Lab on a Chip最新文献

{"title":"Sheath-enhanced concentration and on-chip detection of bacteria from an extremely low-concentration level.","authors":"Xinye Chen, Ruonan Peng, Ruo-Qian Wang, Ke Du","doi":"10.1039/d4lc00698d","DOIUrl":"https://doi.org/10.1039/d4lc00698d","url":null,"abstract":"<p><p>Microfluidic-based sheath flow focusing methods have been widely used for efficiently isolating, concentrating, and detecting pathogenic bacteria for various biomedical applications due to their enhanced sensitivity and exceptional integration. However, such a microfluidic device usually needs complicated device fabrication and sample dilution, hampering the efficient and sensitive identification of target bacteria. In this study, we develop and fabricate a sheath-assisted and pneumatic-induced nano-sieve device for achieving the improved on-chip concentration and sensitive detection of <i>Staphylococcus aureus</i> (MRSA). The optimized nanochannel design with diverging configuration is beneficial to the regulation of the hydrodynamic flow while the sheath flow is focusing the sample to the confined region as expected. Per the experimental finding, a high flow ratio (sheath flow/sample flow) presents enhanced target concentration by comparing with a low flow ratio. With this setup, MRSA bacteria with an extremely low concentration of ∼100 CFU mL^-1 are successfully and sensitively detected under a fluorescence microscope, less than 30 min, demonstrating a reliable sheath-enhanced concentration and on-chip detection for target bacteria. Additionally, the theoretical model introduced here further rationalizes the working principle of our nano-sieve device, potentially guiding the optimization of next generation devices for highly sensitive and accurate on-chip bacteria detection at a much lower concentration level below 100 CFU mL^-1.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142833109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An agarose fluidic chip for high-throughput in toto organoid imaging†","authors":"Sarah De Beuckeleer, Andres Vanhooydonck, Johanna Van Den Daele, Tim Van De Looverbosch, Bob Asselbergh, Hera Kim, Coen Campsteijn, Peter Ponsaerts, Regan Watts and Winnok H. De Vos","doi":"10.1039/D4LC00459K","DOIUrl":"10.1039/D4LC00459K","url":null,"abstract":"<p >Modern cell and developmental biology increasingly relies on 3D cell culture systems such as organoids. However, routine interrogation with microscopy is often hindered by tedious, non-standardized sample mounting, limiting throughput. To address these bottlenecks, we have developed a pipeline for imaging intact organoids in flow, utilizing a transparent agarose fluidic chip that enables efficient and consistent recordings with theoretically unlimited throughput. The chip, cast from a custom-designed 3D-printed mold, is coupled to a mechanically controlled syringe pump for fast and precise sample positioning. We benchmarked this setup on a commercial digitally scanned light sheet microscope with cleared glioblastoma spheroids. Spheroids of varying sizes were positioned in the field of view with micrometer-level stability, achieving a throughput of 40 one-minute recordings per hour. We further showed that sample positioning could be automated through online feedback microscopy. The optical quality of the agarose chip outperformed FEP tubing, glass channels and PDMS casts for the clearing agents used, as demonstrated by image contrast profiles of spheroids stained with a fluorescent nuclear counterstain and further emphasized by the resolution of fine microglial ramifications within cerebral organoids. The retention of image quality throughout 500 μm-sized spheroids enabled comprehensive spatial mapping of live and dead cells based on their nuclear morphology. Finally, imaging a batch of <em>LMNA</em> knockout <em>vs.</em> wildtype astrocytoma spheroids revealed significant differences in their DNA damage response, underscoring the system's sensitivity and throughput. Overall, the fluidic chip design provides a cost-effective, accessible, and efficient solution for high-throughput organoid imaging.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 2","pages":" 235-252"},"PeriodicalIF":6.1,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2025/lc/d4lc00459k?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142833100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Highly efficient combination of multiple single cells using a deterministic single-cell combinatorial reactor.","authors":"Mina Yoshida, Saori Tago, Kunihiko Iizuka, Teruo Fujii, Soo Hyeon Kim","doi":"10.1039/d4lc00951g","DOIUrl":"https://doi.org/10.1039/d4lc00951g","url":null,"abstract":"<p><p>Compartmentalization of multiple single cells and/or single microbeads holds significant potential for advanced biological research including single-cell transcriptome analysis or cell-cell interactions. To ensure reliable analysis and prevent misinterpretation, it is essential to achieve highly efficient pairing or combining of single objects. In this paper, we introduce a novel microfluidic device coupled with a multilayer interconnect Si/SiO² control circuit, named the deterministic single-cell combinatorial reactor (DSCR) device, for the highly efficient combination of multiple single cells. The deterministic combination of multiple single cells is realized by sequentially introducing and trapping each cell population into designated trap-wells within each DSCR. These cell-sized trap-wells, created by etching the SiO₂ passivation layer, generate a highly localized electric field that facilitates deterministic single-cell trapping. The device's multilayer interconnection of electrodes enables the sequential operation of each trap-well, allowing precise trapping of each cell population into designated trap-wells within an array of combinatorial reactors. We demonstrated the feasibility of the DSCR by sequentially trapping three distinct groups of PC3 cells, each stained with a different fluorescent dye (blue, green, or red). This method achieved a 93 ± 2% pairing efficiency for two cell populations and an 82 ± 7% combination efficiency for three cell populations. Our innovative system offers promising applications for analyzing multiple cell-cell communications and combinatorial indexing of single cells.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142826744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An intimal-lumen model in a microfluidic device: potential platform for atherosclerosis-related studies†","authors":"Fahima Akther, Dimple Sajin, Shehzahdi S. Moonshi, Jessica Pickett, Yuao Wu, Jun Zhang, Nam-Trung Nguyen and Hang Thu Ta","doi":"10.1039/D4LC00868E","DOIUrl":"10.1039/D4LC00868E","url":null,"abstract":"<p >Atherosclerosis is a chronic inflammatory vascular disorder driven by factors such as endothelial dysfunction, hypertension, hyperlipidemia, and arterial calcification, and is considered a leading global cause of death. Existing atherosclerosis models have limitations due to the absence of an appropriate hemodynamic microenvironment <em>in vitro</em> and interspecies differences <em>in vivo</em>. Here, we develop a simple but robust microfluidic intimal-lumen model of early atherosclerosis using interconnected dual channels for studying monocyte transmigration and foam cell formation at an arterial shear rate. To the best of our knowledge, this is the first study that creates a physiologically relevant microenvironment under an arterial shear rate to modulate lipid-laden foam cells on a microfluidic platform. As a proof of concept, we use murine endothelial cells to develop a vascular lumen in one channel and collagen-embedded murine smooth muscle cells to mimic the subendothelial intimal layer in another channel. The model successfully triggers endothelial dysfunction upon TNF-α stimulation, initiating monocyte adhesion to the endothelial monolayer under the arterial shear rate. Unlike existing <em>in vitro</em> models, native low-density lipoprotein (LDL) is added in the culture media instead of ox-LDL to stimulate subendothelial lipid accumulation, thereby mimicking more accurate physiology. The subendothelial transmigration of adherent monocytes and subsequent foam cell formation is also achieved under flow conditions in the model. The model also investigates the inhibitory effect of aspirin in monocyte adhesion and transmigration. The model exhibits a significant dose-dependent reduction in monocyte adhesion and transmigration upon aspirin treatment, making it an excellent tool for drug testing.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 3","pages":" 354-369"},"PeriodicalIF":6.1,"publicationDate":"2024-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142851733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A vascularized microfluidic model of the osteochondral unit for modeling inflammatory response and therapeutic screening†","authors":"Kevin D. Roehm, Irene Chiesa, Dustin Haithcock, Riccardo Gottardi and Balabhaskar Prabhakarpandian","doi":"10.1039/D4LC00651H","DOIUrl":"10.1039/D4LC00651H","url":null,"abstract":"<p >Osteoarthritis (OA) has long been considered a disease of the articular cartilage. Within the past decade it has become increasingly clear that OA is a disease of the entire joint space and that interactions between articular cartilage and subchondral bone likely play an important role in the disease. Driven by this knowledge, we have created a novel microphysiological model of the osteochondral unit containing synovium, cartilage, bone, and vasculature in separate compartments with molecular and direct cell–cell interaction between the cells from the different tissue types. We have characterized the model in terms of differentiation by molecule and matrix secretion and shown that it demonstrates morphology and functionality that mimic the native characteristic of the joint space. Finally, we induced inflammation and subsequently rescued the model constructs by a known compound as proof of concept for anti-inflammatory drug screening applications.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 3","pages":" 370-382"},"PeriodicalIF":6.1,"publicationDate":"2024-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142880747","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":"Beveled microneedles with channel for transdermal injection and sampling, fabricated with minimal steps and standard MEMS technology†","authors":"Alvise Bagolini, Nicolò G. Di Novo, Severino Pedrotti, Matteo Valt, Cristian Collini, Nicola M. Pugno and Leandro Lorenzelli","doi":"10.1039/D4LC00880D","DOIUrl":"10.1039/D4LC00880D","url":null,"abstract":"<p >Microneedles hold the potential for enabling shallow skin penetration applications where biomarkers are extracted from the interstitial fluid (ISF) and drugs are injected in a painless and effective manner. To this purpose, needles must have an inner channel. Channeled needles were demonstrated using custom silicon microtechnology, having several needle tip geometries. Nevertheless, all the proposed fabrication sequences are not compatible with mass production based on mature, standard microfabrication techniques. Furthermore, ISF extraction was also demonstrated with channeled needles but under poorly controlled conditions and over long periods of time, the latter being impractical for medical use. A range of factors may impede or slow ISF extraction that require controlled experiments. In this work we address the above tasks in terms of microfabrication sequence design, tip geometry design and experimental validation under controlled conditions. We report the development and fabrication of a silicon channeled microneedle array using conventional, industrial micromechanic processes. With only 2 lithography steps, a hypodermic needle tip profile is achieved. Using the fabricated microneedles, fluid extraction is experimented on chicken skin mockups. Extraction tests are carried out by inducing a controlled pressure gradient between the two ends of the microneedle channels, generated by loading the chip or by applying vacuum to the chip's backside. The extraction of more than 1 μL of fluid in 20 minutes is demonstrated with a maximum applied pressure gradient of 500 mbar. A correlation between the extraction rate efficiency and needles' density is observed, both for short and long extraction times. These results provide the first demonstration of <em>in vitro</em> interstitial fluid collection under controlled experimental conditions using silicon hollow microneedles fabricated with standard micro electro mechanical systems (MEMS) fabrication technology and minimal steps. Based on the obtained data, a comparison is drawn between pressure load and vacuum as drivers for ISF extraction, according to modelling and controlled experiments.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 2","pages":" 201-211"},"PeriodicalIF":6.1,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142811411","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":"Liquid metal electrodes enabled cascaded on-chip dielectrophoretic separation of large-size-range particles†","authors":"Huichao Chai, Liang Huang, Junwen Zhu, Jialu Tian and Wenhui Wang","doi":"10.1039/D4LC00942H","DOIUrl":"10.1039/D4LC00942H","url":null,"abstract":"<p >The separation of large-size-range particles of complex biological samples is critical but yet well resolved. As a label-free technique, dielectrophoresis (DEP)-based particle separation faces the challenge of how to configure DEP in an integrated microfluidic device to bring particles of various sizes into the effective DEP force field. Herein, we propose a concept that combines the passive flow fraction mechanism with the accumulative DEP deflection effect in a cascaded manner. This concept places DEP deflection segments and bypass outlets alternately. Each DEP deflection segment is configured with an array of side-wall liquid metal electrodes to exert effective DEP forces on the particles of a suitable size range. After each DEP deflection segment, the passive bypass flow fraction mechanism diverts part of the sample flow and target range of particles through the bypass outlet. Simultaneously, this flow fraction brings the remaining particles closer to the electrodes in the subsequent DEP deflection segment, causing the next size range of particles to deflect under effective DEP forces and thus making them separable. Repeating this process, particles would be separated from the bypass outlets one by one in the order of reducing size ranges. We present the concept design and modeling, and prove the concept through separating five different particles ranging from 16–0.5 μm mixed together to mimic blood composition, providing a powerful platform for separating multiple particles in diverse biomedical applications.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 3","pages":" 308-318"},"PeriodicalIF":6.1,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142875534","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":"Intelligent optoelectrowetting digital microfluidic system for real-time selective parallel manipulation of biological droplet arrays.","authors":"Tianyi Wang, Shizheng Zhou, Xuekai Liu, Jianghao Zeng, Xiaohan He, Zhihang Yu, Zhiyuan Liu, Xiaomei Liu, Jing Jin, Yonggang Zhu, Liuyong Shi, Hong Yan, Teng Zhou","doi":"10.1039/d4lc00804a","DOIUrl":"https://doi.org/10.1039/d4lc00804a","url":null,"abstract":"<p><p>Optoelectrowetting technology generates virtual electrodes to manipulate droplets by projecting optical patterns onto the photoconductive layer. This method avoids the complex design of the physical circuitry of dielectricwetting chips, compensating for the inability to reconstruct the electrode. However, the current technology relies on operators to manually position the droplets, draw optical patterns, and preset the droplet movement paths. It lacks real-time feedback on droplet information and the ability for independent droplet control, which can lead to droplet miscontrol and contamination. This paper presents a combination of optoelectrowetting with deep learning algorithms, integrating software and a photoelectric detection platform, and develops an optoelectrowetting intelligent control system. First, a target detection algorithm identifies droplet characteristics in real-time and automatically generate virtual electrodes to control movement. Simultaneously, a tracking algorithm outputs trajectories and ID information for efficient droplet arrays tracking. The results show that the system can automatically control the movement and fusion of multiple droplets in parallel and realize the automatic arrangement and storage of disordered droplet arrays without any additional electrodes and sensing devices. Additionally, through the automated control of the system, the cell suspension can be precisely cultured in the specified medium according to experimental requirements, and the growth trend is consistent with that observed in the well plate, significantly enhancing the experiment's flexibility and accuracy. In this paper, we propose an intelligent method applicable to the automated manipulation of discrete droplets. This method would play a crucial role in advancing the applications of digital microfluidic technology in biomedicine and other fields.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142805626","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":"Retinal organoid chip: engineering a physiomimetic oxygen gradient for optimizing long term culture of human retinal organoids.","authors":"Emma Drabbe, Daniel Pelaez, Ashutosh Agarwal","doi":"10.1039/d4lc00771a","DOIUrl":"10.1039/d4lc00771a","url":null,"abstract":"<p><p>An oxygen gradient across the retina plays a crucial role in its development and function. The inner retina resides in a hypoxic environment (2% O₂) adjacent to the vitreous cavity. Oxygenation levels rapidly increase towards the outer retina (18% O₂) at the choroid. In addition to retinal stratification, oxygen levels are critical for the health of retinal ganglion cells (RGCs), which relay visual information from the retina to the brain. Human stem cell derived retinal organoids are being engineered to mimic the structure and function of human retina for applications such as disease modeling, development of therapeutics, and cell replacement therapies. However, rapid degeneration of the retinal ganglion cell layers are a common limitation of human retinal organoid platforms. We report the design of a novel retinal organoid chip (ROC) that maintains a physiologically relevant oxygen gradient and allows the maturation of inner and outer retinal cell phenotypes for human retinal organoids. Our PDMS-free ROC holds 55 individual retinal organoids that were manually seeded, cultured for extended periods (over 150 days), imaged <i>in situ</i>, and retrieved. ROC was designed from first principles of liquid and gas mass transport, and fabricated from biologically- and chemically inert materials using rapid prototyping techniques such as micromachining, laser cutting, 3D printing and bonding. After computational and experimental validation of oxygen gradients, human induced pluripotent stem cell derived retinal organoids were transferred into the ROC, differentiated, cultured and imaged within the chip. ROCs that maintained active perfusion and stable oxygen gradients were successful in inducing higher viability of RGCs within retinal organoids than static controls, or ROC without oxygen gradients. Our physiologically relevant and higher-throughput retinal organoid culture system is well suited for applications in studying developmental perturbations to primate retinogenesis, including those driven by inherited traits, fetal environmental exposure to toxic agents, or acquired by genetic mutations, such as retinoblastoma.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11632457/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142805628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Injury-on-a-chip for modelling microvascular trauma-induced coagulation†","authors":"Halston Deal, Elizabeth M. Byrnes, Sanika Pandit, Anastasia Sheridan, Ashley C. Brown and Michael Daniele","doi":"10.1039/D4LC00471J","DOIUrl":"10.1039/D4LC00471J","url":null,"abstract":"<p >Blood coagulation is a highly regulated injury response that features polymerization of fibrin fibers to prevent the passage of blood from a damaged vascular endothelium. A growing body of research seeks to monitor coagulation in microfluidic systems but fails to capture coagulation as a response to disruption of the vascular endothelium. Here we present a device that allows compression injury of a defined segment of a microfluidic vascular endothelium and the assessment of coagulation at the injury site. This pressure injury-on-a-chip (PINCH) device allows visualization of coagulation as the accumulation of fluorescent fibrin at injury sites. Quantification of fluorescent fibrin levels upstream of and at injury sites confirm that pre-treating vascular endothelium with fluid shear stress helps capture coagulation as an injury response. We leverage the PINCH devices to demonstrate the limited coagulation response of type A hemophiliacs and evaluate the performance of hemostatic microparticles and fibrinolytic nanoparticles. Our findings and the straightforward fabrication of the PINCH devices make it a promising choice for additional screening of hemostatic therapeutics.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 3","pages":" 440-453"},"PeriodicalIF":6.1,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11704661/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142941512","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}