statistically demonstrating single-cell and bead encapsulation rates of 72.2%and 79.2%,respectively.All the results indicate that this on-chip sample enrichment approach can be further developed and employed as a critical component in single-cell encapsulation in water-in-oil droplets.

关键词： beads outlet sample

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Chinese herbal medicine Ginkgo biloba L.preparations for ischemic stroke:An overview of systematic reviews and meta-analyses

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Journal of integrative Medicine 2024年第2期22卷 163-179页

作者： Tian-tian Meng Ya-ping You Min Li Jian-bo Guo Xin-bin Song Jing-yi Ding Xiao-long Xie An-qi Li Shang-jin Li Xiang-jun Yin Peng Wang Zhe Wang Bao-liang Wang Qing-yong He Department of Cardiology Guang'anmen HospitalChina Academy of Chinese Medical SciencesBeijing 100032China Department of Rehabilitation Dongfang HospitalBeijing University of Chinese MedicineBeijing 100071China school of Chemistry and Chemical engineering Shanghai Jiao Tong UniversityShanghai 200240China school of Chinese Medicine the University of Hong KongHong KongChina graduate school Henan University of Chinese MedicineZhengzhou 450008Henan ProvinceChina Department of Acupuncture and Moxibustion Dongzhimen HospitalBeijing University of Chinese MedicineBeijing 100010China Department of Cardiology Guang’anmen HospitalChina Academy of Chinese Medical SciencesZhengzhou 450008Henan ProvinceChina school of Integrated Traditional Chinese and Western Medicine Southwest Medical UniversityHangzhou 310053Zhejiang ProvinceChina school of Basic Medical sciences Zhejiang Chinese Medical UniversityHangzhu 310053Zhejang ProvinceChina Department of Neurology the First Affiliated Hospital of Henan University of Chinese MedicineZhengzhou 450003Henan ProvinceChina

Background:Ginkgo biloba L.preparations(GBLPs)are a class of Chinese herbal medicine used in the adjuvant treatment of ischemic stroke(IS).Recently,several systematic reviews(SRs)and meta-analyses(MAs)of GBLPs for IS have been published.Objective:This overview aims to assess the quality of related SRs and MAs.Search strategy:Pub Med,Embase,Cochrane Library,Web of Science,Chinese Biological Medicine,China National Knowledge Infrastructure,Wanfang,and Chinese Science and Technology Journals databases were searched from their inception to December 31,2022.Inclusion criteria:SRs and MAs of randomized controlled trials(RCTs)that explored the efficacy of GBLPs for patients with IS were included.Data extraction and analysis:Two independent reviewers extracted data and assessed the methodological quality,risk of bias(ROB),reporting quality,and credibility of evidence of the included SRs and MAs using A Measurement Tool to Assess Systematic Reviews 2(AMSTAR 2),Risk of Bias in Systematic Reviews(ROBIS),the Preferred Reporting Items for Systematic Reviews and Meta-analyses(PRISMA),and the Grading of Recommendations Assessment,Development and Evaluation(GRADE),respectively.Additionally,descriptive analysis and data synthesis were conducted.Results:Twenty-nine SRs/MAs involving 119 outcomes were included in this review.The overall methodological quality of all SRs/MAs was critically low based on AMSTAR 2,and 28 had a high ROB based on the ROBIS.According to the PRISMA statement,the reporting items of the included SRs/MAs are relatively complete.The results based on GRADE showed that of the 119 outcomes,8 were rated as moderate quality,24 as low quality,and 87 as very low quality.Based on the data synthesis,GBLPs used in conjunction with conventional treatment were superior to conventional treatment alone for decreasing neurological function scores.Conclusion:GBLPs can be considered a beneficial supplemental therapy for IS.However,because of the low quality of the existing evidence,high-quality RCTs and SRs/MAs are warranted t

关键词： statement synthesis alone

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Upconversion Nanoparticles-Encoded Hydrogel Microbeads-Based Multiplexed Protein Detection

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Nano-Micro Letters 2018年第2期10卷 135-150页

作者： Swati Shikha Xiang Zheng Yong Zhang Department of Biomedical engineering Faculty of Engineering National University of Singapore (NUS) nus graduate school for integrative sciences and engineering Centre for Life Sciences (CeLS)

Fluorescently encoded microbeads are in demand for multiplexed applications in different fields.Compared to organic dye-based commercially available Luminex's x MAP technology, upconversion nanoparticles(UCNPs) are better alternatives due to their large antiStokes shift, photostability, nil background, and single wavelength excitation. Here, we developed a new multiplexed detection system using UCNPs for encoding poly(ethylene glycol) diacrylate(PEGDA) microbeads as well as for labeling reporter antibody. However, to prepare UCNPs-encoded microbeads, currently used swellingbased encapsulation leads to non-uniformity, which is undesirable for fluorescence-based multiplexing. Hence,we utilized droplet microfluidics to obtain encoded microbeads of uniform size, shape, and UCNPs distribution inside. Additionally, PEGDA microbeads lack functionality for probe antibodies conjugation on their surface.Methods to functionalize the surface of PEGDA microbeads(acrylic acid incorporation, polydopamine coating)reported thus far quench the fluorescence of UCNPs. Here,PEGDA microbeads surface was coated with silica followed by carboxyl modification without compromising the fluorescence intensity of UCNPs. In this study, droplet microfluidics-assisted UCNPs-encoded microbeads of uniform shape, size, and fluorescence were prepared.Multiple color codes were generated by mixing UCNPs emitting red and green colors at different ratios prior to encapsulation. UCNPs emitting blue color were used to label the reporter antibody. Probe antibodies were covalently immobilized on red UCNPs-encoded microbeads for specific capture of human serum albumin(HSA) as a model protein. The system was also demonstrated for multiplexed detection of both human C-reactive protein(hCRP) and HSA protein by immobilizing anti-h CRP antibodies on green UCNPs.

关键词： Upconversion nanoparticles PEGDA microbeads Encoding Multiplexed bio-detection Single wavelength excitation

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3D Printing of Next-generation Electrochemical Energy Storage Devices: from Multiscale to Multimaterial

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Energy & Environmental Materials 2022年第2期5卷 427-438页

作者： Xi Xu Yong Hao Tan Jun Ding Cao Guan Institute of Flexible Electronics Xi’an Key Laboratory of Flexible ElectronicsNorthwestern Polytechnical UniversityXi’an 710072China Department of Materials Science and engineering Faculty of EngineeringNational University of SingaporeSingapore 117574Singapore nus graduate school for integrative sciences and engineering National University of SingaporeSingapore 119077Singapore

The increasing energy requirements to power the modern world has driven active research into more advanced electrochemical energy storage devices(EESD)with both high energy densities and power densities.Wide range of newly discovered materials with promising electrochemical properties has shown great potential for next-generation devices,but their performance is normally associated with contradicting demands of thin electrodes and high mass loading that can be hardly achieved for practical applications.Design of three-dimensional(3D)porous electrodes can increase the mass loading while maintaining the effective charge transport even with thick electrodes,which has proven to be efficient to overcome the limitations.3D structures have also been demonstrated excellent structural stability to withstand strong strains and stresses generated during charge/discharge cycle.3D printing,which can fabricate various delicate and complex structural designs,thus offering brand-new opportunities for the rational design and facile construction of next-generation EESDs.The recent developments in 3D printing of next-generation EESDs with high performance are reviewed.Advanced/multiscale electrode structures,such as hierarchically porous structure that can be constructed via high-resolution 3D printing or with post-treatment,are further emphasized.The ability of current 3D printing techniques to fulfill multimaterial printing to fulfill simple packaging will be covered.

关键词： 3d printing electrochemical energy storage devices multimaterial multiscale

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A Review on Deterministic Lateral Displacement for Particle Separation and Detection

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Nano-Micro Letters 2019年第4期11卷 353-385页

作者： Thoriq Salafi Yi Zhang Yong Zhang nus graduate school for integrative sciences and engineering National University of SingaporeSingapore 119077Singapore Department of Biomedical engineering National University of SingaporeSingapore 117583Singapore

The separation and detection of particles in suspension are essential for a wide spectrum of applications including medical diagnostics.In this field,microfluidic deterministic lateral displacement(DLD)holds a promise due to the ability of continuous separation of particles by size,shape,deformability,and electrical properties with high resolution.DLD is a passive microfluidic separation technique that has been widely implemented for various bioparticle separations from blood cells to exosomes.DLD techniques have been previously reviewed in 2014.Since then,the field has matured as several physics of DLD have been updated,new phenomena have been discovered,and various designs have been presented to achieve a higher separation performance and throughput.Furthermore,some recent progress has shown new clinical applications and ability to use the DLD arrays as a platform for biomolecules detection.This review provides a thorough discussion on the recent progress in DLD with the topics based on the fundamental studies on DLD models and applications for particle separation and detection.Furthermore,current challenges and potential solutions of DLD are also discussed.We believe that a comprehensive understanding on DLD techniques could significantly contribute toward the advancements in the field for various applications.In particular,the rapid,low-cost,and high-throughput particle separation and detection with DLD have a tremendous impact for point-of-care diagnostics.

关键词： Microfluidic Deterministic lateral displacement Particle separation Particle detection

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