Lab on a Chip最新文献

{"title":"Rapid miRNA detection in skin interstitial fluid using a hydrogel microneedle patch integrated with DNA probes and graphene oxide†","authors":"Hanjia Zheng, Fatemeh Keyvani, Sadegh Sadeghzadeh, Dragos F. Mantaila, Fasih A. Rahman, Joe Quadrilatero and Mahla Poudineh","doi":"10.1039/D4LC00715H","DOIUrl":"10.1039/D4LC00715H","url":null,"abstract":"<p >MicroRNA (miRNA) is a type of short, non-coding nucleic acid molecule that plays essential roles in diagnosing and prognosing various types of cancer. MiRNA is abundantly present in skin interstitial fluid (ISF), providing real-time and localized physiological information. Hydrogel microneedle (HMN) patches enable miRNA collection in a fast, pain-free, minimally invasive, and user-friendly manner. In this study, we introduced a fluorescence-based HMN assay, namely the HMN-miR sensor, composed of methacrylated hyaluronic acid (MeHA) and a graphene oxide–probe DNA (GO.pDNA) conjugate for miR21 and miR210 detection. The HMN-miR sensor demonstrates excellent skin penetration efficiency, rapid ISF collection capability, and sufficient miRNA detection and sequence identification specificity. The HMN-miR sensor facilitates a new assay that, with further optimization, could be applied in future clinical settings. Its simple fabrication process and excellent biocompatibility give it significant potential for various clinical uses, such as personalized cancer treatment and monitoring the healing progress of burn wounds.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 21","pages":" 4989-4997"},"PeriodicalIF":6.1,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142152385","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":"Rapid identification of bacterial isolates using microfluidic adaptive channels and multiplexed fluorescence microscopy†","authors":"Stelios Chatzimichail, Piers Turner, Conor Feehily, Alison Farrar, Derrick Crook, Monique Andersson, Sarah Oakley, Lucinda Barrett, Hafez El Sayyed, Jingwen Kyropoulos, Christoffer Nellåker, Nicole Stoesser and Achillefs N. Kapanidis","doi":"10.1039/D4LC00325J","DOIUrl":"10.1039/D4LC00325J","url":null,"abstract":"<p >We demonstrate the rapid capture, enrichment, and identification of bacterial pathogens using Adaptive Channel Bacterial Capture (ACBC) devices. Using controlled tuning of device backpressure in polydimethylsiloxane (PDMS) devices, we enable the controlled formation of capture regions capable of trapping bacteria from low cell density samples with near 100% capture efficiency. The technical demands to prepare such devices are much lower compared to conventional methods for bacterial trapping and can be achieved with simple benchtop fabrication methods. We demonstrate the capture and identification of seven species of bacteria with bacterial concentrations lower than 1000 cells per mL, including common Gram-negative and Gram-positive pathogens such as <em>Escherichia coli</em> and <em>Staphylococcus aureus</em>. We further demonstrate that species identification of the trapped bacteria can be undertaken in the order of one-hour using multiplexed 16S rRNA-FISH with identification accuracies of 70–98% with unsupervised classification methods across 7 species of bacteria. Finally, by using the bacterial capture capabilities of the ACBC chip with an ultra-rapid antimicrobial susceptibility testing method employing fluorescence imaging and convolutional neural network (CNN) classification, we demonstrate that we can use the ACBC chip as an imaging flow cytometer that can predict the antibiotic susceptibility of <em>E. coli</em> cells after identification.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 20","pages":" 4843-4858"},"PeriodicalIF":6.1,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d4lc00325j?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142152384","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":"CRISPR for companion diagnostics in low-resource settings†","authors":"Xu Qian, Qiang Xu, Christopher J. Lyon and Tony Y. Hu","doi":"10.1039/D4LC00340C","DOIUrl":"10.1039/D4LC00340C","url":null,"abstract":"<p >New point-of-care tests (POCTs), which are especially useful in low-resource settings, are needed to expand screening capacity for diseases that cause significant mortality: tuberculosis, multiple cancers, and emerging infectious diseases. Recently, clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostic (CRISPR-Dx) assays have emerged as powerful and versatile alternatives to traditional nucleic acid tests, revealing a strong potential to meet this need for new POCTs. In this review, we discuss CRISPR-Dx assay techniques that have been or could be applied to develop POCTs, including techniques for sample processing, target amplification, multiplex assay design, and signal readout. This review also describes current and potential applications for POCTs in disease diagnosis and includes future opportunities and challenges for such tests. These tests need to advance beyond initial assay development efforts to broadly meet criteria for use in low-resource settings.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 20","pages":" 4717-4740"},"PeriodicalIF":6.1,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d4lc00340c?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142138281","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":"Development of in vitro model of exosome transport in microfluidic gut-brain axis-on-a-chip","authors":"Gwang Myeong Seo, Hongki Lee, Yeon Jae Kang, Donghyun Kim and Jong Hwan Sung","doi":"10.1039/D4LC00490F","DOIUrl":"10.1039/D4LC00490F","url":null,"abstract":"<p >The gut communicates with the brain in a variety of ways known as the gut–brain axis (GBA), which is known to affect neurophysiological functions as well as neuronal disorders. Exosomes capable of passing through the blood–brain-barrier (BBB) have received attention as a mediator of gut–brain signaling and drug delivery vehicles. In conventional well plate-based experiments, it is difficult to observe the exosome movement in real time. Here, we developed a microfluidic-based GBA chip for co-culturing gut epithelial cells and neuronal cells and simultaneously observing exosome transport. The GBA-chip is aimed to mimic the <em>in vivo</em> situation of convective flow in blood vessels and convective and diffusive transport in the tissue interstitium. Here, fluorescence-labeled exosome was produced by transfection of HEK-293T cells with CD63-GFP plasmid. We observed in real time the secretion of CD63-GFP-exosomes by the transfected HEK-293T cells in the chip, and transport of the exosomes to neuronal cells and analyzed the dynamics of GFP-exosome movement. Our model is expected to enhance understanding of the roles of exosome in GBA.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 19","pages":" 4581-4593"},"PeriodicalIF":6.1,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142124251","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":"Development of an in vitro platform for the analysis of contractile and calcium dynamics in single human myotubes†","authors":"Camila Vesga-Castro, Laura Mosqueira-Martín, Paul Ubiria-Urkola, Pablo Marco-Moreno, Klaudia González-Imaz, Jorge Rendon-Hinestroza, Ainara Vallejo-Illarramendi and Jacobo Paredes","doi":"10.1039/D3LC00442B","DOIUrl":"10.1039/D3LC00442B","url":null,"abstract":"<p > <em>In vitro</em> myotube cultures are widely used as models for studying muscle pathophysiology, but their limited maturation and heterogeneity pose significant challenges for functional analyses. While they remain the gold standard for studying muscle function <em>in vitro</em>, myotube cultures do not fully recapitulate the complexity and native features of muscle fibers, which may compromise their ability to predict <em>in vivo</em> outcomes. To promote maturation and decrease heterogeneity, we have incorporated engineered structures into myotube cultures, based on a PDMS thin layer with micrometer-sized grooves (μGrooves) placed over a glass substrate. Different sizes and shapes of μGrooves were tested for their ability to promote alignment and fusion of myoblasts and enhance their differentiation into myotubes. A 24 hour electrical field stimulation protocol (4 V, 6 ms, 0.1 Hz) was used to further promote myotube maturation, after which several myotube features were assessed, including myotube alignment, width, fusion index, contractile function, and calcium handling. Our results indicate superior calcium and contractile performance in μGrooved myotubes, particularly with the 100 μm-width 700 μm-long geometry (7 : 1). This platform generated homogeneous and isolated myotubes that reproduced native muscle features, such as excitation–contraction coupling and force-frequency responses. Overall, our 2D muscle platform enables robust high-content assays of calcium dynamics and contractile readouts with increased sensitivity and reproducibility compared to traditional myotube cultures, making it particularly suitable for screening therapeutic candidates for different muscle pathologies.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 20","pages":" 4741-4754"},"PeriodicalIF":6.1,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d3lc00442b?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142130961","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":"Microfluidic technologies for lipid vesicle generation","authors":"Yu Cheng, Callum D. Hay, Suchaya M. Mahuttanatan, James W. Hindley, Oscar Ces and Yuval Elani","doi":"10.1039/D4LC00380B","DOIUrl":"10.1039/D4LC00380B","url":null,"abstract":"<p >Encapsulating biological and non-biological materials in lipid vesicles presents significant potential in both industrial and academic settings. When smaller than 100 nm, lipid vesicles and lipid nanoparticles are ideal vehicles for drug delivery, facilitating the delivery of payloads, improving pharmacokinetics, and reducing the off-target effects of therapeutics. When larger than 1 μm, vesicles are useful as model membranes for biophysical studies, as synthetic cell chassis, as bio-inspired supramolecular devices, and as the basis of protocells to explore the origin of life. As applications of lipid vesicles gain prominence in the fields of nanomedicine, biotechnology, and synthetic biology, there is a demand for advanced technologies for their controlled construction, with microfluidic methods at the forefront of these developments. Compared to conventional bulk methods, emerging microfluidic methods offer advantages such as precise size control, increased production throughput, high encapsulation efficiency, user-defined membrane properties (<em>i.e.</em>, lipid composition, vesicular architecture, compartmentalisation, membrane asymmetry, <em>etc.</em>), and potential integration with lab-on-chip manipulation and analysis modules. We provide a review of microfluidic lipid vesicle generation technologies, focusing on recent advances and state-of-the-art techniques. Principal technologies are described, and key research milestones are highlighted. The advantages and limitations of each approach are evaluated, and challenges and opportunities for microfluidic engineering of lipid vesicles to underpin a new generation of therapeutics, vaccines, sensors, and bio-inspired technologies are presented.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 20","pages":" 4679-4716"},"PeriodicalIF":6.1,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d4lc00380b?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142138313","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":"Investigating the impact of the interstitial fluid flow and hypoxia interface on cancer transcriptomes using a spheroid-on-chip perfusion system†","authors":"Emily Pyne, Mark Reardon, Martin Christensen, Pablo Rodriguez Mateos, Scott Taylor, Alexander Iles, Ananya Choudhury, Nicole Pamme and Isabel M. Pires","doi":"10.1039/D4LC00512K","DOIUrl":"10.1039/D4LC00512K","url":null,"abstract":"<p >Solid tumours are complex and heterogeneous systems, which exist in a dynamic biophysical microenvironment. Conventional cancer research methods have long relied on two-dimensional (2D) static cultures which neglect the dynamic, three-dimensional (3D) nature of the biophysical tumour microenvironment (TME), especially the role and impact of interstitial fluid flow (IFF). To address this, we undertook a transcriptome-wide analysis of the impact of IFF-like perfusion flow using a spheroid-on-chip microfluidic platform, which allows 3D cancer spheroids to be integrated into extracellular matrices (ECM)-like hydrogels and exposed to continuous perfusion, to mimic IFF in the TME. Importantly, we have performed these studies both in experimental (normoxia) and pathophysiological (hypoxia) oxygen conditions. Our data indicated that gene expression was altered by flow when compared to static conditions, and for the first time showed that these gene expression patterns differed in different oxygen tensions, reflecting a differential role of spheroid perfusion in IFF-like flow in tumour-relevant hypoxic conditions in the biophysical TME. We were also able to identify factors primarily linked with IFF-like conditions which are linked with prognostic value in cancer patients and therefore could correspond to a potential novel biomarker of IFF in cancer. This study therefore highlights the need to consider relevant oxygen conditions when studying the impact of flow in cancer biology, as well as demonstrating the potential of microfluidic models of flow to identify IFF-relevant tumour biomarkers.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 19","pages":" 4609-4622"},"PeriodicalIF":6.1,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d4lc00512k?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142130940","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":"Engineered cortical microcircuits for investigations of neuroplasticity†","authors":"Nicolai Winter-Hjelm, Pawel Sikorski, Axel Sandvig and Ioanna Sandvig","doi":"10.1039/D4LC00546E","DOIUrl":"10.1039/D4LC00546E","url":null,"abstract":"<p >Recent advances in neural engineering have opened new ways to investigate the impact of topology on neural network function. Leveraging microfluidic technologies, it is possible to establish modular circuit motifs that promote both segregation and integration of information processing in the engineered neural networks, similar to those observed <em>in vivo</em>. However, the impact of the underlying topologies on network dynamics and response to pathological perturbation remains largely unresolved. In this work, we demonstrate the utilization of microfluidic platforms with 12 interconnected nodes to structure modular, cortical engineered neural networks. By implementing geometrical constraints inspired by a Tesla valve within the connecting microtunnels, we additionally exert control over the direction of axonal outgrowth between the nodes. Interfacing these platforms with nanoporous microelectrode arrays reveals that the resulting laminar cortical networks exhibit pronounced segregated and integrated functional dynamics across layers, mirroring key elements of the feedforward, hierarchical information processing observed in the neocortex. The multi-nodal configuration also facilitates selective perturbation of individual nodes within the networks. To illustrate this, we induced hypoxia, a key factor in the pathogenesis of various neurological disorders, in well-connected nodes within the networks. Our findings demonstrate that such perturbations induce ablation of information flow across the hypoxic node, while enabling the study of plasticity and information processing adaptations in neighboring nodes and neural communication pathways. In summary, our presented model system recapitulates fundamental attributes of the microcircuit organization of neocortical neural networks, rendering it highly pertinent for preclinical neuroscience research. This model system holds promise for yielding new insights into the development, topological organization, and neuroplasticity mechanisms of the neocortex across the micro- and mesoscale level, in both healthy and pathological conditions.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 21","pages":" 4974-4988"},"PeriodicalIF":6.1,"publicationDate":"2024-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142130908","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":"From ex ovo to in vitro: xenotransplantation and vascularization of mouse embryonic kidneys in a microfluidic chip†","authors":"Micaela Oliveira, Partha Protim Sarker, Ilya Skovorodkin, Ali Kalantarifard, Tugce Haskavuk, Jonatan Mac Intyre, Elizabath Nallukunnel Raju, Samin Nooranian, Hiroki Shioda, Masaki Nishikawa, Yasuyuki Sakai, Seppo J. Vainio, Caglar Elbuken and Irina Raykhel","doi":"10.1039/D4LC00547C","DOIUrl":"10.1039/D4LC00547C","url":null,"abstract":"<p >Organoids are emerging as a powerful tool to investigate complex biological structures <em>in vitro</em>. Vascularization of organoids is crucial to recapitulate the morphology and function of the represented human organ, especially in the case of the kidney, whose primary function of blood filtration is closely associated with blood circulation. Current <em>in vitro</em> microfluidic approaches have only provided initial vascularization of kidney organoids, whereas <em>in vivo</em> transplantation to animal models is problematic due to ethical problems, with the exception of xenotransplantation onto a chicken chorioallantoic membrane (CAM). Although CAM can serve as a good environment for vascularization, it can only be used for a fixed length of time, limited by development of the embryo. Here, we propose a novel lab on a chip design that allows organoids of different origin to be cultured and vascularized on a CAM, as well as to be transferred to <em>in vitro</em> conditions when required. Mouse embryonic kidneys cultured on the CAM showed enhanced vascularization by intrinsic endothelial cells, and made connections with the chicken vasculature, as evidenced by blood flowing through them. After the chips were transferred to <em>in vitro</em> conditions, the vasculature inside the organoids was successfully maintained. To our knowledge, this is the first demonstration of the combination of <em>in vivo</em> and <em>in vitro</em> approaches applied to microfluidic chip design.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 20","pages":" 4816-4826"},"PeriodicalIF":6.1,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d4lc00547c?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142117987","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":"Micromixer driven by bubble-induced acoustic microstreaming for multi-ink 3D bioprinting†","authors":"Mitsuyuki Hidaka, Masaru Kojima and Shinji Sakai","doi":"10.1039/D4LC00552J","DOIUrl":"10.1039/D4LC00552J","url":null,"abstract":"<p >Recently, the 3D printing of cell-laden hydrogel structures, known as bioprinting, has received increasing attention owing to advances in tissue engineering and drug screening. However, a micromixing technology that efficiently mixes viscous bioinks under mild conditions is needed. Therefore, this study presents a novel method for achieving homogeneous mixing of multiple inks in 3D bioprinting through acoustic stimulation. This technique involves generating an acoustic microstream through bubble oscillations inside a 3D bioprinting nozzle. We determined the optimal hole design for trapping a bubble, hole arrangement, and voltage for efficient mixing, resulting in a four-fold increase in mixing efficiency compared to a single bubble arrangement. Subsequently, we propose a nozzle design for efficient mixing during bioprinting. The proposed nozzle design enabled the successful printing of line structures with a uniform mixture of different viscous bioinks, achieving a mixing efficiency of over 80% for mixing 0.5–1.0 wt% sodium alginate aqueous solutions. Additionally, acoustic stimulation had no adverse effects on cell viability, maintaining a high cell viability of 88% after extrusion. This study presents the first use of a bubble micromixer in 3D bioprinting, demonstrating gentle yet effective multi-ink mixing. We believe this approach will broaden 3D printing applications, particularly for constructing functional structures in 3D bioprinting.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 19","pages":" 4571-4580"},"PeriodicalIF":6.1,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142102314","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}