Aniruddha Paul, Eric R. Safai, Laura E. de Heus, Anke R. Vollertsen, Kevin Weijgertse, Bjorn de Wagenaar, Hossein E. Amirabadi, Evita van de Steeg, Mathieu Odijk, Andries D. van der Meer and Joshua Loessberg-Zahl
Organ-on-chips (OoC) have the potential to revolutionize drug testing. However, the fragmented landscape of existing OoC systems leads to wasted resources and collaboration barriers, slowing broader adoption. To unite the ecosystem, there is an urgent need for generic OoC platforms based on interoperability and modularity. Technology platforms based on open designs would enable seamless integration of diverse OoC models and components, facilitating translation. Our study introduces a modular microfluidic platform that integrates swappable modules for pumping, sensing, and OoCs, all within the ANSI/SLAS microplate footprint. Sub-components operate as microfluidic building blocks (MFBBs) and can interface with the demonstrated fluidic circuit board (FCB) universally as long as the designs adhere to ISO standards. The platform architecture allows tube-less inter-module interactions via arbitrary and reconfigurable fluidic circuits. We demonstrate two possible fluidic configurations which include in-line sensors and furthermore demonstrate biological functionality by running both in vitro and ex vivo OoC models for multiple days. This platform is designed to support automated multi-organ experiments, independent of the OoC type or material. All designs shown are made open source to encourage broader compatibility and collaboration.
{"title":"STARTER: a stand-alone reconfigurable and translational organ-on-chip platform based on modularity and open design principles","authors":"Aniruddha Paul, Eric R. Safai, Laura E. de Heus, Anke R. Vollertsen, Kevin Weijgertse, Bjorn de Wagenaar, Hossein E. Amirabadi, Evita van de Steeg, Mathieu Odijk, Andries D. van der Meer and Joshua Loessberg-Zahl","doi":"10.1039/D5LC00756A","DOIUrl":"10.1039/D5LC00756A","url":null,"abstract":"<p >Organ-on-chips (OoC) have the potential to revolutionize drug testing. However, the fragmented landscape of existing OoC systems leads to wasted resources and collaboration barriers, slowing broader adoption. To unite the ecosystem, there is an urgent need for generic OoC platforms based on interoperability and modularity. Technology platforms based on open designs would enable seamless integration of diverse OoC models and components, facilitating translation. Our study introduces a modular microfluidic platform that integrates swappable modules for pumping, sensing, and OoCs, all within the ANSI/SLAS microplate footprint. Sub-components operate as microfluidic building blocks (MFBBs) and can interface with the demonstrated fluidic circuit board (FCB) universally as long as the designs adhere to ISO standards. The platform architecture allows tube-less inter-module interactions <em>via</em> arbitrary and reconfigurable fluidic circuits. We demonstrate two possible fluidic configurations which include in-line sensors and furthermore demonstrate biological functionality by running both <em>in vitro</em> and <em>ex vivo</em> OoC models for multiple days. This platform is designed to support automated multi-organ experiments, independent of the OoC type or material. All designs shown are made open source to encourage broader compatibility and collaboration.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 3","pages":" 604-617"},"PeriodicalIF":5.4,"publicationDate":"2026-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2026/lc/d5lc00756a?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146044774","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}
The development of effective cancer therapies remains constrained by the complex and dynamic nature of the tumor microenvironment (TME), with tumor vasculature representing a critical barrier and modulator of treatment response. This review critically examines recent advances in the generation of vascularized tumor models using organ-on-a-chip (OoC) microfluidic technologies, emphasizing their capacity to recapitulate key interactions between tumor cells, stroma, and vasculature in vitro. We outline the mechanistic roles of tumor vasculature in therapy resistance, metastatic dissemination, and immune modulation, and highlight current strategies targeting vasculature for improved therapeutic outcomes. State-of-the-art biomaterials and engineering approaches, including template-based fabrication, self-organization, and the integration of patient-derived organoids, are discussed regarding their efficacy in constructing physiologically relevant vasculature. The review critically assesses findings from drug testing studies and discusses the translational potential of microfluidic platform capabilities, such as real-time monitoring, precise flow control, and functional assessment of vessel permeability and drug delivery, while identifying key limitations for clinical implementation. Challenges in standardization, scalability, and clinical translation are discussed, and recommendations are proposed to enhance the human-relevance and impact of vascularized OoC models in preclinical oncology research. These advanced platforms represent a transformative approach for bridging the translational gap between preclinical research and clinical oncology, offering opportunities to advance personalized cancer therapeutics and improve patient outcomes.
{"title":"Engineering perfusion to meet tumor biology: are vascularized tumor-on-a-chip models ready to drive therapy innovation?","authors":"Ines Poljak, Ciro Chiappini, Giulia Adriani","doi":"10.1039/d5lc01060h","DOIUrl":"https://doi.org/10.1039/d5lc01060h","url":null,"abstract":"The development of effective cancer therapies remains constrained by the complex and dynamic nature of the tumor microenvironment (TME), with tumor vasculature representing a critical barrier and modulator of treatment response. This review critically examines recent advances in the generation of vascularized tumor models using organ-on-a-chip (OoC) microfluidic technologies, emphasizing their capacity to recapitulate key interactions between tumor cells, stroma, and vasculature <em>in vitro</em>. We outline the mechanistic roles of tumor vasculature in therapy resistance, metastatic dissemination, and immune modulation, and highlight current strategies targeting vasculature for improved therapeutic outcomes. State-of-the-art biomaterials and engineering approaches, including template-based fabrication, self-organization, and the integration of patient-derived organoids, are discussed regarding their efficacy in constructing physiologically relevant vasculature. The review critically assesses findings from drug testing studies and discusses the translational potential of microfluidic platform capabilities, such as real-time monitoring, precise flow control, and functional assessment of vessel permeability and drug delivery, while identifying key limitations for clinical implementation. Challenges in standardization, scalability, and clinical translation are discussed, and recommendations are proposed to enhance the human-relevance and impact of vascularized OoC models in preclinical oncology research. These advanced platforms represent a transformative approach for bridging the translational gap between preclinical research and clinical oncology, offering opportunities to advance personalized cancer therapeutics and improve patient outcomes.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"51 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146044737","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}
Peilong Li, Yunfan Li, Jiajie Zhan, Deng Wang, Ruyu Zhang, Feng Liu
Microfluidic lab-on-a-chip technology has shown great potential in various fields such as bioscience, medical diagnostics, and environmental monitoring. However, its widespread adoption has been hindered by challenges in functional integration, operational autonomy, and manufacturing scalability. To address these limitations, we present a 3D-printed self-sensing magnetically actuated microfluidic (SMAM) chip designed for autonomous bioanalysis. This innovative device utilizes stereolithography apparatus (SLA) 3D printing to rapidly prototype and integrate microchannel networks alongside with a magnetically driven functional module. The chip employs magnetic actuation for precise, wireless manipulation of fluids within the microchannels, eliminating the need for bulky external pumps. Additionally, the system features an integrated self-sensing mechanism, enabling flow monitoring and on-chip analyte detection. The SMAM chip demonstrates exceptional dual-function performance, achieving a high pumping flow rate of up to 972 µL/min and a good piezoresistive sensitivity of 43.1 MPa⁻¹. We first demonstrate its system-level utility by assembling the chip into a modular, wirelessly monitored microfluidic platform with an integrated flow rectifier. Furthermore, its potential for therapeutic interventions is validated through a proof-of-concept of an untethered device for magnetically guided, on-demand drug release. This work provides a novel approach for developing intelligent analytical devices, promising to enable new paradigms in automated biological research and diagnostics.
{"title":"3D-printed self-sensing magnetically actuated microfluidic chip for closed-loop drug delivery","authors":"Peilong Li, Yunfan Li, Jiajie Zhan, Deng Wang, Ruyu Zhang, Feng Liu","doi":"10.1039/d5lc01006c","DOIUrl":"https://doi.org/10.1039/d5lc01006c","url":null,"abstract":"Microfluidic lab-on-a-chip technology has shown great potential in various fields such as bioscience, medical diagnostics, and environmental monitoring. However, its widespread adoption has been hindered by challenges in functional integration, operational autonomy, and manufacturing scalability. To address these limitations, we present a 3D-printed self-sensing magnetically actuated microfluidic (SMAM) chip designed for autonomous bioanalysis. This innovative device utilizes stereolithography apparatus (SLA) 3D printing to rapidly prototype and integrate microchannel networks alongside with a magnetically driven functional module. The chip employs magnetic actuation for precise, wireless manipulation of fluids within the microchannels, eliminating the need for bulky external pumps. Additionally, the system features an integrated self-sensing mechanism, enabling flow monitoring and on-chip analyte detection. The SMAM chip demonstrates exceptional dual-function performance, achieving a high pumping flow rate of up to 972 µL/min and a good piezoresistive sensitivity of 43.1 MPa⁻¹. We first demonstrate its system-level utility by assembling the chip into a modular, wirelessly monitored microfluidic platform with an integrated flow rectifier. Furthermore, its potential for therapeutic interventions is validated through a proof-of-concept of an untethered device for magnetically guided, on-demand drug release. This work provides a novel approach for developing intelligent analytical devices, promising to enable new paradigms in automated biological research and diagnostics.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"4 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146034221","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}
Guohao Yang, Seonghun Shin, Seongsu Cho, Jinkee Lee, Ryungeun Song
Three-dimensional (3D) printing has emerged as a promising method for fabricating microfluidic devices due to its rapid prototyping, adaptability, and cost-effectiveness. However, the intrinsic hydrophobicity of commercial photocurable resins limits their ability to generate stable oil-in-water (O/W) emulsions droplets. In this study, we addressed this limitation by introducing a simple yet effective surface modification technique, photochemical grafting, which covalently attaches hydrophilic methacrylic acid group onto the surfaces of 3D-printed channels, enabling reliable monodisperse O/W droplets formation. Integrating two modules with contrasting wettabilities yields a modular platform for single-step production of double emulsions (W/O/W and O/W/O). The result is a versatile system with precise control over droplet formation and exceptional monodispersity with tunable shell-to-core ratios. The grafted surfaces retained wettability and dropletgeneration performance after three months of storage and 15 hours of continuous shear. Collectively, this work presents a robust and scalable strategy to bridge rapid 3D printing with durable surface functionalization, expanding the potential of customizable emulsion generation in lab-on-a-chip applications.
{"title":"Surface modification of 3D printed microfluidic device by photochemical grafting","authors":"Guohao Yang, Seonghun Shin, Seongsu Cho, Jinkee Lee, Ryungeun Song","doi":"10.1039/d5lc00994d","DOIUrl":"https://doi.org/10.1039/d5lc00994d","url":null,"abstract":"Three-dimensional (3D) printing has emerged as a promising method for fabricating microfluidic devices due to its rapid prototyping, adaptability, and cost-effectiveness. However, the intrinsic hydrophobicity of commercial photocurable resins limits their ability to generate stable oil-in-water (O/W) emulsions droplets. In this study, we addressed this limitation by introducing a simple yet effective surface modification technique, photochemical grafting, which covalently attaches hydrophilic methacrylic acid group onto the surfaces of 3D-printed channels, enabling reliable monodisperse O/W droplets formation. Integrating two modules with contrasting wettabilities yields a modular platform for single-step production of double emulsions (W/O/W and O/W/O). The result is a versatile system with precise control over droplet formation and exceptional monodispersity with tunable shell-to-core ratios. The grafted surfaces retained wettability and dropletgeneration performance after three months of storage and 15 hours of continuous shear. Collectively, this work presents a robust and scalable strategy to bridge rapid 3D printing with durable surface functionalization, expanding the potential of customizable emulsion generation in lab-on-a-chip applications.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"288 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146044775","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}
Single-cell protein profiling furnishes exclusive insights into understanding and describing phenotypic heterogeneity in large populations, garnering significant attention from researchers. However, investigating protein information at single-cell resolution has presented significant challenges on account of the small size of cells, low abundance of proteins and scarcity of sensitive analytical methods. Microfluidics has emerged as a powerful platform for single-cell protein profiling because it enables efficient single-cell isolation, high-throughput processing of small-volume samples, and integrated microscale reactions in a user-friendly format. This review provides a broad perspective on the leading-edge microfluidic platforms for single-cell protein profiling. It showcases different microfluidic layouts for single-cell separation and explores how cutting-edge analysis techniques are integrated with these platforms for protein profiling. Furthermore, the potential challenges and future trends of microfluidics-based single-cell protein profiling are presented and evaluated.
{"title":"Single-cell protein profiling energized by microfluidic technology.","authors":"Ruizhe Yang,Qingyu Ruan,Wenshang Guo,Haicong Shen,Xiaoye Lin,Yingwen Chen,Ye Tao,Chaoyong Yang,Yukun Ren","doi":"10.1039/d5lc00854a","DOIUrl":"https://doi.org/10.1039/d5lc00854a","url":null,"abstract":"Single-cell protein profiling furnishes exclusive insights into understanding and describing phenotypic heterogeneity in large populations, garnering significant attention from researchers. However, investigating protein information at single-cell resolution has presented significant challenges on account of the small size of cells, low abundance of proteins and scarcity of sensitive analytical methods. Microfluidics has emerged as a powerful platform for single-cell protein profiling because it enables efficient single-cell isolation, high-throughput processing of small-volume samples, and integrated microscale reactions in a user-friendly format. This review provides a broad perspective on the leading-edge microfluidic platforms for single-cell protein profiling. It showcases different microfluidic layouts for single-cell separation and explores how cutting-edge analysis techniques are integrated with these platforms for protein profiling. Furthermore, the potential challenges and future trends of microfluidics-based single-cell protein profiling are presented and evaluated.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"35 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146021594","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}
Jueming Chen,Xiaogang Wang,Weijie Ye,Hui Kang,Siyan Xiao,Jiayu Li,Lihui Wang,Dongguo Lin,Dayu Liu
Tumor stem cells (TSCs) play a pivotal role in the development of tumor organoids. Consequently, the development of effective methods for the isolation and differential induction of TSCs is essential for the establishment of tumor organoids. In this study, we demonstrate a microfluidic single-cell culture technique that facilitates the selective expansion of TSCs and the subsequent generation of tumor organoids. Our findings demonstrate that microfluidic single-cell culture enables the generation of single-cell-derived tumorspheres (SDTs) across a variety of tumor cell lines of various tissue origins. The SDT cells exhibited definitive stem cell characteristics, as confirmed by the expression of stemness markers and functional cellular assays. Furthermore, the differential induction of individual TSCs resulted in the formation of single-cell-derived tumor organoids (STOs). The suitability of a microfluidic single-cell culture approach for patient-derived tumor specimens was also evaluated. Specifically, TSCs were successfully expanded from 16/26 primary colorectal cancer specimens, with SDT formation rates ranging from 0.02% to 17.77%. Differential induction culture of individual TSCs yielded enhanced STO formation efficiencies (25.02% to 65.30%). Collectively, these results establish microfluidic single-cell culture as a robust and adaptable methodology for TSC expansion and tumor organoid generation, offering a valuable platform to advance the field of tumor organoid engineering.
{"title":"Microfluidic single-cell culture represents a versatile approach for tumor stem cell expansion and tumor organoid generation.","authors":"Jueming Chen,Xiaogang Wang,Weijie Ye,Hui Kang,Siyan Xiao,Jiayu Li,Lihui Wang,Dongguo Lin,Dayu Liu","doi":"10.1039/d5lc00996k","DOIUrl":"https://doi.org/10.1039/d5lc00996k","url":null,"abstract":"Tumor stem cells (TSCs) play a pivotal role in the development of tumor organoids. Consequently, the development of effective methods for the isolation and differential induction of TSCs is essential for the establishment of tumor organoids. In this study, we demonstrate a microfluidic single-cell culture technique that facilitates the selective expansion of TSCs and the subsequent generation of tumor organoids. Our findings demonstrate that microfluidic single-cell culture enables the generation of single-cell-derived tumorspheres (SDTs) across a variety of tumor cell lines of various tissue origins. The SDT cells exhibited definitive stem cell characteristics, as confirmed by the expression of stemness markers and functional cellular assays. Furthermore, the differential induction of individual TSCs resulted in the formation of single-cell-derived tumor organoids (STOs). The suitability of a microfluidic single-cell culture approach for patient-derived tumor specimens was also evaluated. Specifically, TSCs were successfully expanded from 16/26 primary colorectal cancer specimens, with SDT formation rates ranging from 0.02% to 17.77%. Differential induction culture of individual TSCs yielded enhanced STO formation efficiencies (25.02% to 65.30%). Collectively, these results establish microfluidic single-cell culture as a robust and adaptable methodology for TSC expansion and tumor organoid generation, offering a valuable platform to advance the field of tumor organoid engineering.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"47 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146021595","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 kidney organ-on-a-chip (OoC) is a powerful tool for studying drug-induced nephrotoxicity, but its application is limited by the absence of liver metabolism and low throughput. Here, we developed a high-throughput liver-kidney OoC system (HLKOC) featuring microfluidics, plug-in biomimetic cups (MIMICups), scalable flow channel plates, precision-cut liver slices (PCLS), and 3D HK-2 cell spheroids. We first established a functional endothelial barrier by optimizing cell types, biomimetic blood flow rate, serum content, and membrane pore size. The structural and functional integrity of the PCLS and HK-2 spheroids within the MIMICups was then confirmed through histological staining, metabolic assays, and functional tests for viability, polarization, and transport. To evaluate the system’s utility, we integrated the HLKOC with a single kidney OoC control and multidisciplinary techniques—including biochemical analysis, computational toxicology, molecular docking, metabolomics, and transcriptomics—to investigate the nephrotoxicity of triptolide (TPL) and its underlying mechanisms. Results showed that, compared to the single kidney OoC, the HLKOC exhibited higher levels of urea, total protein, and albumin in the biomimetic blood, confirming the robust biosynthetic capacity of the PCLS-based liver chip and its ability to better simulate in vivo conditions. Notably, a TPL-induced elevation in urea was observed only in the HLKOC, demonstrating the superior sensitivity of the liver-kidney co-culture. Multi-omics analysis revealed that TPL induced distinct metabolic and transcriptional responses in the HLKOC, involving pathways related to linoleic acid metabolism and vesicle-mediated processes, and led to the significant downregulation of transport proteins Cubilin and GLUT1. These findings highlight the advantages of the HLKOC over single-organ systems for drug toxicity assessment and provide new insights into the mechanisms of TPL-induced nephrotoxicity.
{"title":"A High-throughput Liver-Kidney Metabolic Interaction Chip for Insights into the Nephrotoxicity Mechanisms of Triptolide","authors":"Siyu Liu, Yun Yang, Yifei Yang, Guangfei Wei, Liu Zhou, Jiawei Lin, Zheng Yuan, Yingfei Li, Zhe Wu, Ting Liu, Guozhuang Zhang","doi":"10.1039/d5lc01051a","DOIUrl":"https://doi.org/10.1039/d5lc01051a","url":null,"abstract":"The kidney organ-on-a-chip (OoC) is a powerful tool for studying drug-induced nephrotoxicity, but its application is limited by the absence of liver metabolism and low throughput. Here, we developed a high-throughput liver-kidney OoC system (HLKOC) featuring microfluidics, plug-in biomimetic cups (MIMICups), scalable flow channel plates, precision-cut liver slices (PCLS), and 3D HK-2 cell spheroids. We first established a functional endothelial barrier by optimizing cell types, biomimetic blood flow rate, serum content, and membrane pore size. The structural and functional integrity of the PCLS and HK-2 spheroids within the MIMICups was then confirmed through histological staining, metabolic assays, and functional tests for viability, polarization, and transport. To evaluate the system’s utility, we integrated the HLKOC with a single kidney OoC control and multidisciplinary techniques—including biochemical analysis, computational toxicology, molecular docking, metabolomics, and transcriptomics—to investigate the nephrotoxicity of triptolide (TPL) and its underlying mechanisms. Results showed that, compared to the single kidney OoC, the HLKOC exhibited higher levels of urea, total protein, and albumin in the biomimetic blood, confirming the robust biosynthetic capacity of the PCLS-based liver chip and its ability to better simulate in vivo conditions. Notably, a TPL-induced elevation in urea was observed only in the HLKOC, demonstrating the superior sensitivity of the liver-kidney co-culture. Multi-omics analysis revealed that TPL induced distinct metabolic and transcriptional responses in the HLKOC, involving pathways related to linoleic acid metabolism and vesicle-mediated processes, and led to the significant downregulation of transport proteins Cubilin and GLUT1. These findings highlight the advantages of the HLKOC over single-organ systems for drug toxicity assessment and provide new insights into the mechanisms of TPL-induced nephrotoxicity.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"87 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146021914","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}
solomon Oshabaheebwa, Utku Goreke, Yuxuan Du, Christopher L. Wirth, Zoe Sekyonda, Bryan Benson, Payam Fadaei, Yusang B. Ley, Nathan M Perez, Petros Giannikopoulos, David Nguyen, Michael A. Suster, Pedram Mohseni, Umut A. Gurkan
Emerging therapies in sickle cell disease (SCD) aim to restore healthy red blood cell (RBC) function, but they often yield heterogeneous cellular responses. There are no proven techniques to evaluate restored rheological functionality and heterogeneity in these RBCs. We present a biomimetic microcapillary network, high-speed imaging, and computational algorithms to analyze RBC capillary velocity profiles of the entire sample population at single-cell resolution. Using peripheral RBCs from SCD patients and healthy donors, we showed that RBC capillary transit velocity correlated with cell shape, hydrodynamic adaptability, and elongation index. Healthy RBCs exhibited a velocity distribution skewed toward higher values, whereas RBCs from individuals with SCD showed a shift toward lower velocities. SCD samples had a greater fraction of slow RBCs than healthy controls (42.1% ± 12.0% vs. 19.0% ± 4.9%, p<0.0001). We tested mixtures of healthy and SCD RBCs to simulate heterogeneous therapeutic effects and demonstrated that the assay was sensitive to small fractions of abnormal RBCs. The slow RBC fraction emerged as a potential biomarker associated with SCD disease severity. This fraction significantly increased under hypoxia showing sensitivity to hypoxia-induced sickling. Finally, we assessed in vitro-derived RBCs and observed distinct velocity profiles for nucleated and enucleated cells. Processing methods to enrich enucleated RBCs improved the velocity profile, producing a distribution that was more comparable to that of peripheral RBCs. This platform’s ability to assess individual RBCs and generate a velocity profile from a small number of cells makes it ideal for evaluating the rheological properties of in vitro-derived RBCs.
镰状细胞病（SCD）的新疗法旨在恢复健康的红细胞（RBC）功能，但它们往往产生异质细胞反应。目前还没有成熟的技术来评估这些红细胞恢复的流变功能和异质性。我们提出了一种仿生微毛细管网络，高速成像和计算算法，以单细胞分辨率分析整个样品群体的RBC毛细管速度曲线。利用SCD患者和健康供者的外周血红细胞，我们发现红细胞毛细血管传输速度与细胞形状、流体动力学适应性和延伸指数相关。健康红细胞的流速分布倾向于较高的值，而SCD患者的红细胞流速则倾向于较低的值。SCD样本中慢红细胞的比例高于健康对照组（42.1%±12.0% vs. 19.0%±4.9%,p<0.0001）。我们测试了健康红细胞和SCD红细胞的混合物，以模拟不同的治疗效果，并证明该方法对一小部分异常红细胞敏感。缓慢RBC分数成为与SCD疾病严重程度相关的潜在生物标志物。在低氧条件下，这一比例显著增加，表明对低氧诱导的镰状细胞敏感。最后，我们评估了体外衍生的红细胞，并观察到有核细胞和去核细胞的不同速度分布。浓缩无核红细胞的处理方法改善了速度剖面，产生的分布与周围红细胞的分布更相似。该平台能够评估单个红细胞，并从少量细胞中生成速度曲线，这使得它非常适合评估体外衍生红细胞的流变特性。
{"title":"Microfluidic capillary transit velocity as a functional measure for sickle cell disease and in vitro-derived red blood cells","authors":"solomon Oshabaheebwa, Utku Goreke, Yuxuan Du, Christopher L. Wirth, Zoe Sekyonda, Bryan Benson, Payam Fadaei, Yusang B. Ley, Nathan M Perez, Petros Giannikopoulos, David Nguyen, Michael A. Suster, Pedram Mohseni, Umut A. Gurkan","doi":"10.1039/d5lc00769k","DOIUrl":"https://doi.org/10.1039/d5lc00769k","url":null,"abstract":"Emerging therapies in sickle cell disease (SCD) aim to restore healthy red blood cell (RBC) function, but they often yield heterogeneous cellular responses. There are no proven techniques to evaluate restored rheological functionality and heterogeneity in these RBCs. We present a biomimetic microcapillary network, high-speed imaging, and computational algorithms to analyze RBC capillary velocity profiles of the entire sample population at single-cell resolution. Using peripheral RBCs from SCD patients and healthy donors, we showed that RBC capillary transit velocity correlated with cell shape, hydrodynamic adaptability, and elongation index. Healthy RBCs exhibited a velocity distribution skewed toward higher values, whereas RBCs from individuals with SCD showed a shift toward lower velocities. SCD samples had a greater fraction of slow RBCs than healthy controls (42.1% ± 12.0% vs. 19.0% ± 4.9%, p<0.0001). We tested mixtures of healthy and SCD RBCs to simulate heterogeneous therapeutic effects and demonstrated that the assay was sensitive to small fractions of abnormal RBCs. The slow RBC fraction emerged as a potential biomarker associated with SCD disease severity. This fraction significantly increased under hypoxia showing sensitivity to hypoxia-induced sickling. Finally, we assessed in vitro-derived RBCs and observed distinct velocity profiles for nucleated and enucleated cells. Processing methods to enrich enucleated RBCs improved the velocity profile, producing a distribution that was more comparable to that of peripheral RBCs. This platform’s ability to assess individual RBCs and generate a velocity profile from a small number of cells makes it ideal for evaluating the rheological properties of in vitro-derived RBCs.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"31 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146021915","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 intrinsic elasticity of the cell membrane cortex complex, i.e., cell surface, is a promising biomarker for cell status and disease, and has widespread biological and biomedical applications. However, measuring cell surface elasticity in real-time with high throughput has not been achieved so far. Here we develop a system and demonstrate that it can characterise the intrinsic surface elasticity of up to 411 cells per second, with a low latency of less than 1 millisecond per cell from an image to predicted elasticity. Our key innovation is to integrate a multi-layer perception (MLP) based machine learning algorithm, which infers the surface elasticity of cells from their camera-recorded steady-deformation profiles in a microchannel, with a high-fidelity mechanistic model, which resolves the cell surface, cytoplasm and nucleus and can accurately predict the flow-induced cell deformation. Applied to human prostate cancer PC-3 and leukaemia K-562 cell lines, the system enables measuring tens of thousands of cells within minutes, to explore the cell mechano-heterogeneity, the relation between surface elasticity and cell size, and the possibility of using surface elasticity and cell size for cell classification. We show that the measured cell surface elasticity is little affected by flow condition, when doubling the flow speed or suspension fluid viscosity. The system is also sensitive enough to detect a reduction of cell surface elasticity as a result of the cytochalasin D-induced actin disassembly. By enabling real-time high-throughput characterisation of the surface elasticity of cells, the present method may inspire new applications.
{"title":"Real-time high-throughput characterisation of the surface elasticity of suspended cells","authors":"Ziyu Guo, Yi Sui, Wen Wang","doi":"10.1039/d5lc00909j","DOIUrl":"https://doi.org/10.1039/d5lc00909j","url":null,"abstract":"The intrinsic elasticity of the cell membrane cortex complex, i.e., cell surface, is a promising biomarker for cell status and disease, and has widespread biological and biomedical applications. However, measuring cell surface elasticity in real-time with high throughput has not been achieved so far. Here we develop a system and demonstrate that it can characterise the intrinsic surface elasticity of up to 411 cells per second, with a low latency of less than 1 millisecond per cell from an image to predicted elasticity. Our key innovation is to integrate a multi-layer perception (MLP) based machine learning algorithm, which infers the surface elasticity of cells from their camera-recorded steady-deformation profiles in a microchannel, with a high-fidelity mechanistic model, which resolves the cell surface, cytoplasm and nucleus and can accurately predict the flow-induced cell deformation. Applied to human prostate cancer PC-3 and leukaemia K-562 cell lines, the system enables measuring tens of thousands of cells within minutes, to explore the cell mechano-heterogeneity, the relation between surface elasticity and cell size, and the possibility of using surface elasticity and cell size for cell classification. We show that the measured cell surface elasticity is little affected by flow condition, when doubling the flow speed or suspension fluid viscosity. The system is also sensitive enough to detect a reduction of cell surface elasticity as a result of the cytochalasin D-induced actin disassembly. By enabling real-time high-throughput characterisation of the surface elasticity of cells, the present method may inspire new applications.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"9 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146021977","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}
Lucas Poncelet, Keith J Morton, Matthew Shiu, Gaétan Veilleux, Chantal Richer, Liviu Clime, Daniel Sinnett, Teodor Veres
Extracellular vesicles (EVs), especially the exosome sized subset are increasingly exploited as minimally invasive cancer biomarkers. These small vesicles are abundant in biofluids and play crucial roles in intercellular communication and disease progression by transporting bioactive molecules. Exosomes offer distinct diagnostic and prognostic advantages over traditional cancer biomarkers, but purifying and extracting exosomes from blood remains challenging. There is a need to simply and cost-effectively isolate exosomes from milliliter quantities of whole blood for transcriptional and other omics-based research. Addressing this gap, we propose a microfluidic cartridge, the EV-Blade, for size and affinity-based purification of exosomes on a centrifugal microfluidic platform. We demonstrate a method to automate exosome purification from whole blood samples on a single microfluidic cartridge. The EV-Blade system combines blood centrifugation, plasma filtration for EV size selection and immunomagnetic capture using functionalized superparamagnetic nanoparticles targeting CD9, CD63, and CD81 exosomal surface proteins. We report on the device performance, purity of exosome recovery and the quality of RNA collected following on-chip EV lysis. We use this automated method to detect relevant long coding and non-coding RNA transcripts in circulating blood exosomes, showcasing the EV-Blade for use in cancer patient risk stratification. The system presented herein represents a significant advancement in exosome purification, offering a robust and automated platform for liquid biopsy-based cancer research and clinical applications. This innovation holds promise for cancer diagnosis, prognosis, and monitoring through non-invasive biomarkers.
{"title":"EV-Blade: an automated centrifugal-pneumatic cartridge for size- and affinity-based exosome isolation from whole blood","authors":"Lucas Poncelet, Keith J Morton, Matthew Shiu, Gaétan Veilleux, Chantal Richer, Liviu Clime, Daniel Sinnett, Teodor Veres","doi":"10.1039/d5lc00977d","DOIUrl":"https://doi.org/10.1039/d5lc00977d","url":null,"abstract":"Extracellular vesicles (EVs), especially the exosome sized subset are increasingly exploited as minimally invasive cancer biomarkers. These small vesicles are abundant in biofluids and play crucial roles in intercellular communication and disease progression by transporting bioactive molecules. Exosomes offer distinct diagnostic and prognostic advantages over traditional cancer biomarkers, but purifying and extracting exosomes from blood remains challenging. There is a need to simply and cost-effectively isolate exosomes from milliliter quantities of whole blood for transcriptional and other omics-based research. Addressing this gap, we propose a microfluidic cartridge, the EV-Blade, for size and affinity-based purification of exosomes on a centrifugal microfluidic platform. We demonstrate a method to automate exosome purification from whole blood samples on a single microfluidic cartridge. The EV-Blade system combines blood centrifugation, plasma filtration for EV size selection and immunomagnetic capture using functionalized superparamagnetic nanoparticles targeting CD9, CD63, and CD81 exosomal surface proteins. We report on the device performance, purity of exosome recovery and the quality of RNA collected following on-chip EV lysis. We use this automated method to detect relevant long coding and non-coding RNA transcripts in circulating blood exosomes, showcasing the EV-Blade for use in cancer patient risk stratification. The system presented herein represents a significant advancement in exosome purification, offering a robust and automated platform for liquid biopsy-based cancer research and clinical applications. This innovation holds promise for cancer diagnosis, prognosis, and monitoring through non-invasive biomarkers.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"101 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2026-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146001583","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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