Lab on a ChipPub Date : 2025-06-05DOI: 10.1039/d5lc00109a
Anthony Mercader, Sung Kwon Cho
{"title":"Acoustic microstreaming and augmentation of gas exchange using an oscillating membrane towards microfluidic artificial lungs.","authors":"Anthony Mercader, Sung Kwon Cho","doi":"10.1039/d5lc00109a","DOIUrl":"https://doi.org/10.1039/d5lc00109a","url":null,"abstract":"<p><p>This paper presents a novel configuration for generating acoustic microstreaming flows at audible frequencies within a microchannel utilizing a pinned oscillating membrane. The characterization and interactions of these acoustic streaming flows with the streamwise flow within the microchannel are investigated, along with their effects on gas exchange augmentation. Advanced characterization methods and computational fluid dynamics simulations show a similar pattern and magnitude in acoustic streaming, providing evidence that this flexural membrane oscillation is the driving mechanism of the time-averaged vortices. This method exhibits potential application to microfluidic artificial lungs, particularly due to the vertical orientation of the resulting mixing, which facilitates an augmentation of gas exchange across the permeable membrane. Furthermore, it eliminates any obstructions in the microchannel and ensures stability, as opposed to other acoustic streaming methods such as sharp edge and oscillating bubble methods. Successful augmentation of gas exchange by up to 3.7× is demonstrated as shown by characterization of CO<sub>2</sub> transferred into the channel. Scaling up of throughput is also demonstrated with a branching design, featuring a multilayer manifold to avoid undesirable interaction of the streaming flow with the channel geometry.</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":"144223772","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}
Lab on a ChipPub Date : 2025-06-04DOI: 10.1039/d5lc00261c
Jing Xie, Kuok Yap, Simon J. de Veer, Selvakumar Edwardraja, Thomas Durek, Matt Trau, David J. Craik, Conan K. Wang
{"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, Conan K. Wang","doi":"10.1039/d5lc00261c","DOIUrl":"https://doi.org/10.1039/d5lc00261c","url":null,"abstract":"Disulfide-rich peptides (DRPs) have evolved intricate topologies to carry out a wide range of bioactivities throughout nature, e.g., 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 (i.e., protease inhibitory activity) for libraries of DRPs. Rapid enrichment of a potent trypsin inhibitor (MCoTI-II) from a >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.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"13 1","pages":""},"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":"Enhancing cancer cell immunocapture on orientation-controlled nanoimprinted microcone arrays in microgap channels.","authors":"Yuhei Saito, Natsumi Shimmyo, Shuhei Aoyama, Rie Utoh, Minoru Seki, Masumi Yamada","doi":"10.1039/d5lc00143a","DOIUrl":"https://doi.org/10.1039/d5lc00143a","url":null,"abstract":"<p><p>Cancer detection through circulating tumor cell (CTC)-based liquid biopsy has been expected to be a new modality for less-invasive, next-generation diagnosis. However, due to limitations such as the complexity of the cell capture devices, as well as the cost and reproducibility of their fabrication and surface functionalization, these methods are not yet practical for clinical use. In this study, we propose a new strategy for affinity-based selective capture of cancer cells using a microfluidic system integrating nanoengineered polymeric structures. Polycarbonate (PC) sheets with closely assembled microcone arrays were prepared using thermal nanoimprint lithography (T-NIL). These structures are suitable for mass production and can strongly absorb antibodies without the need for chemical linkers. Microgap channels incorporating the microcones were formed by simply sandwiching the sheet between two plates, which were highly functional in capturing cancer cells from blood samples. In this study, we clarified the effect of the orientation angle of the microcone array in a closely packed hexagonal pattern to ensure high capture efficiency even under high flow-rate conditions. The feasibility of detecting cancer cells through post-capture processing was also demonstrated. The cell capture structures proposed in this study are simple and reproducible in their fabrication, highly productive, and practical, and may become a new tool for cell-based cancer diagnosis.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144214402","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}
Lab on a ChipPub Date : 2025-06-04DOI: 10.1039/D5LC00062A
David J. Bozym, David X. Zheng, Or-Yam Revach, Amir Aref and Russell W. Jenkins
{"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}
Lab on a ChipPub Date : 2025-06-04DOI: 10.1039/d5lc00202h
Jianming Shu, Xijiang Wang, Liang Huang
{"title":"Controllable Pump-Free Electrokinetic-Driven Microdevice for Single-Cell Electrorotation","authors":"Jianming Shu, Xijiang Wang, Liang Huang","doi":"10.1039/d5lc00202h","DOIUrl":"https://doi.org/10.1039/d5lc00202h","url":null,"abstract":"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 electrorotation 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. Comprehensive numerical simulations and experimental validations demonstrate the capability of microdevice for accurate determination of cellular parameters, (eg. membrane permittivity εmem and cytoplasmic conductivity σcyto). We envision that the pump-free single-cell ROT microdevice will provide a new platform for convenient and high-throughput cell electrical characterization.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"16 1","pages":""},"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}
Lab on a ChipPub Date : 2025-06-04DOI: 10.1039/D5LC00356C
Akira Miyajima, Fumiya Nishimura, Daigo Natsuhara, Yuka Kiba, Shunya Okamoto, Moeto Nagai, Tadashi Yamamuro, Masashi Kitamura and Takayuki Shibata
{"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}
Lab on a ChipPub Date : 2025-06-03DOI: 10.1039/d5lc00276a
Julien Renaudeau, Pierre Lidon, Jean-Baptiste Salmon
{"title":"Transient osmotic flows in a microfluidic channel: measurements of solute permeability and reflection coefficients of hydrogel membranes","authors":"Julien Renaudeau, Pierre Lidon, Jean-Baptiste Salmon","doi":"10.1039/d5lc00276a","DOIUrl":"https://doi.org/10.1039/d5lc00276a","url":null,"abstract":"We first highlight theoretically a microfluidic configuration that allows to measure two fundamental parameters describing mass transport through a membrane: the solute permeability coefficient L<small><sub>D</sub></small>, and the associated reflection coefficient σ. 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 textit{in situ} measurements of osmotically-induced flows. These experiments enable us to estimateL<small><sub>D</sub></small> and σ 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.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"359 1","pages":""},"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}
{"title":"Ion concentration polarization focusing at a millimeter-scale microbead junction: towards higher volumetric throughput","authors":"Umesha Peramune, Zisun Ahmed, Robbyn Kimberly Anand","doi":"10.1039/d5lc00183h","DOIUrl":"https://doi.org/10.1039/d5lc00183h","url":null,"abstract":"Ion concentration polarization focusing (ICPF) is an electrokinetic technique that has shown promise in achieving even billion-fold preconcentration factors. However, increasing the volumetric throughput of ICPF is challenging because disruptive processes that reduce preconcentration efficiency worsen as the channel cross-section extends beyond the microscale. We previously introduced an approach for mitigating the above challenges in a microfluidic regime. However, in that system, the flow rate was limited to less than 1.0 µL/min. Herein, we report a high throughput and scalable ICPF of charged analytes in a millimeter-scale channel. Using 3D-printed channels of 4.0 mm2 cross-section, we achieve preconcentration factors above 200 fold within 10 min at a flow rate of 30 µL/min. In this system, ICP is accomplished by ion permselective transport through a packed bed of commercially available cation exchange microbeads (30 µm and 200 µm). We investigate the scalability of the approach by comparing the ICPF performance of channels with four distinct cross-sectional areas. While ICPF occurs in all four cases, the degree of preconcentration drops below 100 fold (per 10 min) in channels with cross-sections beyond 4.0 mm2. This drop in efficiency is attributed to dispersion associated with Joule heating. Therefore, by improving the device design to dissipate heat more effectively, we anticipate that this approach can be scaled up further for applications that demand high volumetric throughput electrokinetic focusing.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"5 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144211178","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}
Lab on a ChipPub Date : 2025-05-31DOI: 10.1039/d5lc00340g
Johnson Q Cui, Ruyuan Song, Weihong Song, Ouyang Li, Xin Yuan, Hongbo Zhou, Lu Zhang, Shuhuai Yao
{"title":"High-throughput Monoclonal Antibody Screening from Immunized Rabbits via Droplet Microfluidics","authors":"Johnson Q Cui, Ruyuan Song, Weihong Song, Ouyang Li, Xin Yuan, Hongbo Zhou, Lu Zhang, Shuhuai Yao","doi":"10.1039/d5lc00340g","DOIUrl":"https://doi.org/10.1039/d5lc00340g","url":null,"abstract":"The discovery of monoclonal antibodies (mAbs) is critical to advancing therapeutics, diagnostics, and biomedical research. While mouse-derived mAbs dominate current applications, their limitations—short serum half-life, human immunogenicity, and restricted recognition of human-specific antigens—highlight the need for alternative sources. Rabbit-derived mAbs have been gaining significant traction with their superior antigen-binding affinity, broader epitope diversity, and higher yield potential. However, the absence of well-defined surface markers on rabbit B cells has hindered efficient enrichment strategies, limiting the exploration of this valuable antibody repertoire. In this study, we present an integrated workflow that combines magnetic negative selection with high-throughput droplet microfluidics to overcome these barriers. By optimizing a pan B cell enrichment protocol using a tailored antibody cocktail, we achieved a notable boost in IgG secretion and B cell enrichment. Through two complementary droplet-encapsulated assays using particle aggregation for soluble antigens and reporter cells for membrane-bound antigens, we identified target cells capable of secreting high-affinity IgGs. Subsequent sequencing, in vitro antibody production and characterization confirmed the high affinity rate of the discovered antibodies, outperforming rates previously reported. The use of droplet microfluidics streamlines the analysis of rabbit IgG repertoires, providing a robust platform for rabbit single B cell antibody discovery with promising applications in precision medicines and diagnostics.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"53 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144211179","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":"<i>In situ</i> electrochemical measurement of alkaline phosphatase activity in engineered gut models using a porous membrane electrode device.","authors":"Yoshinobu Utagawa, Takeo Miyake, Yasuhiko Shinoda, Masateru Yamazaki, Hiroya Abe, Hitoshi Shiku, Kosuke Ino","doi":"10.1039/d5lc00341e","DOIUrl":"https://doi.org/10.1039/d5lc00341e","url":null,"abstract":"<p><p><i>In vitro</i> assays utilizing human cells have attracted attention as alternatives to animal testing for drug screening. Microphysiological systems (MPS) have been proposed as cell culture platforms that more closely mimic the <i>in vivo</i> microenvironment. Engineered gut models have been extensively studied using human colorectal adenocarcinoma cell lines (Caco-2 cells). Alkaline phosphatase (ALP) activity indicates Caco-2 cell differentiation and is a component of the chemical barrier that maintains intestinal barrier function. However, conventional absorbance-based methods for ALP measurement can harm cells owing to prolonged exposure to alkaline conditions (approximately 60 min). In this study, we developed an engineered gut model with a porous membrane electrode for <i>in situ</i> measurement of ALP activity. The electrochemical sensors, positioned directly beneath the cells on porous membrane electrodes, allowed for rapid short-term measurements. ALP activity was detected after a 3 min treatment through <i>in situ</i> electrochemical measurements, which indicated low cytotoxicity. Therefore, we measured ALP activity multiple times using the same device over 21 days. The proposed system offers potential for <i>in situ</i> measurement and real-time monitoring in <i>in vitro</i> organ models, including MPS devices.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144179729","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}