Lab on a Chip最新文献

{"title":"Parallel DLD Microfluidics for Chloroplast Isolation and Sorting","authors":"Oriana Gerallin Chavez-Pineda, Pablo E. Guevara-Pantoja, Victor Marin, Gabriel Arturo Caballero-Robledo, Luis David Patino-Lopez, Daniel A May-Arrioja, Clelia De-la-Peña, Jose L. Garcia-Cordero","doi":"10.1039/d5lc00348b","DOIUrl":"https://doi.org/10.1039/d5lc00348b","url":null,"abstract":"Chloroplasts are characteristic organelles of plant cells, essential for photosynthesis and various other metabolic processes, including amino acid, lipid, and hormone biosynthesis. Beyond their classical functions, chloroplasts have emerged as promising targets in biotechnology, particularly in therapeutic applications and biofuel production. However, their isolation remains technically challenging due to the limitations of conventional methods, which typically require complex protocols, specialized equipment, and trained personnel. Here, we present a microfluidic-based platform that enables size-based chloroplast separation using deterministic lateral displacement (DLD). Our device integrates four parallel DLD arrays, each with a distinct critical diameter (CD). This configuration enables bandpass filtering and allows the simultaneous isolation of chloroplasts of various sizes within a single device. Shared inlets and uniform flow conditions across all arrays enhance reproducibility compared to conventional techniques. Unlike traditional sucrose density gradients, which lack precise size-based separation, our system achieves separation efficiencies of 50-85% for chloroplasts ranging from 3 to 8 µm, with recovered fractions having purities of 17-66%. This platform provides a rapid, automated, and scalable solution for chloroplast isolation, with significant potential applications in plant research, biotechnology, and synthetic biology.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"45 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144237778","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An <i>in vivo</i> mimetic liver-lobule-chip (LLoC) for stem cell maturation, and zonation of hepatocyte-like cells on chip.","authors":"Philip Dalsbecker, Siiri Suominen, Muhammad Asim Faridi, Reza Mahdavi, Julia Johansson, Charlotte Hamngren Blomqvist, Mattias Goksör, Katriina Aalto-Setälä, Leena E Viiri, Caroline B Adiels","doi":"10.1039/d4lc00509k","DOIUrl":"https://doi.org/10.1039/d4lc00509k","url":null,"abstract":"<p><p><i>In vitro</i> cell culture models play a crucial role in preclinical drug discovery. To achieve optimal culturing environments and establish physiologically relevant organ-specific conditions, it is imperative to replicate <i>in vivo</i> scenarios when working with primary or induced pluripotent cell types. However, current approaches to recreating <i>in vivo</i> conditions and generating relevant 3D cell cultures still fall short. In this study, we validate a liver-lobule-chip (LLoC) containing 21 artificial liver lobules, each representing the smallest functional unit of the human liver. The LLoC facilitates diffusion-based perfusion <i>via</i> sinusoid-mimetic structures, providing physiologically relevant shear stress exposure and radial nutrient concentration gradients within each lobule. We demonstrate the feasibility of long term cultures (up to 14 days) of viable and functional HepG2 cells in a 3D discoid tissue structure, serving as initial proof of concept. Thereafter, we successfully differentiate sensitive, human induced pluripotent stem cell (iPSC)-derived cells into hepatocyte-like cells over a period of 20 days on-chip, exhibiting advancements in maturity compared to traditional 2D cultures. Further, hepatocyte-like cells cultured in the LLoC exhibit zonated protein expression profiles, indicating the presence of metabolic gradients characteristic of liver lobules. Our results highlight the suitability of the LLoC for long-term discoid tissue cultures, specifically for iPSCs, and their differentiation in a perfused environment. We envision the LLoC as a starting point for more advanced <i>in vitro</i> models, allowing for the combination of multiple liver cell types to create a comprehensive liver model for disease-onchip studies. Ultimately, when combined with stem cell technology, the LLoC offers a promising and robust on-chip liver model that serves as a viable alternative to primary hepatocyte cultures-ideally suited for preclinical drug screening and personalized medicine applications.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144245375","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Microfluidic Device for Passive Separation of Platelet-Rich Plasma from Whole Blood","authors":"Pablo E. Guevara-Pantoja, Yara Alvarez-Brana, Jon Mercader Ruiz, Fernando Benito-Lopez, Lourdes Basabe-Desmonts","doi":"10.1039/d5lc00362h","DOIUrl":"https://doi.org/10.1039/d5lc00362h","url":null,"abstract":"We present a microfluidic device for separating Platelet-Rich Plasma (PRP) from whole blood, addressing key limitations in current technologies. Unlike existing methods that require complex fabrication and expensive materials, our approach uses a CO2 laser cutter to fabricate acrylic layers bonded with pressure-sensitive adhesives, making it cost-effective and simple. Operating via gravity sedimentation, the device captures blood cells in multiple trenches and processes 1 mL of whole blood—significantly more than previous sedimentation devices. To prevent bubble formation, we incorporated a hydrophilic surface at the trench bottoms, ensuring reliable PRP separation. We analyzed three trench geometries to optimize plasma yield and quality. Our device processes blood in 45 minutes, yielding ~300 µL of plasma with at least a 2-fold platelet concentration increase. Red and white blood cell removal purities are 98% and 96%, respectively. Flow simulations optimized shear rates to improve sedimentation. Only 8.2% of the total platelets were activated, compared to 31% in the centrifugation method. This combination of simplicity, cost-efficiency, and effective platelet preservation enables a low-cost method for obtaining high-quality PRP for clinical research and therapy.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"18 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144228823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Size-selective sorting of kaolinite micro/nanoflakes via microfluidic filtration for wound hemostasis†","authors":"Guangyao Li, Liang Wan, Ying Chen, Xuming Zhang, Aidong Tang and Huaming Yang","doi":"10.1039/D5LC00274E","DOIUrl":"10.1039/D5LC00274E","url":null,"abstract":"<p >Kaolinite, a natural micro/nano clay material, exhibits remarkable effect on wound hemostasis, yet its efficacy is critically limited by heterogeneous particle sizes. Therefore, sorting based on size differences is essential to improve its performance. However, kaolinite with a layer structure presents challenges in sorting compared to spherical or elliptical materials, and the size distribution ranges continuously from nanometers to micrometers, which poses significant challenges for precise sorting. Hence, we developed a dual-layer microfluidic filtration chip, to enable high-throughput sorting of kaolinite micro/nanoflakes (size from 1.582 to 0.377 μm). The dual-layer filter membrane structure with graded pore sizes enabled selective sorting of kaolinite particles within a specific size range, and the co-flow fluid arrangement was employed to alleviate membrane clogging. The hemostatic properties of kaolinite particles with different sizes were evaluated through <em>in vivo</em> and <em>in vitro</em> experiments, revealing the significant size-dependent effects of kaolinite on wound hemostasis. The mechanism of different sizes of kaolinite in the process of coagulation, especially the effect on platelet activation and coagulation factor activation, provided a theoretical basis for optimizing kaolinite-based hemostatic materials. This work established a scalable microfluidic strategy for precise sorting of sheet nanomaterials and improved the translational potential of kaolinite in emergency wound hemostasis.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 14","pages":" 3570-3580"},"PeriodicalIF":6.1,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2025/lc/d5lc00274e?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144228826","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A three-dimensional microfluidic device embedded within a thermal cycler tube for electrokinetic DNA extraction.","authors":"Qi Jiang, Xuehao Zang, Yilu Wang, Alexandre S Avaro, Diego A Huyke, Juan G Santiago","doi":"10.1039/d4lc01062k","DOIUrl":"https://doi.org/10.1039/d4lc01062k","url":null,"abstract":"<p><p>Microfluidic devices have been widely used in modern chemical and biological analyses as stand-alone units, typically in series with other equipment such as extraction columns, manual or robotic pipetting, and even advanced next-generation sequencing systems. While microfluidic devices have enhanced various aspects of laboratory workflows, their integration with established commercial assay platforms remains limited. To this end, we developed a three-dimensional microfluidic insert embedded directly into a commercially available polymerase chain reaction (PCR) tube. This integration creates a microfluidic device compatible with conventional thermal cyclers, which support complex temperature cycling and multiplexed fluorescence detection. The integrated system facilitates key bioassay functions like nucleic acid purification through a selective ionic focusing method known as isotachophoresis (ITP), PCR amplification, and real-time fluorescence detection. We validated the performance of the integrated system by purifying nucleic acids from raw human serum samples and detecting exogenous SARS-CoV-2 N gene using FAM-labeled TaqMan probes, with both the DNA extraction and detection carried out within the same PCR tube. We achieved a detection sensitivity of 100 cp μL^-1 within a total process time of 60 min in these experiments. Human serum samples processed without purification show no PCR amplification results. This integrated system demonstrates the powerful concept of integrating microfluidic structures into form factors compatible with the highly complex and sensitive operation of current off-the-shelf systems.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144223771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-throughput enrichment of functional disulfide-rich peptides by droplet microfluidics†","authors":"Jing Xie, Kuok Yap, Simon J. de Veer, Selvakumar Edwardraja, Thomas Durek, Matt Trau, David J. Craik and Conan K. Wang","doi":"10.1039/D5LC00261C","DOIUrl":"10.1039/D5LC00261C","url":null,"abstract":"<p >Disulfide-rich peptides (DRPs) have evolved intricate topologies to carry out a wide range of bioactivities throughout nature, <em>e.g.</em>, in fungi, insects, plants and animals, and have proven applications in medicine and agriculture. To discover novel DRPs, it is now routine to screen DRP libraries for target affinity, but target binding does not necessarily correlate with function. This study reports an innovative platform for screening of DRP libraries based on the functional endpoint of biochemical reactions within picoliter-sized water-in-oil droplets. We leveraged yeast secretory expression to ensure proper assembly of disulfide connectivity, and thus peptide shape, and engineered customizable strains for facile detection of function (<em>i.e.</em>, protease inhibitory activity) for libraries of DRPs. Rapid enrichment of a potent trypsin inhibitor (MCoTI-II) from a &gt;100 000 pool of randomized variants across four rounds of selection was achieved, far exceeding the library sizes explored previously for peptide systems in droplet microfluidics. This developed platform provides a foundation to explore the functional engineering of DRPs.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 14","pages":" 3525-3536"},"PeriodicalIF":6.1,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144211173","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Patient-derived organotypic tumor spheroids, tumoroids, and organoids: advancing immunotherapy using state-of-the-art 3D tumor model systems","authors":"David J. Bozym, David X. Zheng, Or-Yam Revach, Amir Aref and Russell W. Jenkins","doi":"10.1039/D5LC00062A","DOIUrl":"10.1039/D5LC00062A","url":null,"abstract":"<p >Preclinical <em>ex vivo</em> models capable of probing patient-specific tumor–immune interactions are particularly attractive candidates for interrogating mechanisms of resistance, developing predictors of response as well as assessing next-generation immunotherapeutics. By maintaining features of a patient's own tumor microenvironment, such patient-derived <em>ex vivo</em> models are poised to meaningfully contribute to the functional assessment of individual tumors to provide a tailored approach to treatment. Among contemporary <em>ex vivo</em> models, patient-derived organotypic tumor spheroids (PDOTS) have emerged as a promising microfluidic-based platform that is well positioned to become a useful tool for precision medicine efforts. The advantages and limitations of PDOTS and related state-of-the-art patient-derived tumor models, as well as ongoing challenges facing the clinical implementation of patient-derived <em>ex vivo</em> tumor models, are reviewed.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 13","pages":" 3038-3059"},"PeriodicalIF":6.1,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144211177","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Controllable pump-free electrokinetic-driven microdevice for single-cell electrorotation†","authors":"Jianming Shu, Xijiang Wang and Liang Huang","doi":"10.1039/D5LC00202H","DOIUrl":"10.1039/D5LC00202H","url":null,"abstract":"<p >Single-cell electrorotation (ROT) has emerged as a fundamental technique for characterizing cellular electrical properties, yet conventional methodologies face significant limitations including laborious cell loading procedures, time-consuming measurements, low throughput, and confined effective operational regions. To address these challenges, we present an innovative pump-free single-cell ROT device that synergistically integrates electroosmotic flow (EOF) with ROT technologies. Our design employs time-division multiplexed electrical signal modulation to achieve real-time regulation of EOF velocity and directionality, effectively resolving cell positioning challenges while eliminating the need for complex pumping system. This approach not only reduces experimental cost but also significantly simplifies operational complexity. Furthermore, the implementation of thick-electrode architecture successfully mitigates electric field spatial attenuation, thereby expanding the effective ROT zone and enhancing measurement stability and precision. The capability of the proposed method was tested by rotating yeast and colon cancer cells. Using this device, we quantified the membrane permittivity and cytoplasmic conductivity of these two cell types, revealing differences in the electrical parameters of different types of cells. We envision that the pump-free single-cell ROT microdevice will provide a new platform for convenient and high-throughput cell electrical characterization.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 14","pages":" 3516-3524"},"PeriodicalIF":6.1,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144211175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Parallel dilution microfluidic device for enabling logarithmic concentration generation in molecular diagnostics†","authors":"Akira Miyajima, Fumiya Nishimura, Daigo Natsuhara, Yuka Kiba, Shunya Okamoto, Moeto Nagai, Tadashi Yamamuro, Masashi Kitamura and Takayuki Shibata","doi":"10.1039/D5LC00356C","DOIUrl":"10.1039/D5LC00356C","url":null,"abstract":"<p >In this study, we present a genetic diagnostic device with a four-stepwise logarithmic dilution capability for rapid and reliable detection of target nucleic acids in a single operation using the colorimetric loop-mediated isothermal amplification (LAMP) method. An innovative feature is the confluent point with differing microchannel heights ensuring the synchronized inflow of liquids while preventing backflow, even under large volumetric flow rate variations (10–10 000-fold). This enabled the independent generation of each dilution factor under constant pressure. Furthermore, an integrated asymmetric micromixer effectively mixed two liquids under laminar flow conditions, enabling simultaneous dispensing of the mixed solution at uniform concentrations into five microchambers for each dilution factor. Additionally, a permanent stop valve in the outlet of each microchamber prevented leakages, minimizing the waste of valuable samples and reagents. We demonstrate that diluted samples were accurately prepared at the intended logarithmic dilution factors in a single operation using purified cannabis seed DNA, achieving detection sensitivity similar to that of conventional turbidity-based LAMP assays. Moreover, we used crudely extracted cannabis resin DNA, which contains several gene amplification inhibitors, successfully detecting the target nucleic acids in a single test. Overall, this versatile device eliminates extensive manual sample preparation and has potential for on-site genetic testing in applications such as infectious disease detection, food safety, and illegal drug testing.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 13","pages":" 3242-3253"},"PeriodicalIF":6.1,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2025/lc/d5lc00356c?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144211174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transient osmotic flows in a microfluidic channel: measurements of solute permeability and reflection coefficients of hydrogel membranes†","authors":"Julien Renaudeau, Pierre Lidon and Jean-Baptiste Salmon","doi":"10.1039/D5LC00276A","DOIUrl":"10.1039/D5LC00276A","url":null,"abstract":"<p >We first highlight theoretically a microfluidic configuration that allows to measure two fundamental parameters describing mass transport through a membrane: the solute permeability coefficient <img><small>_D</small>, and the associated reflection coefficient <em>σ</em>. This configuration exploits the high confinement of microfluidic geometries to relate these two coefficients to the dynamics of a transient flow induced by forward osmosis through a membrane embedded in a chip. We then applied this methodology to hydrogel membranes photo-crosslinked in a microchannel with <em>in situ</em> measurements of osmotically-induced flows. These experiments enable us to estimate <img><small>_D</small> and <em>σ</em> and their dependence on the molecular weight of the solute under consideration, ultimately leading to a precise estimate of the molecular weight cut-off of these hydrogel membranes.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 14","pages":" 3549-3558"},"PeriodicalIF":6.1,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144219134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}