Hyun-Kyung Woo, Sangjin Seo, Advitiya Mahajan, Seoyoung Lee, Seoyoon Bae, Jeremy Quintana, Changhyun Kim, Alptekin Aksan, Hakho Lee
Storage options for extracellular vesicles (EVs) remain limited, constraining their clinical and research applications. Conventional low-temperature freezing (-80 °C), although the established standard, requires substantial resources and presents logistical challenges. Here, we report an AridEx (Ambient Retention in Disc of Extracellular vesicles) approach for efficient room-temperature storage and recovery of EVs. AridEx leverages isothermal vitrification principles to preserve EVs, utilizing an electrospun trehalose-dextranol matrix as a xeroprotectant. The xeroprotectant matrix, combined with highly gas-permeable membranes, was integrated into a centrifugal microfluidic device, enabling the rapid desiccation of EV samples under mild vacuum conditions. The dried samples were stored directly on the disc at ambient temperature and subsequently recovered on demand via on-disc rehydration and EV isolation. AridEx achieved preservation efficacy equivalent to that at -80 °C, preserving EV particle counts and surface proteins during ambient storage periods of at least 6 months. In a pilot clinical study, AridEx-preserved plasma retained consistent biomarker signals for accurate cancer diagnosis. This compact and cartridge-based platform is manufacturable and could offer a sustainable alternative to low-temperature sample storage for diagnostic applications.
{"title":"Rapid Desiccation and On-disc Rehydration of Extracellular Vesicles for Non-cryogenic Preservation","authors":"Hyun-Kyung Woo, Sangjin Seo, Advitiya Mahajan, Seoyoung Lee, Seoyoon Bae, Jeremy Quintana, Changhyun Kim, Alptekin Aksan, Hakho Lee","doi":"10.1039/d6lc00014b","DOIUrl":"https://doi.org/10.1039/d6lc00014b","url":null,"abstract":"Storage options for extracellular vesicles (EVs) remain limited, constraining their clinical and research applications. Conventional low-temperature freezing (-80 °C), although the established standard, requires substantial resources and presents logistical challenges. Here, we report an AridEx (Ambient Retention in Disc of Extracellular vesicles) approach for efficient room-temperature storage and recovery of EVs. AridEx leverages isothermal vitrification principles to preserve EVs, utilizing an electrospun trehalose-dextranol matrix as a xeroprotectant. The xeroprotectant matrix, combined with highly gas-permeable membranes, was integrated into a centrifugal microfluidic device, enabling the rapid desiccation of EV samples under mild vacuum conditions. The dried samples were stored directly on the disc at ambient temperature and subsequently recovered on demand via on-disc rehydration and EV isolation. AridEx achieved preservation efficacy equivalent to that at -80 °C, preserving EV particle counts and surface proteins during ambient storage periods of at least 6 months. In a pilot clinical study, AridEx-preserved plasma retained consistent biomarker signals for accurate cancer diagnosis. This compact and cartridge-based platform is manufacturable and could offer a sustainable alternative to low-temperature sample storage for diagnostic applications.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"18 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147507635","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}
Benedetta Marmiroli, Sumea Klokic, Barbara Sartori, Marie Reißenbüchel, Alessio Turchet, Heinz Amenitsch
Microfluidic devices are increasingly used in synchrotron-based experiments to deliver and probe liquid samples, offering advantages such as minimal sample consumption and reduced radiation damage. Despite their growing use, few devices have been specifically designed for monitoring liquids under photoexcitation, a promising approach for fast structural transitions. Here, a microfluidic device is presented that is transparent to X-rays in one direction and simultaneously transmits UV and visible light illumination to the sample in the perpendicular direction. The device is fabricated using lamination and UV lithography on a dry-film resist, eliminating the need for cleanroom facilities and simplifying production. Its multi-wavelength transparency was validated through UV–Vis spectroscopy, where photoexcitation at different wavelengths induced reversible trans–to-cis isomerization of azobenzene and fluoro-azobenzene. X-ray transparency was verified through small-angle X-ray scattering (SAXS) measurements on hemoglobin and CO-ligated hemoglobin, both of which are sensitive to quaternary structural changes. These results confirm the suitability of the device for resolving protein structures and detecting subtle conformational changes of the type commonly encountered in photo-induced modulation. Initial proof- of- concept measurements demonstrate the feasibility of temperature-jump (T-jump) experiments, and the same architecture is readily extendable to time resolved pump–probe studies, providing a versatile platform for studying structural evolution in liquid samples using synchrotron SAXS.
{"title":"Multi-Wavelength transparent microfluidic device for UV-Visible illumination and X-ray Scattering studies of Photoactive Systems","authors":"Benedetta Marmiroli, Sumea Klokic, Barbara Sartori, Marie Reißenbüchel, Alessio Turchet, Heinz Amenitsch","doi":"10.1039/d5lc01116g","DOIUrl":"https://doi.org/10.1039/d5lc01116g","url":null,"abstract":"Microfluidic devices are increasingly used in synchrotron-based experiments to deliver and probe liquid samples, offering advantages such as minimal sample consumption and reduced radiation damage. Despite their growing use, few devices have been specifically designed for monitoring liquids under photoexcitation, a promising approach for fast structural transitions. Here, a microfluidic device is presented that is transparent to X-rays in one direction and simultaneously transmits UV and visible light illumination to the sample in the perpendicular direction. The device is fabricated using lamination and UV lithography on a dry-film resist, eliminating the need for cleanroom facilities and simplifying production. Its multi-wavelength transparency was validated through UV–Vis spectroscopy, where photoexcitation at different wavelengths induced reversible trans–to-cis isomerization of azobenzene and fluoro-azobenzene. X-ray transparency was verified through small-angle X-ray scattering (SAXS) measurements on hemoglobin and CO-ligated hemoglobin, both of which are sensitive to quaternary structural changes. These results confirm the suitability of the device for resolving protein structures and detecting subtle conformational changes of the type commonly encountered in photo-induced modulation. Initial proof- of- concept measurements demonstrate the feasibility of temperature-jump (T-jump) experiments, and the same architecture is readily extendable to time resolved pump–probe studies, providing a versatile platform for studying structural evolution in liquid samples using synchrotron SAXS.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"16 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147506964","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}
The precise enrichment and size-selective separation of sub-100 nm biological nanoparticles such as exosomes remain challenging due to their small size and heterogeneity. Conventional GHz acoustofluidic systems suffer from unstable acoustic streaming vortices at channel entrances, leading to particle leakage and limited trapping efficiency. Here, we present a tunable squeeze-activated GHz acoustofluidics (TSGA) platform that overcomes these limitations through dynamic and symmetric deformation of the microchannel. By adjusting pneumatic pressure and acoustic power in real time, the system enables on-demand modulation of the acoustic streaming field and particle migration paths. The squeeze-induced arched profile guides nanoparticles toward stable vortex regions while enhancing their exposure frequency to high-gradient acoustic radiation force. This programmable mechanism allows continuous enrichment of particles down to 50 nm, achieving a single-round enrichment factor of 2.38 for 150 nm particles with >75% recovery efficiency. Moreover, through coordinated pressure-acoustic tuning, a multistage separation strategy successfully isolates 77 nm particles from complex mixtures, increasing purity from 30.3% to 80.6%, and purifies exosome subpopulations with high resolution. The TSGA platform provides a robust, label-free, and dynamically tunable approach for scalable nanoscale bioparticle processing, promising advances in exosome research and liquid biopsy diagnostics.
{"title":"Tunable squeeze-activated GHz acoustofluidics for stable trapping and separation of sub-100 nm nanoparticles.","authors":"Yiming Liu,Wei Wei,Hang Qi,Shuaihua Zhang,Yongqi Chen,Yaping Wang,Xuexin Duan","doi":"10.1039/d6lc00106h","DOIUrl":"https://doi.org/10.1039/d6lc00106h","url":null,"abstract":"The precise enrichment and size-selective separation of sub-100 nm biological nanoparticles such as exosomes remain challenging due to their small size and heterogeneity. Conventional GHz acoustofluidic systems suffer from unstable acoustic streaming vortices at channel entrances, leading to particle leakage and limited trapping efficiency. Here, we present a tunable squeeze-activated GHz acoustofluidics (TSGA) platform that overcomes these limitations through dynamic and symmetric deformation of the microchannel. By adjusting pneumatic pressure and acoustic power in real time, the system enables on-demand modulation of the acoustic streaming field and particle migration paths. The squeeze-induced arched profile guides nanoparticles toward stable vortex regions while enhancing their exposure frequency to high-gradient acoustic radiation force. This programmable mechanism allows continuous enrichment of particles down to 50 nm, achieving a single-round enrichment factor of 2.38 for 150 nm particles with >75% recovery efficiency. Moreover, through coordinated pressure-acoustic tuning, a multistage separation strategy successfully isolates 77 nm particles from complex mixtures, increasing purity from 30.3% to 80.6%, and purifies exosome subpopulations with high resolution. The TSGA platform provides a robust, label-free, and dynamically tunable approach for scalable nanoscale bioparticle processing, promising advances in exosome research and liquid biopsy diagnostics.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"19 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147502434","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}
Ana Mesic, Antonietta Messina, Zoe Tiprez, Safa Ismail, Nicolas Huang, Benoit Charlot, Sakina Bensalem, Jean-Charles Duclos-Vallee, Bruno Le Pioufle
The limited predictive power of animal models remains a major bottleneck in drug development, particularly in assessing drug-induced liver injury (DILI). To address this, we developed a novel in vitro liver-on-a-chip platform focused on modeling the space of Disse (sD)-a critical yet underrepresented microenvironment mediating endothelial-hepatic interactions. The system integrates a biocompatible sodium alginate hydrogel whose mechanical properties were optimized to mimic physiological liver stiffness, enabling molecular cross-talk between human liver sinusoidal endothelial cells (LSECs) and HepaRG hepatocytes without direct contact. Under dynamic perfusion, the co-culture maintained viability and polarization for eight days, forming organized tissues with functional bile canaliculi. The presence of LSECs markedly enhanced hepatic performance, reflected by increased albumin and urea secretion and activation of proregenerative secretory pathways. Proof-of-concept studies with chronic acetaminophen exposure demonstrated the model's capacity to reproduce hepatotoxic responses, confirming its predictive relevance. This versatile and physiologically relevant microphysiological platform offers a powerful tool for studying endothelial-hepatic communication, modeling liver pathologies, and improving preclinical DILI testing. Its modular design enables future integration of Kupffer and stellate cells to simulate immune and fibrotic responses, extending its applicability to complex liver disease modeling.
{"title":"In Vitro Space of Disse Model for Exploration of Drug Induced Hepatotoxicity","authors":"Ana Mesic, Antonietta Messina, Zoe Tiprez, Safa Ismail, Nicolas Huang, Benoit Charlot, Sakina Bensalem, Jean-Charles Duclos-Vallee, Bruno Le Pioufle","doi":"10.1039/d5lc01139f","DOIUrl":"https://doi.org/10.1039/d5lc01139f","url":null,"abstract":"The limited predictive power of animal models remains a major bottleneck in drug development, particularly in assessing drug-induced liver injury (DILI). To address this, we developed a novel in vitro liver-on-a-chip platform focused on modeling the space of Disse (sD)-a critical yet underrepresented microenvironment mediating endothelial-hepatic interactions. The system integrates a biocompatible sodium alginate hydrogel whose mechanical properties were optimized to mimic physiological liver stiffness, enabling molecular cross-talk between human liver sinusoidal endothelial cells (LSECs) and HepaRG hepatocytes without direct contact. Under dynamic perfusion, the co-culture maintained viability and polarization for eight days, forming organized tissues with functional bile canaliculi. The presence of LSECs markedly enhanced hepatic performance, reflected by increased albumin and urea secretion and activation of proregenerative secretory pathways. Proof-of-concept studies with chronic acetaminophen exposure demonstrated the model's capacity to reproduce hepatotoxic responses, confirming its predictive relevance. This versatile and physiologically relevant microphysiological platform offers a powerful tool for studying endothelial-hepatic communication, modeling liver pathologies, and improving preclinical DILI testing. Its modular design enables future integration of Kupffer and stellate cells to simulate immune and fibrotic responses, extending its applicability to complex liver disease modeling.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"44 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147492598","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}
Francesca Fanizza, Simone Perottoni, Lucia Boeri, Francesca Donnaloja, Francesca Negro, Francesca Pugli, Gianluigi Forloni, Carmen Giordano, Diego Albani
Correction for 'A gut-brain axis on-a-chip platform for drug testing challenged with donepezil' by Francesca Fanizza et al., Lab Chip, 2025, 25, 1854-1874, https://doi.org/10.1039/D4LC00273C.
{"title":"Correction: A gut-brain axis on-a-chip platform for drug testing challenged with donepezil.","authors":"Francesca Fanizza, Simone Perottoni, Lucia Boeri, Francesca Donnaloja, Francesca Negro, Francesca Pugli, Gianluigi Forloni, Carmen Giordano, Diego Albani","doi":"10.1039/d5lc90130h","DOIUrl":"https://doi.org/10.1039/d5lc90130h","url":null,"abstract":"<p><p>Correction for 'A gut-brain axis on-a-chip platform for drug testing challenged with donepezil' by Francesca Fanizza <i>et al.</i>, <i>Lab Chip</i>, 2025, <b>25</b>, 1854-1874, https://doi.org/10.1039/D4LC00273C.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147483905","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}
Hyo-il Jung, Zhaoyu Zhang, Jaejeung Kim, Jinwoo Hwang, Hyunjo Seo, Geonha Kim, Seoyeon Choi, Kyung-A Hyun
Lipid nanoparticles (LNPs) are a key platform for nucleic acid delivery, but their industrial-scale production remains a critical bottleneck. While microfluidics ensures high-quality LNP production, conventional single layer strategies suffer from low throughput. Existing scaling strategies, like parallel or 3D devices, face limitations in flow distribution and fabrication complexity, failing to balance throughput with LNP quality. Here, we report a robust, vertically stacked microfluidic cartridge (VeSMiC) fabricated using polycarbonate (PC) that successfully addresses the flow distribution challenge. This platform integrates a hydrodynamic tapered inlet structure, ensuring uniform flow partitioning across layers, with re-Tesla mixers for rapid synthesis. A five-layer device was scalable fabricated using a two-step surface modification and a robust bonding method. This device enabled stable operation at a high total flow rate (0.96 L/hour) operation and increased LNP throughput 7-fold compared to single channel. Critically, this high throughput did not compromise quality, LNPs consistently maintained an average size of 100-150 nm, a low polydispersity index (PDI) below 0.12, and high encapsulation efficiency above 96%. Furthermore, they also demonstrated significant therapeutic efficacy in an in vitro wound model. Notably, an 80-cartridge platform is projected to achieve a production flow rate of approximately 80 L/hour, validating this platform as a viable solution for industrial-scale LNP manufacturing.
{"title":"Vertical numbering-up microfluidic architecture for scalable and homogeneous lipid nanoparticle production","authors":"Hyo-il Jung, Zhaoyu Zhang, Jaejeung Kim, Jinwoo Hwang, Hyunjo Seo, Geonha Kim, Seoyeon Choi, Kyung-A Hyun","doi":"10.1039/d5lc01130b","DOIUrl":"https://doi.org/10.1039/d5lc01130b","url":null,"abstract":"Lipid nanoparticles (LNPs) are a key platform for nucleic acid delivery, but their industrial-scale production remains a critical bottleneck. While microfluidics ensures high-quality LNP production, conventional single layer strategies suffer from low throughput. Existing scaling strategies, like parallel or 3D devices, face limitations in flow distribution and fabrication complexity, failing to balance throughput with LNP quality. Here, we report a robust, vertically stacked microfluidic cartridge (VeSMiC) fabricated using polycarbonate (PC) that successfully addresses the flow distribution challenge. This platform integrates a hydrodynamic tapered inlet structure, ensuring uniform flow partitioning across layers, with re-Tesla mixers for rapid synthesis. A five-layer device was scalable fabricated using a two-step surface modification and a robust bonding method. This device enabled stable operation at a high total flow rate (0.96 L/hour) operation and increased LNP throughput 7-fold compared to single channel. Critically, this high throughput did not compromise quality, LNPs consistently maintained an average size of 100-150 nm, a low polydispersity index (PDI) below 0.12, and high encapsulation efficiency above 96%. Furthermore, they also demonstrated significant therapeutic efficacy in an in vitro wound model. Notably, an 80-cartridge platform is projected to achieve a production flow rate of approximately 80 L/hour, validating this platform as a viable solution for industrial-scale LNP manufacturing.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"17 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147492599","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}
Continuous microscale purification requires analytical methods that provide deterministic fluid handling, precise temporal control, and contamination-free fraction discrimination. Existing microfluidic and benchtop chromatography systems only partially address these needs, leaving a gap for methods that support tightly coordinated, programmable purification cycles. This work presents a microfluidic continuous protein purification method that uses digitally programmable inlet (ICV) and collection (CCV) valves to establish a logic-driven chromatography operation. Sub-second buffer switching and deterministic routing across parallel affinity columns enable a reproducible and algorithm-defined purification sequence. Temporal gating through the CCV provides real-time, profile-guided fraction selection that isolates high-concentration eluates while effectively removing tailing segments. Using GFP-His6 as a model substrate, the system maintains 70–89% purity over ten uninterrupted cycles, demonstrating strong cycle-to-cycle stability. Purification of His6-tagged TRAIL further confirms compatibility with structurally sensitive biologics and preservation of functional activity. The compact, modular, and single-use architecture minimizes dead volume, prevents cross-contamination, and accommodates diverse chromatographic modes. By combining programmable valve logic with time-resolved elution control, this work advances microfluidic platforms from diagnostic tools toward autonomous and precision-controlled process operations. The method provides a broadly applicable analytical framework for microscale purification and supports the development of next-generation bioseparation and continuous biomanufacturing technologies.
{"title":"Digitally Programmable Microfluidic Valving for Autonomous, High-Resolution Continuous Chromatographic Purification","authors":"Yi-Cheng Liao, Chih-Yi Huang, Yu-Chuan Tang, Cheng-Hsian Wu, Yu-Hsuan Chi, I-Wei Chen, Ya-Hui Lin, Hsuan‐Yu Mu, Yunching Chen, Fu-Fei Hsu, Jen-Huang Huang","doi":"10.1039/d6lc00015k","DOIUrl":"https://doi.org/10.1039/d6lc00015k","url":null,"abstract":"Continuous microscale purification requires analytical methods that provide deterministic fluid handling, precise temporal control, and contamination-free fraction discrimination. Existing microfluidic and benchtop chromatography systems only partially address these needs, leaving a gap for methods that support tightly coordinated, programmable purification cycles. This work presents a microfluidic continuous protein purification method that uses digitally programmable inlet (ICV) and collection (CCV) valves to establish a logic-driven chromatography operation. Sub-second buffer switching and deterministic routing across parallel affinity columns enable a reproducible and algorithm-defined purification sequence. Temporal gating through the CCV provides real-time, profile-guided fraction selection that isolates high-concentration eluates while effectively removing tailing segments. Using GFP-His6 as a model substrate, the system maintains 70–89% purity over ten uninterrupted cycles, demonstrating strong cycle-to-cycle stability. Purification of His6-tagged TRAIL further confirms compatibility with structurally sensitive biologics and preservation of functional activity. The compact, modular, and single-use architecture minimizes dead volume, prevents cross-contamination, and accommodates diverse chromatographic modes. By combining programmable valve logic with time-resolved elution control, this work advances microfluidic platforms from diagnostic tools toward autonomous and precision-controlled process operations. The method provides a broadly applicable analytical framework for microscale purification and supports the development of next-generation bioseparation and continuous biomanufacturing technologies.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"60 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147478676","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}
Fariba Malekpour Galogahi, Haotian Cha, Sharda Yadav, Hang Thu Ta, Nam-Trung Nguyen
The ability to sort and separate large cellular subpopulations based on their intrinsic properties underpins a wide range of biological, diagnostic, and therapeutic applications. For many of these applications, maintaining cellular homogeneity within the confinement of a droplet is critical for accurate quantitative analysis and subsequent processing. Although microfluidic platforms have successfully enabled the separation of cellular subpopulations from heterogeneous samples, the lack of droplet-based encapsulation post separation remains a major bottle neck for achieving high-throughput single-cell analysis.Here, we address this limitation by developing an integrated microfluidic device that enables size-based cell separation and simultaneously encapsulating single cells into picolitre droplets. The device overcomes unstable encapsulation of cells by uniformly spacing cells prior to the encapsulation process. Proof-of-concept experiments achieved a single-particle encapsulation efficiency of 60% for 15 µm polystyrene beads, exceeding the Poisson limit of ~35% single occupancy. Size-based separation of 15-µm particles from 10-µm particles yielded a separation efficiency of 94.39%, with nearly 60% of the separated particles successfully encapsulated as single particles in droplets. Validation experiments using MDA-MB-231 cancer cells dispersed in white blood cells (WBCs) demonstrated a 92.74% separation efficiency, with approximately 28% of cancer cells encapsulated as single cells within droplets.
{"title":"Integrated Microfluidic Platform for Inertial Separation and Encapsulation of Single Cells in Droplets","authors":"Fariba Malekpour Galogahi, Haotian Cha, Sharda Yadav, Hang Thu Ta, Nam-Trung Nguyen","doi":"10.1039/d6lc00085a","DOIUrl":"https://doi.org/10.1039/d6lc00085a","url":null,"abstract":"The ability to sort and separate large cellular subpopulations based on their intrinsic properties underpins a wide range of biological, diagnostic, and therapeutic applications. For many of these applications, maintaining cellular homogeneity within the confinement of a droplet is critical for accurate quantitative analysis and subsequent processing. Although microfluidic platforms have successfully enabled the separation of cellular subpopulations from heterogeneous samples, the lack of droplet-based encapsulation post separation remains a major bottle neck for achieving high-throughput single-cell analysis.Here, we address this limitation by developing an integrated microfluidic device that enables size-based cell separation and simultaneously encapsulating single cells into picolitre droplets. The device overcomes unstable encapsulation of cells by uniformly spacing cells prior to the encapsulation process. Proof-of-concept experiments achieved a single-particle encapsulation efficiency of 60% for 15 µm polystyrene beads, exceeding the Poisson limit of ~35% single occupancy. Size-based separation of 15-µm particles from 10-µm particles yielded a separation efficiency of 94.39%, with nearly 60% of the separated particles successfully encapsulated as single particles in droplets. Validation experiments using MDA-MB-231 cancer cells dispersed in white blood cells (WBCs) demonstrated a 92.74% separation efficiency, with approximately 28% of cancer cells encapsulated as single cells within droplets.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"6 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147478678","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}
Efficient micromixing in enclosed microchannels is essential for reliable lab-on-a-chip operation but typically requires planar or multilayer soft-lithographic fabrication, limiting geometric freedom and increasing production complexity. Here, we report a fully monolithic split-and-recombine (SAR) micromixer fabricated in a single step using stereolithography digital light processing (SLA-DLP), eliminating molds, bonding, and cleanroom processing. The three-dimensional SAR architecture systematically divides, reorients, and recombines fluid streams, enabling high mixing performance over a wide range of operating conditions. Computational fluid dynamics simulations and experimental validation show excellent agreement, achieving mixing efficiencies above 0.90 across Reynolds numbers from 0.1 to 100. Leveraging its compact and robust performance, the micromixer was integrated into a five-output microfluidic con-centration gradient generator, which produced stable and reproducible concentration profiles for both fluorescent tracers and protein solutions. The complete device, including microchannels and functional fluidic features, was printed in under 1.5 h using a standard desktop SLA-DLP system. These results demonstrate that additive manufacturing can deliver high-performance micromixing capabilities, establishing a rapid, accessible, and fully digital route for the fabrication of advanced microfluidic systems.
{"title":"Monolithic 3D-Printed Split-and-Recombine Micromixer Integrated into a Microfluidic Concentration Gradient Generator","authors":"Francisco Navarro Molina, J Paliwal, Elham Salimi","doi":"10.1039/d5lc01095k","DOIUrl":"https://doi.org/10.1039/d5lc01095k","url":null,"abstract":"Efficient micromixing in enclosed microchannels is essential for reliable lab-on-a-chip operation but typically requires planar or multilayer soft-lithographic fabrication, limiting geometric freedom and increasing production complexity. Here, we report a fully monolithic split-and-recombine (SAR) micromixer fabricated in a single step using stereolithography digital light processing (SLA-DLP), eliminating molds, bonding, and cleanroom processing. The three-dimensional SAR architecture systematically divides, reorients, and recombines fluid streams, enabling high mixing performance over a wide range of operating conditions. Computational fluid dynamics simulations and experimental validation show excellent agreement, achieving mixing efficiencies above 0.90 across Reynolds numbers from 0.1 to 100. Leveraging its compact and robust performance, the micromixer was integrated into a five-output microfluidic con-centration gradient generator, which produced stable and reproducible concentration profiles for both fluorescent tracers and protein solutions. The complete device, including microchannels and functional fluidic features, was printed in under 1.5 h using a standard desktop SLA-DLP system. These results demonstrate that additive manufacturing can deliver high-performance micromixing capabilities, establishing a rapid, accessible, and fully digital route for the fabrication of advanced microfluidic systems.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"27 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147478679","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}
Carbon dioxide enhanced oil recovery (CO2-EOR) has been recognized as a viable pathway for carbon capture, utilization, and storage (CCUS). Among its variants, miscible CO2-EOR offers a considerable additional oil recovery of approximately 5–20%, making the determination of minimum miscibility pressure (MMP) a critical design consideration. In this study, we employ a high-pressure microfluidic platform to investigate the miscibility transition between CO2 and n-decane at temperatures (T) of 40, 50, 70, and 90 °C. At T = 40 °C, with increasing pressure (P), microfluidic visualization reveals a series of distinct flow regimes: dripping, quasi-steady jetting, unsteady jetting, transitional, and ultimately diffusive regimes. In the diffusive regime, miscibility is achieved through intensive mixing, leading to the disappearance of the fluid–fluid interface. Based on these microfluidic observations, we propose a new criterion for MMP determination: the minimum pressure required to reach the diffusive regime for the dynamic CO2–oil flow. The experimentally determined MMP values show good agreement with previous microfluidic studies and predictions from the Peng–Robinson equation of state (PR-EOS). Furthermore, the MMP increases linearly with temperature from 40 to 90 °C, consistent with the reduced solubility of CO2 in n-decane at higher temperatures. This microfluidic method provides a rapid and visual approach to assess miscibility transitions in CO2-EOR applications.
{"title":"Microfluidic determination of minimum miscibility pressure (MMP) in dynamic CO2/n-decane flow","authors":"Junyi Yang, Peichun Amy Tsai","doi":"10.1039/d5lc00616c","DOIUrl":"https://doi.org/10.1039/d5lc00616c","url":null,"abstract":"Carbon dioxide enhanced oil recovery (CO<small><sub>2</sub></small>-EOR) has been recognized as a viable pathway for carbon capture, utilization, and storage (CCUS). Among its variants, miscible CO<small><sub>2</sub></small>-EOR offers a considerable additional oil recovery of approximately 5–20%, making the determination of minimum miscibility pressure (MMP) a critical design consideration. In this study, we employ a high-pressure microfluidic platform to investigate the miscibility transition between CO<small><sub>2</sub></small> and <em>n</em>-decane at temperatures (<em>T</em>) of 40, 50, 70, and 90 °C. At <em>T</em> = 40 °C, with increasing pressure (<em>P</em>), microfluidic visualization reveals a series of distinct flow regimes: dripping, quasi-steady jetting, unsteady jetting, transitional, and ultimately diffusive regimes. In the diffusive regime, miscibility is achieved through intensive mixing, leading to the disappearance of the fluid–fluid interface. Based on these microfluidic observations, we propose a new criterion for MMP determination: the minimum pressure required to reach the diffusive regime for the dynamic CO<small><sub>2</sub></small>–oil flow. The experimentally determined MMP values show good agreement with previous microfluidic studies and predictions from the Peng–Robinson equation of state (PR-EOS). Furthermore, the MMP increases linearly with temperature from 40 to 90 °C, consistent with the reduced solubility of CO<small><sub>2</sub></small> in <em>n</em>-decane at higher temperatures. This microfluidic method provides a rapid and visual approach to assess miscibility transitions in CO<small><sub>2</sub></small>-EOR applications.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"1 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147478680","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}
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