1,852 results on '"DROPLET MICROFLUIDICS"'
Search Results
2. Microfluidic synthesis of stimuli-responsive hydrogel particles
- Author
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Zhao, Danshan, Qian, Lu, Yang, Qiaoyi, Li, Xiang, Ye, Chao, Shi, Tianqiong, and Wang, Yuetong
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- 2025
- Full Text
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3. Cell driven elastomeric particle packing in composite bioinks for engineering and implantation of stable 3D printed structures
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Landau, Shira, Kieda, Jennifer, Khosravi, Ramak, Okhovatian, Sargol, Ramsay, Kaitlyn, Liu, Chuan, Shakeri, Amid, Zhao, Yimu, Shen, Karen, Bar-Am, Orit, Levenberg, Shulamit, Tsai, Scott, and Radisic, Milica
- Published
- 2025
- Full Text
- View/download PDF
4. Flexible droplet microfluidic devices for tuneable droplet generation
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Roshan, Uditha, Dai, Yuchen, Yadav, Ajeet Singh, Hettiarachchi, Samith, Mudugamuwa, Amith, Zhang, Jun, and Nguyen, Nam-Trung
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- 2025
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5. Droplet-based single-cell sequencing: Strategies and applications
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Shang, Yuting, Wang, Zhengzheng, Xi, Liqing, Wang, Yantao, Liu, Meijing, Feng, Ying, Wang, Juan, Wu, Qingping, Xiang, Xinran, Chen, Moutong, and Ding, Yu
- Published
- 2024
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- View/download PDF
6. Machine learning-integrated droplet microfluidic system for accurate quantification and classification of microplastics
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Jeon, Ji Woo, Choi, Ji Wook, Shin, Yonghee, Kang, Taewook, and Chung, Bong Geun
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- 2025
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- View/download PDF
7. Single-cell sorting using integrated pneumatic valve droplet microfluidic chip
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Zhou, Yang, Yu, Zhibin, Wu, Man, Lan, Yuwei, Jia, Chunping, and Zhao, Jianlong
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- 2023
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8. Application of Flow Cytometry in the Development of New Alternative Model Systems in Biomedical Research
- Author
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Pandey, Anuj Kumar, Kulshrestha, Manish Raj, Singh, Mukul, Pathak, Anumesh K., Tiwari, Vandana, Pant, AB, editor, Khare, Puneet, editor, and Pandey, Alok Kumar, editor
- Published
- 2025
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9. Polymeric Microspheres with High Mass Fraction of Therapeutics Enabled by the Manipulation of Kinetics Factor During Emulsion Droplet Solidification.
- Author
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Wei, Zhenyang, Zhu, Mingyu, Morin, Nicolas, Wollsten, Daniela, Hirvonen, Jouni, Yang, Xiangliang, Santos, Hélder A., and Li, Wei
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CRYSTALLINE polymers , *MANIPULATION therapy , *DRUG interactions , *SOLIDIFICATION , *MICROSPHERES , *POLYCAPROLACTONE - Abstract
High drug‐loaded polymeric microspheres hold promise in biomedical fields due to reduced excipient administration, minimized side effects, and enhanced therapeutical efficacy. Although thermodynamic factors like drug‐carrier material compatibility are well‐known to influence the drug loading capacity of microspheres, they fail to explain the huge difference in drug loading degree observed for polymers and drugs with similar interactions. Here, based on the droplet microfluidic platform, the single droplet solidification process is investigated. The results indicated that amorphous polymers can hinder drug diffusion during droplet solidification compared to crystalline polymers, resulting in a higher drug loading degree. Next, this principle is applied to improve the drug loading capability of crystalline polymers (polycaprolactone (PCL) and poly(L‐lactide) (PLLA)) by random co‐polymerization (poly(caprolactone‐co‐L‐lactide) (PCL‐PLLA)), achieving 6.2–22.2 times increased drug loading degree. Moreover, PCL‐PLLA microspheres with a high content of indomethacin exhibited superior therapeutical efficacy in the treatment of gout arthritis. Overall, these results offer insights into the impact of polymer crystallization on droplet solidification kinetics, which consequently affects the drug loading capacity. These findings provide guidelines for the development of polymers for efficient drug encapsulation. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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10. Multimodal Imaging, Drug Delivery, and On‐Board Triggered Degradation in Soft Capsule Rolling Microrobots.
- Author
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Castellanos‐Robles, David, Doineau, Raphaël C. L.‐M., Aziz, Azaam, Nauber, Richard, Wu, Song, Moreno, Silvia, Mitropoulou, Konstantina, Hebenstreit, Franziska, and Medina‐Sánchez, Mariana
- Subjects
ULTRASOUND contrast media ,OPTICAL control ,CONTRAST media ,DRUG carriers ,MICROROBOTS - Abstract
In the rapidly advancing field of medical microrobotics, designing robots capable of addressing various challenges—such as imaging, biodegradation, and multifunctionality—is crucial. Departing from conventional research that often focuses on isolated aspects of microrobot functionality, this study presents an innovative approach to comprehensive microrobot design. Soft capsule microrobots that integrate capabilities such as magnetic navigation, autonomous maneuverability, in situ biodegradation, biosafe imaging, and drug delivery are reported. These microrobots are fabricated within the range of 20–120 μm, with a notable throughput of ≈102–103 microrobots per second. Furthermore, their locomotion performance has been demonstrated to remain stable for a period exceeding 10 h, all while employing real‐time optical closed‐loop control. The incorporation of ultrasound contrast agents not only amplifies imaging resolution but also ensures imaging contrast stability in a biological environment for over a period of 3 h. Second, the intentional integration of enzyme‐loaded nanometric polymersomes establishes a self‐contained, biodegradable system, accentuating the microrobots' capacity to degrade without the addition of high enzyme concentrations. This integrated approach lays the groundwork for minimally invasive treatments toward personalized and targeted medicine. [ABSTRACT FROM AUTHOR]
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- 2024
- Full Text
- View/download PDF
11. Artificial intelligence-based droplet size prediction for microfluidic system
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Dubey, Sameer, Vishwakarma, Pradeep, Ramarao, TVS, Dubey, Satish Kumar, Goel, Sanket, and Javed, Arshad
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- 2024
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12. A Microfluidic Multiplex Sorter for Strain Development.
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Leal‐Alves, Chiara, Dumont, Sebastien, Deng, Zhiyang, Alkhaldi, Sarah, Leung, Ziuwin, Oeser, Michelle, and Shih, Steve C. C.
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HIGH throughput screening (Drug development) , *BIOLOGICAL fitness , *GLUCOAMYLASE , *SORTING devices , *MICROFLUIDICS - Abstract
Selecting strains with superior traits from strain improvement strategies is challenging, as it involves navigating the fitness landscape by applying selective pressures that drive variants from peaks of improvement to valleys over time. In recent years, the screening and selection is conducted via droplet microfluidic methods due to its high throughput capabilities. However, the oft‐used binary strategy, targeting only the high levels of improved traits, may not reflect the overall enhancement. A multiplexed sorting method capable of applying an additional threshold to sort traits by phenotypic strength is reported. The novel approach uses a droplet‐digital microfluidic sorter to screen different volumes of droplets using the same device design and sorting parameters. This method is used to sort glucoamylase enzyme mutants with two levels of activity (medium and high) from libraries of diastatic yeast that have been mutated with non‐genetically modified techniques. Using the multiplex system, medium‐performing strains with enhanced (up to 60%) fermentation kinetics in synthetic beverage media, which would have been missed with a binary screening approach, are identified. The multiplex sorting strategy efficiently finds strains with superior fermentation traits in the fitness landscape without requiring extensive screening rounds and mutations. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
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13. Microfluidic Fabrication and Applications of Microgels and Hybrid Microgels.
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Farooqi, Zahoor H., Vladisavljević, Goran T., Pamme, Nicole, Fatima, Arooj, Begum, Robina, Irfan, Ahmad, and Chen, Minjun
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MICROGELS , *MOLECULAR weights , *MICROFLUIDICS , *GELATION , *MONOMERS , *MONODISPERSE colloids - Abstract
Smart microgels have gained much attention because of their wide range of applications in the field of biomedical, environmental, nanotechnological and catalysis sciences. Most of the applications of microgels are strongly affected by their morphology, size and size distribution. Various methodologies have been adopted to obtain polymer microgel particles. Droplet microfluidic techniques have been widely reported for the fabrication of highly monodisperse microgel particles to be used for various applications. Monodisperse microgel particles of required size and morphology can be achieved via droplet microfluidic techniques by simple polymerization of monomers in the presence of suitable crosslinker or by gelation of high molecular weight polymers. This report gives recent research progress in fabrication, characterization, properties and applications of microgel particles synthesized by microfluidic methods. [ABSTRACT FROM AUTHOR]
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- 2024
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14. A Simple Pump-Free Approach to Generating High-Throughput Microdroplets Using Oscillating Microcone Arrays.
- Author
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Yetiskin, Erturan, Erdem, Ilayda, Gucluer, Sinan, and Ozcelik, Adem
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SHEET metal ,MICROFLUIDICS ,MICRODROPLETS ,INDUSTRIAL applications ,SCALABILITY - Abstract
Droplet generation is crucial in various scientific and industrial fields, such as drug delivery, diagnostics, and inkjet printing. While microfluidic platforms enable precise droplet formation, traditional methods often require costly and complex setups, limiting their accessibility. This study introduces a simple, low-cost approach using an off-the-shelf unit and a 3D-printed reservoir. The device, equipped with a driver board, piezo-ring transducer, and a metal sheet with holes, generates oil-in-water (O/W) droplets with an average diameter of 4.62 ± 0.67 µm without external fluid pumps. Its simplicity, cost-effectiveness, and scalability make it highly suitable for both lab-on-chip and industrial applications, demonstrating the feasibility of large-scale uniform droplet production. [ABSTRACT FROM AUTHOR]
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- 2024
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15. Synthesis of Anisotropic Gold Microparticles via L‐Glutathione‐Mediated Pathways in Droplet Microfluidics.
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Yang, Zhenxu, Yin, Qiankun, He, Mengfan, Chong, Shin‐Wei, Xu, Zhejun, Liu, Xiaochen, Vega‐Sánchez, Christopher, Jaiswal, Arun, Vigolo, Daniele, and Yong, Ken‐Tye
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NANOPARTICLE synthesis , *FLOW chemistry , *MICROFLUIDICS , *NANOSTRUCTURED materials , *NANOPARTICLES , *GOLD nanoparticles - Abstract
Microfluidic‐assisted synthesis of nanoparticles has generated significant interest for its precise control and high throughput capabilities. Among various nanomaterials, gold nanoparticles (AuNPs) have shown remarkable potential in numerous applications, such as disease detection, photothermotherapy, drug delivery, and even defense applications. Recent synthesis strategy of peptide‐mediated method has sparked greater interest by offering unique chiroptical properties and their applications in biomedical applications. In this study, the use of droplet microfluidics is explored for the synthesis of peptide‐mediated AuNPs, aiming to accelerate automated production via flow chemistry. This method leads to the formation of anisotropic gold particles, with sizes ranging from hundreds of nanometers to the micron scale. The interfacial energy is identified at the water/oil interface as a critical factor influencing this outcome, with L‐glutathione (L‐GSH) playing a significant role in the development of hyper‐branched structures. These results demonstrate the capability of droplet microfluidics in producing anisotropic gold particles at micron scales, presenting new possibilities for the advancement of nanoparticle synthesis techniques. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
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16. High‐throughput G protein‐coupled receptor‐based autocrine screening for secondary metabolite production in yeast.
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Saleski, Tatyana E., Peng, Huadong, Lengger, Bettina, Wang, Jinglin, Jensen, Michael Krogh, and Jensen, Emil D.
- Abstract
Biosensors are valuable tools in accelerating the test phase of the design‐build‐test‐learn cycle of cell factory development, as well as in bioprocess monitoring and control. G protein‐coupled receptor (GPCR)‐based biosensors enable cells to sense a wide array of molecules and environmental conditions in a specific manner. Due to the extracellular nature of their sensing, GPCR‐based biosensors require compartmentalization of distinct genotypes when screening production levels of a strain library to ensure that detected levels originate exclusively from the strain under assessment. Here, we explore the integration of production and sensing modalities into a single Saccharomyces cerevisiae strain and compartmentalization using three different methods: (1) cultivation in microtiter plates, (2) spatial separation on agar plates, and (3) encapsulation in water‐in‐oil‐in‐water double emulsion droplets, combined with analysis and sorting via a fluorescence‐activated cell sorting machine. Employing tryptamine and serotonin as proof‐of‐concept target molecules, we optimize biosensing conditions and demonstrate the ability of the autocrine screening method to enrich for high producers, showing the enrichment of a serotonin‐producing strain over a nonproducing strain. These findings illustrate a workflow that can be adapted to screening for a wide range of complex chemistry at high throughput using commercially available microfluidic systems. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
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17. Fluorescence-activated droplet sequencing (FAD-seq) directly provides sequences of screening hits in antibody discovery.
- Author
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Autour, Alexis and Merten, Christoph A.
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HIGH throughput screening (Drug development) , *PHENOTYPES , *MICROFLUIDICS , *IMMUNOGLOBULINS , *REVERSE transcriptase polymerase chain reaction - Abstract
Droplet microfluidics has become a very powerful tool in high-throughput screening, including antibody discovery. Screens are usually carried out by physically sorting droplets hosting cells of the desired phenotype, breaking them, recovering the encapsulated cells, and sequencing the paired antibody light and heavy chain genes at the single-cell level. This series of multiple consecutive manipulation steps of rare screening hits is complex and challenging, resulting in a significant loss of clones with the desired phenotype or large fractions of cells with incomplete antibody information. Here, we present fluorescence-activated droplet sequencing, in which droplets showing the desired phenotype are selectively picoinjected with reagents for RT-PCR. Subsequently, light and heavy chain genes are natively paired, fused into a single-chain fragment variant format, and amplified before off-chip transfer and downstream nanopore sequencing. This workflow is sufficiently sensitive for obtaining different paired full-length antibody sequences from as little as five droplets, fulfilling the desired phenotype. Replacing physical sorting by specific sequencing overcomes a general bottleneck in droplet microfluidic screening and should be compatible with many more applications. [ABSTRACT FROM AUTHOR]
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- 2024
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18. Advanced Microfluidic Platform for Tumor Spheroid Formation and Cultivation Fabricated from OSTE+ Polymer.
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Panuška, Petr, Smejkal, Jiří, Štofik, Marcel, Žmudová, Zuzana, Španbauerová, Klára, Havlica, Jaromír, Harrandt, Václav, Vinopal, Stanislav, Aubrecht, Petr, and Malý, Jan
- Abstract
In the evolving landscape of cancer research, 3D cell cultures, particularly tumor cell spheroids, are increasingly preferred in drug screening due to their enhanced mimicry of in vivo tumor environments, especially in drug resistance aspects. However, the consistent formation of uniform spheroids and their precise manipulation remain complex challenges. Among various methodologies, droplet microfluidics emerges as a highly effective approach for tumor spheroid formation. This paper introduces a novel, multifaceted microfluidic system that streamlines the entire spheroid cultivation process: (i) generating tumor spheroids from cell suspensions within individual droplets, (ii) merging these droplets into a continuous aqueous phase once spheroid formation is complete, and (iii) transferring the spheroids to a specialized cultivation area within the chip, equipped with trapping elements for extended cultivation in perfusion mode. Remarkably, this process requires no hydrogel encapsulation or external handling, as all operations are conducted within the microfluidic chip. Fabricated from the innovative OSTE+ (off-stoichiometry thiol-ene epoxy) polymer, the chip is designed for repeated use. To show its efficacy, we successfully formed spheroids from MCF-7, GAMG, and U87 cell lines in our system and compared them with spheroids prepared by a traditional agarose microwell method. Additionally, our methodology has successfully enabled the in-chip release of spheroids from droplets, followed by their effective trapping for subsequent cultivation, a process we have exemplified with MCF-7 spheroids. To our knowledge, this research represents the first instance of a fully integrated droplet microfluidic platform achieving scaffoldless tumor spheroid formation and handling. Our method holds promise for improving high-throughput, automated procedures in the formation, transfer, and cultivation of tumor cell spheroids. [ABSTRACT FROM AUTHOR]
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- 2024
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19. Offsetting Dense Particle Sedimentation in Microfluidic Systems.
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Anyaduba, Tochukwu Dubem and Rodriguez-Manzano, Jesus
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MICROGELS ,FLUID dynamics ,FLOW meters ,IMAGE analysis ,PARTICLE tracks (Nuclear physics) - Abstract
Sedimentation is an undesirable phenomenon that complicates the design of microsystems that exploit dense microparticles as delivery tools, especially in biotechnological applications. It often informs the integration of continuous mixing modules, consequently impacting the system footprint, cost, and complexity. The impact of sedimentation is significantly worse in systems designed with the intent of particle metering or binary encapsulation in droplets. Circumventing this problem involves the unsatisfactory adoption of gel microparticles as an alternative. This paper presents two solutions—a hydrodynamic solution that changes the particle sedimentation trajectory relative to a flow-rate dependent resultant force, and induced hindered settling (i-HS), which exploits Richardson–Zaki (RZ) corrections of Stokes' law. The hydrodynamic solution was validated using a multi-well fluidic multiplexing and particle metering manifold. Computational image analysis of multiplex metering efficiency using this method showed an average reduction in well-to-well variation in particle concentration from 45% (Q = 1 mL/min, n = 32 total wells) to 17% (Q = 10 mL/min, n = 48 total wells). By exploiting a physical property (cloud point) of surfactants in the bead suspension in vials, the i-HS achieved a 58% reduction in the sedimentation rate. This effect results from the surfactant phase change, which increases the turbidity (transient increase in particle concentration), thereby exploiting the RZ theories. Both methods can be used independently or synergistically to eliminate bead settling in microsystems or to minimize particle sedimentation [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
20. Multimodal Imaging, Drug Delivery, and On‐Board Triggered Degradation in Soft Capsule Rolling Microrobots
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David Castellanos‐Robles, Raphaël C. L.‐M. Doineau, Azaam Aziz, Richard Nauber, Song Wu, Silvia Moreno, Konstantina Mitropoulou, Franziska Hebenstreit, and Mariana Medina‐Sánchez
- Subjects
droplet microfluidics ,in situ produced ultrasound/photoacoustic contrast agents ,local enzymatic degradation ,magnetic drug carriers ,medical soft microrobot ,polymersomes ,Computer engineering. Computer hardware ,TK7885-7895 ,Control engineering systems. Automatic machinery (General) ,TJ212-225 - Abstract
In the rapidly advancing field of medical microrobotics, designing robots capable of addressing various challenges—such as imaging, biodegradation, and multifunctionality—is crucial. Departing from conventional research that often focuses on isolated aspects of microrobot functionality, this study presents an innovative approach to comprehensive microrobot design. Soft capsule microrobots that integrate capabilities such as magnetic navigation, autonomous maneuverability, in situ biodegradation, biosafe imaging, and drug delivery are reported. These microrobots are fabricated within the range of 20–120 μm, with a notable throughput of ≈102–103 microrobots per second. Furthermore, their locomotion performance has been demonstrated to remain stable for a period exceeding 10 h, all while employing real‐time optical closed‐loop control. The incorporation of ultrasound contrast agents not only amplifies imaging resolution but also ensures imaging contrast stability in a biological environment for over a period of 3 h. Second, the intentional integration of enzyme‐loaded nanometric polymersomes establishes a self‐contained, biodegradable system, accentuating the microrobots’ capacity to degrade without the addition of high enzyme concentrations. This integrated approach lays the groundwork for minimally invasive treatments toward personalized and targeted medicine.
- Published
- 2024
- Full Text
- View/download PDF
21. Spherical Magnetic Fe-Alginate Microgels Fabricated by Droplet-Microfluidics Combining with an External Crosslinking Approach and the Study of Their pH Dependent Fe3+ Release Behaviors
- Author
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Chen, Jie, Yu, Run-Yu, Wang, Kai-Qi, Zhang, Zhe-Yu, Ardekani, Arezoo, and Hu, Yuan-Du
- Published
- 2024
- Full Text
- View/download PDF
22. Advances in the isolation, cultivation, and identification of gut microbes
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Meng-Qi Xu, Fei Pan, Li-Hua Peng, and Yun-Sheng Yang
- Subjects
Gut microbes ,Culturomics ,Microbial identification ,Droplet microfluidics ,Strain-level investigation ,Medicine (General) ,R5-920 ,Military Science - Abstract
Abstract The gut microbiome is closely associated with human health and the development of diseases. Isolating, characterizing, and identifying gut microbes are crucial for research on the gut microbiome and essential for advancing our understanding and utilization of it. Although culture-independent approaches have been developed, a pure culture is required for in-depth analysis of disease mechanisms and the development of biotherapy strategies. Currently, microbiome research faces the challenge of expanding the existing database of culturable gut microbiota and rapidly isolating target microorganisms. This review examines the advancements in gut microbe isolation and cultivation techniques, such as culturomics, droplet microfluidics, phenotypic and genomics selection, and membrane diffusion. Furthermore, we evaluate the progress made in technology for identifying gut microbes considering both non-targeted and targeted strategies. The focus of future research in gut microbial culturomics is expected to be on high-throughput, automation, and integration. Advancements in this field may facilitate strain-level investigation into the mechanisms underlying diseases related to gut microbiota.
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- 2024
- Full Text
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23. Thermostable in vitro transcription-translation compatible with microfluidic droplets
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Ana L. J. L. Ribeiro, Patricia Pérez-Arnaiz, Mercedes Sánchez-Costa, Lara Pérez, Marcos Almendros, Liisa van Vliet, Fabrice Gielen, Jesmine Lim, Simon Charnock, Florian Hollfelder, J. Eduardo González-Pastor, José Berenguer, and Aurelio Hidalgo
- Subjects
In vitro transcription and translation ,Thermus thermophiles ,Droplet microfluidics ,Cell-free protein expression ,Thermozymes ,Microbiology ,QR1-502 - Abstract
Abstract Background In vitro expression involves the utilization of the cellular transcription and translation machinery in an acellular context to produce one or more proteins of interest and has found widespread application in synthetic biology and in pharmaceutical biomanufacturing. Most in vitro expression systems available are active at moderate temperatures, but to screen large libraries of natural or artificial genetic diversity for highly thermostable enzymes or enzyme variants, it is instrumental to enable protein synthesis at high temperatures. Objectives Develop an in vitro expression system operating at high temperatures compatible with enzymatic assays and with technologies that enable ultrahigh-throughput protein expression in reduced volumes, such as microfluidic water-in-oil (w/o) droplets. Results We produced cell-free extracts from Thermus thermophilus for in vitro translation including thermostable enzymatic cascades for energy regeneration and a moderately thermostable RNA polymerase for transcription, which ultimately limited the temperature of protein synthesis. The yield was comparable or superior to other thermostable in vitro expression systems, while the preparation procedure is much simpler and can be suited to different Thermus thermophilus strains. Furthermore, these extracts have enabled in vitro expression in microfluidic droplets at high temperatures for the first time. Conclusions Cell-free extracts from Thermus thermophilus represent a simpler alternative to heavily optimized or pure component thermostable in vitro expression systems. Moreover, due to their compatibility with droplet microfluidics and enzyme assays at high temperatures, the reported system represents a convenient gateway for enzyme screening at higher temperatures with ultrahigh-throughput.
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- 2024
- Full Text
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24. Preparing HNS/n-Al heat-resistant microspheres with enhanced combustion performance using droplet microfluidic technology
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Bi-dong Wu, Yi Liu, Jia-hui Yang, Yun-yan Guo, Kai Han, Fan Wang, Zhong-ze Zhang, Chong-wei An, and Jing-yu Wang
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Droplet microfluidics ,Energetic microsphere ,Aluminized explosive ,Carbon cluster ,Combustion ,Chemical technology ,TP1-1185 - Abstract
Reducing the formation of large carbon clusters during the combustion of energetic materials (EMs) and improving their comprehensive performance hold great significance. With fluororubber (F2604) as a binder, this study prepared HNS/n-Al microspheres with different n-Al contents (5%, 10%, and 15%) using droplet microfluidic technology. Then, it characterized and tested the morphology, particle size distribution, dispersibility, crystal structure, thermal properties, mechanical sensitivity, and combustion behavior of the microspheres. The results show that the prepared microspheres had regular shapes, uniform particle sizes, and excellent dispersibility and contained more homogeneous components than physically mixed samples. Furthermore, the microspheres retained the crystal structures of the raw materials, enjoying high safety performance. The thermal analysis shows that HNS/n-Al microspheres had high heat resistance (thermal decomposition temperature: over 354 °C) and that a higher n-Al content was associated with more thorough thermal decomposition reactions of HNS (HNS: 83%, HNS/n-Al: 84%, 86%, and 93%). The ignition experiments show that the HNS/n-Al microspheres possessed excellent and stable combustion performance, as evidenced by more complete combustion reactions and significantly elevated energy release efficiency. Therefore, it is expected to achieve high-energy and high-speed responses of carbon-rich EMs and promote their practical applications.
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- 2024
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- View/download PDF
25. Amplification-Free COVID-19 Detection by Digital Droplet REVEALR
- Author
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Yang, Kefan and Chaput, John C
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Biochemistry and Cell Biology ,Bioinformatics and Computational Biology ,Biological Sciences ,Emerging Infectious Diseases ,Coronaviruses ,Infectious Diseases ,Genetics ,4.1 Discovery and preclinical testing of markers and technologies ,Infection ,Good Health and Well Being ,Humans ,COVID-19 ,SARS-CoV-2 ,DNA ,Catalytic ,Pandemics ,RNA ,Viral ,Nucleic Acid Amplification Techniques ,Sensitivity and Specificity ,RNA diagnostics ,droplet microfluidics ,REVEALR ,multicomponent DNAzyme ,Medicinal and Biomolecular Chemistry ,Biomedical Engineering ,Biochemistry and cell biology ,Bioinformatics and computational biology - Abstract
The COVID-19 pandemic, caused by the SARS-CoV-2 virus, exposed a pressing need for new public health tools for pathogen detection, disease diagnosis, and viral genotyping. REVEALR (RNA-encoded viral nucleic acid analyte reporter) is an isothermal DNAzyme-based point-of-care diagnostic that functions with a detection limit of ∼10 copies/μL when coupled with a preamplification step and can be utilized for viral genotyping of SARS-CoV-2 variants of concern through base pair mismatch recognition in a competitive binding format. Here, we describe an advanced REVEALR platform, termed digital droplet REVEALR (ddREVEALR), that can achieve direct viral detection and absolute sample quantitation utilizing a signal amplification strategy that relies on chemical modifications, DNAzyme multiplexing, and volume compression. Using an AI-assisted image-based readout, ddREVEALR was found to achieve 95% positive predictive agreement from a set of 20 nasal pharyngeal swabs collected at UCI Medical Center in Orange, California. We propose that the combination of amplification-free and protein-free analysis makes ddREVEALR a promising application for direct viral RNA detection of clinical samples.
- Published
- 2023
26. Droplet Microfluidics for High-Throughput Screening and Directed Evolution of Biomolecules.
- Author
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Vladisaljević, Goran T.
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NUCLEIC acids ,BIOMIMETIC materials ,HIGH throughput screening (Drug development) ,CATALYTIC RNA ,DELAY lines - Abstract
Directed evolution is a powerful technique for creating biomolecules such as proteins and nucleic acids with tailor-made properties for therapeutic and industrial applications by mimicking the natural evolution processes in the laboratory. Droplet microfluidics improved classical directed evolution by enabling time-consuming and laborious steps in this iterative process to be performed within monodispersed droplets in a highly controlled and automated manner. Droplet microfluidic chips can generate, manipulate, and sort individual droplets at kilohertz rates in a user-defined microchannel geometry, allowing new strategies for high-throughput screening and evolution of biomolecules. In this review, we discuss directed evolution studies in which droplet-based microfluidic systems were used to screen and improve the functional properties of biomolecules. We provide a systematic overview of basic on-chip fluidic operations, including reagent mixing by merging continuous fluid streams and droplet pairs, reagent addition by picoinjection, droplet generation, droplet incubation in delay lines, chambers and hydrodynamic traps, and droplet sorting techniques. Various microfluidic strategies for directed evolution using single and multiple emulsions and biomimetic materials (giant lipid vesicles, microgels, and microcapsules) are highlighted. Completely cell-free microfluidic-assisted in vitro compartmentalization methods that eliminate the need to clone DNA into cells after each round of mutagenesis are also presented. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
27. Targeted whole-genome recovery of single viral species in a complex environmental sample.
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Liyin Chen, Anqi Chen, Xinge Diana Zhang, Saenz Robles, Maria Teresa, Hee-Sun Han, Yi Xiao, Gao Xiao, Pipas, James M., and Weitz, David A.
- Subjects
- *
VIRAL genomes , *VIRAL ecology , *SV40 (Virus) , *WHOLE genome sequencing , *ENVIRONMENTAL sampling - Abstract
Characterizing unknown viruses is essential for understanding viral ecology and preparing against viral outbreaks. Recovering complete genome sequences from environmental samples remains computationally challenging using metagenomics, especially for low-abundance species with uneven coverage. We present an experimental method for reliably recovering complete viral genomes from complex environmental samples. Individual genomes are encapsulated into droplets and amplified using multiple displacement amplification. A unique gene detection assay, which employs an RNA-based probe and an exonuclease, selectively identifies droplets containing the target viral genome. Labeled droplets are sorted using a microfluidic sorter, and genomes are extracted for sequencing. We demonstrate this method's efficacy by spiking two known viral genomes, Simian virus 40 (SV40, 5,243 bp) and Human Adenovirus 5 (HAd5, 35,938 bp), into a sewage sample with a final abundance in the droplets of around 0.1% and 0.015%, respectively. We achieve 100% recovery of the complete sequence of the spiked-in SV40 genome with uniform coverage distribution. For the larger HAd5 genome, we cover approximately 99.4% of its sequence. Notably, genome recovery is achieved with as few as one sorted droplet, which enables the recovery of any desired genomes in complex environmental samples, regardless of their abundance. This method enables single-genome whole-genome amplification and targeting characterizations of rare viral species and will facilitate our ability to access the mutational profile in single-virus genomes and contribute to an improved understanding of viral ecology. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
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28. XPORT ENTRAP: A droplet microfluidic platform for enhanced DNA transfer between microbial species.
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Wippold, Jose A., Chu, Monica, Renberg, Rebecca, Li, Yuwen, Adams, Bryn, and Han, Arum
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MICROFLUIDIC devices , *HORIZONTAL gene transfer , *LABS on a chip , *MOLECULAR biology , *DNA , *GENETIC transformation , *SYNTHETIC biology - Abstract
A significant hurdle for the widespread implementation and use of synthetic biology is the challenge of highly efficient introduction of DNA into microorganisms. This is especially a barrier for the utilization of non-model organisms and/or novel chassis species for a variety of applications, ranging from molecular biology to biotechnology and biomanufacturing applications. Common approaches to episomal and chromosomal gene editing, which employ techniques such as chemical competence and electroporation, are typically only amenable to a small subset of microbial species while leaving the vast majority of microorganisms in nature genetically inaccessible. To address this challenge, we have employed the previously described B. subtilis broad-host conjugation strain, XPORT, which was modularly designed for loading DNA cargo and conjugating such DNA into recalcitrant microbes. In this current work, we have leveraged and adapted the XPORT strain for use in a droplet microfluidic platform to enable increased efficiency of conjugation-based DNA transfer. The system named DNA ENTRAP (DNA ENhanced TRAnsfer Platform) utilizes cell-encapsulated water-in-oil emulsion droplets as pico-liter-volume bioreactors that allows controlled contacts between the donor and receiver cells within the emulsion bioreactor. This allowed enhanced XPORT-mediated genetic transfer over the current benchtop XPORT process, demonstrated using two different Bacillus subtilis strains (donor and receiver), as well as increased throughput (e.g., number of successfully conjugated cells) due to the automated assay steps inherent to microfluidic lab-on-a-chip systems. DNA ENTRAP paves the way for a streamlined automation of culturing and XPORT-mediated genetic transfer processes as well as future high-throughput cell engineering and screening applications. • Developed a droplet microfluidic device (DNA ENTRAP) for microbial engineering. • Demonstrated horizontal gene transfer efficiency increases over 200%. • Encapsulation of donor and recipient in a microdroplet enhances gene transfer. • Microfluidic-adapted SynBio tool for the domestication of untapped microbes. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
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29. Fabrication of Crescent Shaped Microparticles for Particle Templated Droplet Formation.
- Author
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Yang, Yimin, Vagin, Sergei I., Rieger, Bernhard, and Destgeer, Ghulam
- Subjects
- *
NANOGELS , *PHASE separation , *POLYMER fractionation , *FLUORESCENT dyes , *PHASE diagrams - Abstract
Crescent‐shaped hydrogel microparticles are shown to template uniform volume aqueous droplets upon simple mixing with aqueous and oil media for various bioassays. This emerging "lab on a particle" technique requires hydrogel particles with tunable material properties and dimensions. The crescent shape of the particles is attained by aqueous two‐phase separation of polymers followed by photopolymerization of the curable precursor. In this work, the phase separation of poly(ethylene glycol) diacrylate (PEGDA, Mw 700) and dextran (Mw 40 000) for tunable manufacturing of crescent‐shaped particles is investigated. The particles' morphology is precisely tuned by following a phase diagram, varying the UV intensity, and adjusting the flow rates of various streams. The fabricated particles with variable dimensions encapsulate uniform aqueous droplets upon mixing with an oil phase. The particles are fluorescently labeled with red and blue emitting dyes at variable concentrations to produce six color‐coded particles. The blue fluorescent dye shows a moderate response to the pH change. The fluorescently labeled particles are able to tolerate an extremely acidic solution (pH 1) but disintegrate within an extremely basic solution (pH 14). The particle‐templated droplets are able to effectively retain the disintegrating particle and the fluorescent signal at pH 14. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
30. Droplet digital molecular beacon-LAMP assay via pico-injection for ultrasensitive detection of pathogens.
- Author
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Ma, Zhiyuan, Ma, Mengshao, Cao, Xiaobao, Jiang, Yuyang, and Gao, Dan
- Subjects
- *
MICROFLUIDICS , *ESCHERICHIA coli O157:H7 , *MICROFLUIDIC devices , *PATHOGENIC microorganisms , *DETECTION limit , *ESCHERICHIA coli - Abstract
A pico-injection-aided digital droplet detection platform is presented that integrates loop-mediated isothermal amplification (LAMP) with molecular beacons (MBs) for the ultrasensitive and quantitative identification of pathogens, leveraging the sequence-specific detection capabilities of MBs. The microfluidic device contained three distinct functional units including droplet generation, pico-injection, and droplet counting. Utilizing a pico-injector, MBs are introduced into each droplet to specifically identify LAMP amplification products, thereby overcoming issues related to temperature incompatibility. Our methodology has been validated through the quantitative detection of Escherichia coli, achieving a detection limit as low as 9 copies/μL in a model plasmid containing the malB gene and 3 CFU/μL in a spiked milk sample. The total analysis time was less than 1.5 h. The sensitivity and robustness of this platform further demonstrated the potential for rapid pathogen detection and diagnosis, particularly when integrated with cutting-edge microfluidic technologies. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
31. مطالعه عددی تشکیل قطرات مرکب در یک سیستم میکرو سیال مویین شیشه ای سه فازی.
- Author
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سعید پوراحمدیان and محمد علی بیجارچی
- Subjects
CONTACT angle ,REGIME change ,CELL culture ,COSMETICS industry ,NOZZLES - Abstract
Copyright of Modares Mechanical Engineering is the property of Tarbiat Modares University Press and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
- Published
- 2024
32. CellMag‐CARWash: A High Throughput Droplet Microfluidic Device for Live Cell Isolation and Single Cell Applications.
- Author
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Rupp, Brittany T., Cook, Claire D., Purcell, Emma A., Pop, Matei, Radomski, Abigail E., Mesyngier, Nicolas, Bailey, Ryan C., and Nagrath, Sunitha
- Subjects
CELL separation ,MICROFLUIDIC devices ,CELL populations ,EXTRACELLULAR vesicles ,POSITIVE systems - Abstract
The recent push toward understanding an individual cell's behavior and identifying cellular heterogeneity has created an unmet need for technologies that can probe live cells at the single‐cell level. Cells within a population are known to exhibit heterogeneous responses to environmental cues. These differences can lead to varied cellular states, behavior, and responses to therapeutics. Techniques are needed that are not only capable of processing and analyzing cellular populations at the single cell level, but also have the ability to isolate specific cell populations from a complex sample at high throughputs. The new CellMag‐Coalesce‐Attract‐Resegment Wash (CellMag‐CARWash) system combines positive magnetic selection with droplet microfluidic devices to isolate cells of interest from a mixture with >93% purity and incorporate treatments within individual droplets to observe single cell biological responses. This workflow is shown to be capable of probing the single cell extracellular vesicle (EV) secretion of MCF7 GFP cells. This article reports the first measurement of β‐Estradiol's effect on EV secretion from MCF7 cells at the single cell level. Single cell processing revealed that MCF7 GFP cells possess a heterogeneous response to β‐Estradiol stimulation with a 1.8‐fold increase relative to the control. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
33. Advancing Point-of-Care Applications with Droplet Microfluidics: From Single-Cell to Multicellular Analysis.
- Author
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Sharkey, Christina, White, Rachel, Finocchiaro, Michael, Thomas, Judene, Estevam, Jose, and Konry, Tania
- Abstract
Recent advances in single-cell and multicellular microfluidics technology have provided powerful tools for studying cancer biology and immunology. The ability to create controlled microenvironments, perform high-throughput screenings, and monitor cellular interactions at the single-cell level has significantly advanced our understanding of tumor biology and immune responses. We discuss cutting-edge multicellular and single-cell microfluidic technologies and methodologies utilized to investigate cancer–immune cell interactions and assess the effectiveness of immunotherapies. We explore the advantages and limitations of the wide range of 3D spheroid and single-cell microfluidic models recently developed, highlighting the various approaches in device generation and applications in immunotherapy screening for potential opportunities for point-of-care approaches. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
34. Advances in the isolation, cultivation, and identification of gut microbes.
- Author
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Xu, Meng-Qi, Pan, Fei, Peng, Li-Hua, and Yang, Yun-Sheng
- Abstract
The gut microbiome is closely associated with human health and the development of diseases. Isolating, characterizing, and identifying gut microbes are crucial for research on the gut microbiome and essential for advancing our understanding and utilization of it. Although culture-independent approaches have been developed, a pure culture is required for in-depth analysis of disease mechanisms and the development of biotherapy strategies. Currently, microbiome research faces the challenge of expanding the existing database of culturable gut microbiota and rapidly isolating target microorganisms. This review examines the advancements in gut microbe isolation and cultivation techniques, such as culturomics, droplet microfluidics, phenotypic and genomics selection, and membrane diffusion. Furthermore, we evaluate the progress made in technology for identifying gut microbes considering both non-targeted and targeted strategies. The focus of future research in gut microbial culturomics is expected to be on high-throughput, automation, and integration. Advancements in this field may facilitate strain-level investigation into the mechanisms underlying diseases related to gut microbiota. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
35. Uniform sized cancer spheroids production using hydrogel-based droplet microfluidics: a review.
- Author
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Sungjin Kim, Lam, Po Yi, Jayaraman, Arul, and Han, Arum
- Abstract
Three-dimensional (3D) cell culture models have been extensively utilized in various mechanistic studies as well as for drug development studies as superior in vitro platforms than conventional two-dimensional (2D) cell culture models. This is especially the case in cancer biology, where 3D cancer models, such as spheroids or organoids, have been utilized extensively to understand the mechanisms of cancer development. Recently, many sophisticated 3D models such as organ-on-a-chip models are emerging as advanced in vitro models that can more accurately mimic the in vivo tissue functions. Despite such advancements, spheroids are still considered as a powerful 3D cancer model due to the relatively simple structure and compatibility with existing laboratory instruments, and also can provide orders of magnitude higher throughput than complex in vitro models, an extremely important aspects for drug development. However, creating well-defined spheroids remain challenging, both in terms of throughputs in generation as well as reproducibility in size and shape that can make it challenging for drug testing applications. In the past decades, droplet microfluidics utilizing hydrogels have been highlighted due to their potentials. Importantly, core-shell structured gel droplets can avoid spheroid-to-spheroid adhesion that can cause large variations in assays while also enabling long-term cultivation of spheroids with higher uniformity by protecting the core organoid area from external environment while the outer porous gel layer still allows nutrient exchange. Hence, core-shell gel droplet-based spheroid formation can improve the predictivity and reproducibility of drug screening assays. This review paper will focus on droplet microfluidics-based technologies for cancer spheroid production using various gel materials and structures. In addition, we will discuss emerging technologies that have the potential to advance the production of spheroids, prospects of such technologies, and remaining challenges. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
36. Microfluidic Technologies and Platforms for Protein Crystallography
- Author
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Maeki, Masatoshi, Tokeshi, Manabu, Vo-Dinh, Tuan, Series Editor, and Tokeshi, Manabu, editor
- Published
- 2024
- Full Text
- View/download PDF
37. Droplet Microfluidic Systems for Multistep Single-Cell Sequencing Assays
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Olszewska, Zofia, Pyzik, Adam, Malkowski, Marcin, Kaminski, Tomasz S., Vo-Dinh, Tuan, Series Editor, and Tokeshi, Manabu, editor
- Published
- 2024
- Full Text
- View/download PDF
38. Application of Microfluidics in Drug Development
- Author
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Parhizkar, Maryam, Wang, Fanjin, Anna, Tsitouridou, Tsaoulidis, Dimitrios, Salomon, Claudio, Series Editor, Zavod, Robin, Founding Editor, Lamprou, Dimitrios A., editor, and Weaver, Edward, editor
- Published
- 2024
- Full Text
- View/download PDF
39. Automated and miniaturized screening of antibiotic combinations via robotic-printed combinatorial droplet platform
- Author
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Fangchi Shao, Hui Li, Kuangwen Hsieh, Pengfei Zhang, Sixuan Li, and Tza-Huei Wang
- Subjects
Droplet microfluidics ,Antimicrobial resistance ,Antibiotic combinatorial screening ,Contact printing ,Robotics ,Pharmaceutical analysis ,Therapeutics. Pharmacology ,RM1-950 - Abstract
Antimicrobial resistance (AMR) has become a global health crisis in need of novel solutions. To this end, antibiotic combination therapies, which combine multiple antibiotics for treatment, have attracted significant attention as a potential approach for combating AMR. To facilitate advances in antibiotic combination therapies, most notably in investigating antibiotic interactions and identifying synergistic antibiotic combinations however, there remains a need for automated high-throughput platforms that can create and examine antibiotic combinations on-demand, at scale, and with minimal reagent consumption. To address these challenges, we have developed a Robotic-Printed Combinatorial Droplet (RoboDrop) platform by integrating a programmable droplet microfluidic device that generates antibiotic combinations in nanoliter droplets in automation, a robotic arm that arranges the droplets in an array, and a camera that images the array of thousands of droplets in parallel. We further implement a resazurin-based bacterial viability assay to accelerate our antibiotic combination testing. As a demonstration, we use RoboDrop to corroborate two pairs of antibiotics with known interactions and subsequently identify a new synergistic combination of cefsulodin, penicillin, and oxacillin against a model E. coli strain. We therefore envision RoboDrop becoming a useful tool to efficiently identify new synergistic antibiotic combinations toward combating AMR.
- Published
- 2024
- Full Text
- View/download PDF
40. Droplet microfluidics for time-resolved serial crystallography
- Author
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Jack Stubbs, Theo Hornsey, Niall Hanrahan, Luis Blay Esteban, Rachel Bolton, Martin Malý, Shibom Basu, Julien Orlans, Daniele de Sanctis, Jung-uk Shim, Patrick D. Shaw Stewart, Allen M. Orville, Ivo Tews, and Jonathan West
- Subjects
droplet microfluidics ,crystal miniaturization ,micromixing ,time-resolved serial crystallography ,Crystallography ,QD901-999 - Abstract
Serial crystallography requires large numbers of microcrystals and robust strategies to rapidly apply substrates to initiate reactions in time-resolved studies. Here, we report the use of droplet miniaturization for the controlled production of uniform crystals, providing an avenue for controlled substrate addition and synchronous reaction initiation. The approach was evaluated using two enzymatic systems, yielding 3 µm crystals of lysozyme and 2 µm crystals of Pdx1, an Arabidopsis enzyme involved in vitamin B6 biosynthesis. A seeding strategy was used to overcome the improbability of Pdx1 nucleation occurring with diminishing droplet volumes. Convection within droplets was exploited for rapid crystal mixing with ligands. Mixing times of
- Published
- 2024
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- View/download PDF
41. High-throughput screening of gold nanoparticle synthesis parameters in droplet microfluidics.
- Author
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Egil, Nikita V., Zagrebaev, Alexander D., Medvedev, Pavel V., Alexandrov, Alexander A., Bagliy, Anton P., Chapek, Sergey V., Guda, Alexander A., and Soldatov, Alexander V.
- Subjects
- *
GOLD nanoparticles , *NANOPARTICLE synthesis , *HIGH throughput screening (Drug development) , *MEDICAL screening , *THREE-dimensional printing - Abstract
[Display omitted] A 3D printed microreactor and UV-VIS spectral diagnostic system were adapted for an automatic protocol for screening parameters of citrate-based droplet synthesis of gold nanoparticles. Using this system, 27 combinations of synthesis parameters were screened within an hour and data were collected from more than 1000 individual droplets with nanoparticles. The developed system allows both filtering the spectra of target droplets and conducting a comprehensive multivariate analysis of all key descriptors. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
42. Highly sensitive absorbance measurement using droplet microfluidics integrated with an oil extraction and long pathlength detection flow cell.
- Author
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Lu, Bingyuan, Lunn, James, Nightingale, Adrian M., Niu, Xize, Moreno, Sergio Quintero, and Toprakcioglu, Zenon
- Subjects
- *
AQUACULTURE , *ANTIBIOTICS , *OXYTETRACYCLINE , *PROBIOTICS , *IMMUNOREGULATION - Abstract
In droplet microfluidics, UV-Vis absorption spectroscopy along with colorimetric assays have been widely used for chemical and biochemical analysis. However, the sensitivity of the measurement can be limited by the short optical pathlength. Here we report a novel design to enhance the sensitivity by removing oil and converting the droplets into a single-phase aqueous flow, which can be measured within a U-shape channel with long optical pathlength. The flow cells were fabricated via 3D printing. The calibration results have demonstrated complete oil removal and effective optical pathlengths similar to the designed channel lengths (from 5 to 20 mm). The flow cell was further employed in a droplet microfluidic-based phosphate sensing system. The measured phosphate levels displayed excellent consistency with data obtained from traditional UV spectroscopy analysis. This flow cell design overcomes the limitations of short optical pathlengths in droplet microfluidics and has the potential to be used for in situ and continuous monitoring. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
43. Merged and alternating droplets generation in double T-junction microchannels using symmetrically inserted capillaries.
- Author
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Shen, Feng, Zhang, Yuedong, Li, Chunyou, Pang, Yan, and Liu, Zhaomiao
- Abstract
In this work, merged and alternating droplets generated in a microfluidic double T-junction are investigated using experiments and numerical simulations. The double T-junction is constructed by symmetrically inserting two capillaries into a microfluidic chip at specific positions. We explore the effects of the two-phase flow rate fraction, capillary tip distance (30 μm, 60 μm, and 200 μm), and fluid properties on droplet formation phenomena. Detailed observations reveal four distinct regimes during the dynamic evolution of the two-phase interface morphology: merged state, stable alternating droplets, droplet pairs, and jetting. Two phase diagrams are obtained to demonstrate that interfacial tension and dispersed phase viscosity significantly influence these regimes. Moreover, we find that as the flow rate fraction increases from 0.054 to 0.286, the length of generated droplets increases from 156 to 789 μm; we provide a theoretical prediction formula for dimensionless droplet length accordingly. Additionally, our simulations show fluctuating pressure in dispersed flows throughout the process of droplet generation. The simulated pressure in the dispersed flows fluctuates during the droplet generation process. The understanding of the underlying physics of the capillary-based double T-junction contributes valuable insights for various related applications. Highlights: An easy capillary-based double T-junction for droplet generation. Merged and alternating droplets are generated. Four regimes are observed as the merged, stable alternating droplet, droplet pairs, and jetting. Effects of two-phase flow rates, capillary tip distances, and fluid properties are investigated. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
44. Exploring the stability of single emulsion created by microfluidics and its use in the production of core–shell microparticles.
- Author
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Oveysi, Mehrnaz, Bazargan, Vahid, Nejat, Amir, and Marengo, Marco
- Abstract
This study introduces an innovative method aimed at achieving exceptional stability in emulsions. The primary focus is on re-emulsifying precisely controlled and uniform initial single emulsions, generated by microfluidic devices, to produce single-core double emulsions and core–shell microparticles. Departing from traditional approaches, our method employs a unique combination of advanced Two-level fractional factorial design and numerical simulation. These tools are utilized to discern and optimize critical parameters necessary for the formation of highly monodispersed stable single emulsions and their subsequent transformation into double emulsions. Correlations are established to estimate the size and stability of the primary single emulsion based on immiscible phase flow rate ratio and surfactant concentration. These correlations provide a comprehensive understanding that facilitates the intentional development of desired water-in-oil emulsions. The proposed microfluidic paradigm shows promise for the controlled and efficient production of single-core double emulsions, with broad applications in Pharmaceuticals, Food, and Cosmetics. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
45. A Label-Free Droplet Sorting Platform Integrating Dielectrophoretic Separation for Estimating Bacterial Antimicrobial Resistance.
- Author
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Yan, Jia-De, Yang, Chiou-Ying, Han, Arum, and Wu, Ching-Chou
- Subjects
DRUG resistance in bacteria ,DRUG resistance in microorganisms ,ESCHERICHIA coli ,CELL separation ,DIELECTROPHORESIS ,DRUG efficacy ,ELECTRIC fields - Abstract
Antimicrobial resistance (AMR) has become a crucial global health issue. Antibiotic-resistant bacteria can survive after antibiotic treatments, lowering drug efficacy and increasing lethal risks. A microfluidic water-in-oil emulsion droplet system can entrap microorganisms and antibiotics within the tiny bioreactor, separate from the surroundings, enabling independent assays that can be performed in a high-throughput manner. This study presents the development of a label-free dielectrophoresis (DEP)-based microfluidic platform to sort droplets that co-encapsulate Escherichia coli (E. coli) and ampicillin (Amp) and droplets that co-encapsulate Amp-resistant (AmpR) E. coli with Amp only based on the conductivity-dependent DEP force (F
DEP ) without the assistance of optical analyses. The 9.4% low conductivity (LC) Luria–Bertani (LB) broth diluted with 170 mM mannitol can maintain E. coli and AmpR E. coli growth for 3 h and allow Amp to kill almost all E. coli, which can significantly increase the LCLB conductivity by about 100 μS/cm. Therefore, the AmpR E. coli/9.4%LCLB/Amp where no cells are killed and the E. coli/9.4%LCLB/Amp-containing droplets where most of the cells are killed can be sorted based on this conductivity difference at an applied electric field of 2 MHz and 100 Vpp that generates positive FDEP . Moreover, the sorting ratio significantly decreased to about 50% when the population of AmpR E. coli was equal to or higher than 50% in droplets. The conductivity-dependent DEP-based sorting platform exhibits promising potential to probe the ratio of AmpR E. coli in an unknown bacterial sample by using the sorting ratio as an index. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
46. High‐Throughput Single‐Cell, Single‐Mitochondrial DNA Assay Using Hydrogel Droplet Microfluidics.
- Author
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Park, Juhwan, Kadam, Parnika S., Atiyas, Yasemin, Chhay, Bonirath, Tsourkas, Andrew, Eberwine, James H., and Issadore, David A.
- Subjects
- *
MITOCHONDRIAL DNA , *MICROFLUIDICS , *HYDROGELS , *CELL populations , *DNA , *PROTEIN analysis - Abstract
There is growing interest in understanding the biological implications of single cell heterogeneity and heteroplasmy of mitochondrial DNA (mtDNA), but current methodologies for single‐cell mtDNA analysis limit the scale of analysis to small cell populations. Although droplet microfluidics have increased the throughput of single‐cell genomic, RNA, and protein analysis, their application to sub‐cellular organelle analysis has remained a largely unsolved challenge. Here, we introduce an agarose‐based droplet microfluidic approach for single‐cell, single‐mtDNA analysis, which allows simultaneous processing of hundreds of individual mtDNA molecules within >10,000 individual cells. Our microfluidic chip encapsulates individual cells in agarose beads, designed to have a sufficiently dense hydrogel network to retain mtDNA after lysis and provide a robust scaffold for subsequent multi‐step processing and analysis. To mitigate the impact of the high viscosity of agarose required for mtDNA retention on the throughput of microfluidics, we developed a parallelized device, successfully achieving ~95 % mtDNA retention from single cells within our microbeads at >700,000 drops/minute. To demonstrate utility, we analyzed specific regions of the single‐mtDNA using a multiplexed rolling circle amplification (RCA) assay. We demonstrated compatibility with both microscopy, for digital counting of individual RCA products, and flow cytometry for higher throughput analysis. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
47. High‐Throughput Host–Microbe Single‐Cell RNA Sequencing Reveals Ferroptosis‐Associated Heterogeneity during Acinetobacter baumannii Infection.
- Author
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Meng, Hongen, Zhang, Tianyu, Wang, Zhang, Zhu, Yuyi, Yu, Yingying, Chen, Hangfei, Chen, Jiaye, Wang, Fudi, Yu, Yunsong, Hua, Xiaoting, and Wang, Yongcheng
- Subjects
- *
ACINETOBACTER infections , *ACINETOBACTER baumannii , *RNA sequencing , *BACTERIAL RNA , *BACTERIAL cells , *HETEROGENEITY - Abstract
Interactions between host and bacterial cells are integral to human physiology. The complexity of host–microbe interactions extends to different cell types, spatial aspects, and phenotypic heterogeneity, requiring high‐resolution approaches to capture their full complexity. The latest breakthroughs in single‐cell RNA sequencing (scRNA‐seq) have opened up a new era of studies in host–pathogen interactions. Here, we first report a high‐throughput cross‐species dual scRNA‐seq technology by using random primers to simultaneously capture both eukaryotic and bacterial RNAs (scRandom‐seq). Using reference cells, scRandom‐seq can detect individual eukaryotic and bacterial cells with high throughput and high specificity. Acinetobacter baumannii (A.b) is a highly opportunistic and nosocomial pathogen that displays resistance to many antibiotics, posing a significant threat to human health, calling for discoveries and treatment. In the A.b infection model, scRandom‐seq witnessed polarization of THP‐1 derived‐macrophages and the intracellular A.b‐induced ferroptosis‐stress in host cells. The inhibition of ferroptosis by Ferrostatin‐1 (Fer‐1) resulted in the improvement of cell vitality and resistance to A.b infection, indicating the potential to resist related infections. scRandom‐seq provides a high‐throughput cross‐species dual single‐cell RNA profiling tool that will facilitate future discoveries in unraveling the complex interactions of host–microbe interactions in infection systems and tumor micro‐environments. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
48. Computational Design of Synthetic Optical Barcodes in Microdroplets.
- Author
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Kawasaki, Fumiko, Mimori, Takahiro, Mori, Yuka, Aburatani, Hiroyuki, Yachie, Nozomu, Sato, Issei, and Ota, Sadao
- Subjects
- *
MICRODROPLETS , *NANOGELS , *GENETIC barcoding , *BAR codes , *CELL analysis - Abstract
Barcodes are useful for identifying objects across time, space, and information modalities. However, materializing and decoding optical and multimodal barcodes on microscopic objects remains difficult despite the increasing need for multiplexed cell analysis. Here, a computational design of randomly combinatorial is presented, yet decodable barcodes in microdroplets. The design is based on a novel Real2Sim2Real framework: it first collects experimental images of optically distinct microparticles, then simulates massive combinatorial images by randomly assembling the imaged particles to train a neural network‐based decoder. It is demonstrated that the decoder, even though trained via simulation, accurately identifies the randomly assembled particles in real hydrogel microdroplets. It also shows that the microdroplets with an additional DNA barcoding functionality are applicable to individually link independently measured microscopic images and transcriptome profiles of pooled single cells. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
49. Peptide Nucleic Acid Clamp‐Assisted Photothermal Multiplexed Digital PCR for Identifying SARS‐CoV‐2 Variants of Concern.
- Author
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Zhang, Lexiang, Parvin, Rokshana, Lin, Siyue, Chen, Mingshuo, Zheng, Ruixuan, Fan, Qihui, and Ye, Fangfu
- Subjects
- *
PEPTIDE nucleic acids , *SARS-CoV-2 , *SARS-CoV-2 Delta variant , *SARS-CoV-2 Omicron variant , *SINGLE nucleotide polymorphisms - Abstract
The unprecedented demand for variants diagnosis in response to the COVID‐19 epidemic has brought the spotlight onto rapid and accurate detection assays for single nucleotide polymorphisms (SNPs) at multiple locations. However, it is still challenging to ensure simplicity, affordability, and compatibility with multiplexing. Here, a novel technique is presented that combines peptide nucleic acid (PNA) clamps and near‐infrared (NIR)‐driven digital polymerase chain reaction (dPCR) to identify the Omicron and Delta variants. This is achieved by simultaneously identifying highly conserved mutated signatures at codons 19, 614, and 655 of the spike protein gene. By microfluidically introducing graphene‐oxide‐nanocomposite into the assembled gelatin microcarriers, they achieved a rapid temperature ramping‐up rate and switchable gel‐to‐sol phase transformation synchronized with PCR activation under NIR irradiation. Two sets of duplex PCR reactions, each classifying respective PNA probes, are emulsified in parallel and illuminated together using a homemade vacuum‐based droplet generation device and a programmable NIR control module. This allowed for selective amplification of mutant sequences due to single‐base‐pair mismatch with PNA blockers. Sequence‐recognized bioreactions and fluorescent‐color scoring enabled quick identification of variants. This technique achieved a detection limit of 5,100 copies and a 5‐fold quantitative resolution, which is promising to unfold minor differences and dynamic changes. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
50. Droplet-based cell-laden microgels for high-throughput analysis.
- Author
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Li, Xiang, Zhao, Danshan, Wang, Yuetong, and Huang, He
- Abstract
Cell-laden droplet microfluidics has revolutionized bulk biochemical analysis by offering compartmentalized microreactors for individual cells, but downstream operations of regular aqueous droplets are limited. Hydrogel matrix can provide a rigid scaffold for long-term culture of eukaryotic and prokaryotic cells, and can support several manipulations, facilitating subsequent high-throughput analysis of cellular heterogeneity. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
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