{"title":"Programmable Cell Culture Chips for Topographical Manipulation of Living Cells","authors":"Xin-Yi Wu, Jian-Miao Zhang, Meng-Yao Niu, Fan-Chun Bin, Qi Duan, Jie Liu, Xian-Zi Dong, Mei-Ling Zheng","doi":"10.1039/d5lc00803d","DOIUrl":"https://doi.org/10.1039/d5lc00803d","url":null,"abstract":"The micro-morphological characteristics of biomaterial surfaces play a critical role in influencing cell proliferation, adhesion, and differentiation. However, the underlying mechanisms by which surface features modulate cellular behavior remain inadequately understood. Moreover, current surface designs intended for cell regulation tend to be overly simplistic, often failing to meet the dual requirements of high-precision fabrication and structural versatility. Here, we propose a programmable cell culture chip based on femtosecond laser maskless optical projection lithography (Fs-MOPL) technology to modulate the cell behavior. The as-fabricated chip exhibits high structural fidelity and uniformity. Surface treatment with O2 plasma followed by poly-D-lysine (PDL) coating enhances hydrophilicity, cell adhesion and growth. We have investigated the migration, adhesion, and morphological changes of 786-O cells on scaffold with varied line spacing, column diameter and hole size using immunofluorescence staining and confocal fluorescence microscopy. The cells cultured on linear array structures display elongated, oriented actin stress fibers, while column and hole array structures influence focal adhesion distribution and cellular tension. Biocompatibility characterization further confirms the chip's suitability for cell culture applications. Our findings highlight the potential of programmable cell culture chips to mimic complex in vivo microenvironments, offering a multifunctional platform for studying cell behavior and advancing biomedical research.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"23 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145189168","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lab on a ChipPub Date : 2025-09-30DOI: 10.1039/d5lc00337g
Dong Hyun Han, Yurim Kim, Yu-Jin Park, Dong-Yeon Song, Jongho Park, Jeonghwan Oh, Dong-Myung Kim, Je-Kyun Park
{"title":"Pushbutton-activated microfluidic cell-free biosensor for multiplexed pathogen detection","authors":"Dong Hyun Han, Yurim Kim, Yu-Jin Park, Dong-Yeon Song, Jongho Park, Jeonghwan Oh, Dong-Myung Kim, Je-Kyun Park","doi":"10.1039/d5lc00337g","DOIUrl":"https://doi.org/10.1039/d5lc00337g","url":null,"abstract":"In this paper, we have developed a novel cell-free biosensor based on a multiplexed pushbutton-activated microfluidic device (mPAMD) that enables simultaneous detection of multiple 16S rRNAs of pathogens in a single device. The multi-step target-responsive cell-free protein synthesis process was seamlessly integrated into a single microfluidic device with an intuitive finger-pumping mechanism, allowing simultaneous mixing, aliquoting, and detection of 16S rRNAs through the production of reporter proteins. The mPAMD incorporates multiplexed detection zones with pathogen-specific probes to enable the identification of multiple 16S rRNAs that allow a simple and intuitive diagnostic platform for cell-free biosensors. Microchannels were designed and optimized to achieve efficient sample mixing and even distribution of common reagents, ensuring uniform reaction conditions across all reaction channels. The developed system achieved a detection limit for 16S rRNA ranging from 1.69 to 7.39 pM, corresponding to approximately 10<small><sup>4</sup></small> to 10<small><sup>5</sup></small> CFU/mL of pathogens. These results address the growing demand for an accessible multiplexed diagnostic system while ensuring high sensitivity and specificity.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"201 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145189442","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":"Electrical Impedance Tomography (EIT)-Based Intracellular Conductivity Imaging for Non-invasive Cell Detection","authors":"Songshi Li, Daisuke Kawashima, Zeyang Dai, Nobuyuki Aoki, Masahiro Takei","doi":"10.1039/d5lc00466g","DOIUrl":"https://doi.org/10.1039/d5lc00466g","url":null,"abstract":"Electrical impedance tomography (EIT)-based intracellular conductivity imaging is newly proposed as a non-invasive technique for mapping the electrical properties of living cells at the single-cell scale. In order to achieve this, a micro-EIT system is developed, which integrates two main components: a custom-designed micro-EIT sensor and a frequency-differential EIT coupled with a single-cell equivalent circuit-based reconstruction algorithm. The micro-EIT sensor is designed to match single-cell scale and fabricated on a glass substrate by electron beam lithography, which enables high spatial resolution (7 μm electrode width, 40 μm spacing), stable frequency response, and signal-to-noise ratios typically ranging from 50 to 200. The frequency-difference EIT achieves the reconstruction of conductivity distributions of the cytoplasm <em>σ<small><sub>cyt</sub></small></em> and nucleoplasm <em>σ<small><sub>nuc</sub></small></em> through current response analysis based on the equivalent circuit model of single-cell. To evaluate the performance, impedance spectra were measured to reconstruct the intracellular conductivity images in three types of Medical Research Council 5 human lung fibroblast cell lines (MRC-5) with different protein expressions. As a result, <em>σ<small><sub>cyt</sub></small></em> and <em>σ<small><sub>nuc</sub></small></em> of three cell types were successfully reconstructed, which revealed clear differences corresponding to variations in protein expression. The brightfield and fluorescence observation were also performed to verify the EIT results, which demonstrated the reliability of the coordinates and size of the cytoplasm and nucleoplasm. This work represents the first demonstration of non-invasive intracellular conductivity mapping that distinguishes subcellular structures based on electrical properties.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"5 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145183231","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 streamlined lateral flow immunoassay for S. typhimurium using intrinsically multifunctional magnetic nanoprobes for capture, enrichment, and signal amplification.","authors":"Xuechi Yin,Jingxin Hou,Jiayi Zhang,Yumiao Jian,Nosirjon Sattorov,Xinnan Liu,Jianlong Wang,Qingyu Yang,Daohong Zhang,Ibrahim A Darwish","doi":"10.1039/d5lc00437c","DOIUrl":"https://doi.org/10.1039/d5lc00437c","url":null,"abstract":"Salmonella typhimurium (S. typhimurium) is a major foodborne pathogen, posing significant public health risks and leading to substantial economic losses. Traditional lateral flow immunoassay (LFIA) technology for foodborne pathogen detection faces limitations, such as the challenging and costly process of screening paired antibodies. To address these issues, we developed an innovative label-free LFIA utilizing carboxyl-functionalized Fe3O4 nanoparticles for sensitive detection of S. typhimurium. Fe3O4 nanoparticles offer unique properties, including magnetic separation and peroxidase-like catalytic activity, which enhance both bacterial capture and colorimetric signal amplification. The proposed label-free magnetic separation LFIA (LFMS-LFIA) achieved a detection limit of 103 cfu mL-1 for S. typhimurium using catalytic signal amplification, with a sensitivity 10 times greater than that achieved with colorimetric signal alone. This approach demonstrates promising application potential in S. typhimurium contamination detection in samples such as drinking water, eggs, and skim milk. By utilizing probes that integrate efficient binding, magnetic enrichment, and signal amplification capabilities, this Fe3O4-enhanced LFIA represents a major advancement in S. typhimurium detection with high sensitivity and without the need for complex equipment. This work offers a cost-effective and highly sensitive solution in portable devices of food safety and environmental monitoring fields for the on-site detection of pathogens.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"67 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145182807","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lab on a ChipPub Date : 2025-09-25DOI: 10.1039/d5lc00700c
Li-Sin Tu, Tai-Wei Liu, He-Hsuan Hsiao
{"title":"Signal Amplification Using Ab-AuNPs Integrated with LDI-MS Analysis for Diabetes Screening in Urine and Saliva","authors":"Li-Sin Tu, Tai-Wei Liu, He-Hsuan Hsiao","doi":"10.1039/d5lc00700c","DOIUrl":"https://doi.org/10.1039/d5lc00700c","url":null,"abstract":"The global prevalence of diabetes is rising at an alarming rate, making it the third leading cause of death worldwide. This study presented a user-friendly, straightforward, and non-invasive method for screening diabetes. Various antibody-conjugated boronic acid-modified gold nanoparticles (Ab-AuNPs) were prepared, including anti-HbA1c, anti-HBA1, anti-HSA, anti-gHSA, and anti-insulin, to enable the specific recognition of their corresponding antigens in single droplet samples of urine and saliva on nitrocellulose membranes, with subsequent analysis performed using laser desorption/ionization mass spectrometry (LDI-MS). Ab-AuNPs absorbed ultraviolet laser light, leading to the direct desorption and ionization of Au+ ions. This process eliminated the need for an additional organic matrix in matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS), effectively reduced interference from matrix-related ions, and significantly amplified the detection signal of Au+ ions at trace levels for targeted antigens in urine and saliva. The developed method revealed elevated levels of glycated proteins, including glycated hemoglobin (HbA1c) and glycated human serum albumin (gHSA), as well as human serum albumin (HSA), in diabetes patients compared to healthy individuals. In contrast, insulin levels were notably lower in diabetes patients. By analyzing these biomarker changes, we successfully identified the presence of diabetes. The reported method for screening diabetes in biological fluids provides a practical approach and holds significant promise for analyzing other diseases as corresponding biomarkers are discovered and their antibodies are developed and acquired in the future.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"26 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145133534","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lab on a ChipPub Date : 2025-09-25DOI: 10.1039/d5lc00305a
Morgan N. Greenleaf, Gregory L. Damhorst, Eric M. Vogel, Greg S. Martin, Wilbur A. Lam
{"title":"From startup to shutdown: the dramatic rise and fall of the first at-home combo test for flu and COVID-19","authors":"Morgan N. Greenleaf, Gregory L. Damhorst, Eric M. Vogel, Greg S. Martin, Wilbur A. Lam","doi":"10.1039/d5lc00305a","DOIUrl":"https://doi.org/10.1039/d5lc00305a","url":null,"abstract":"This article explores the development and commercialization of Lucira Health's innovative at-home molecular diagnostic test, which detects influenza A or B and SARS-CoV-2. Launched amidst the urgent demand for accessible testing solutions, Lucira's product represented a significant breakthrough, becoming the first over-the-counter combination test authorized by the US Food and Drug Administration (FDA). The narrative tracks Lucira's journey from its origins in microfluidics at the University of California-Berkeley, through development challenges, business success and failure. It also contrasts the distinct motivations and technical challenges of pre-pandemic <em>versus</em> pandemic era diagnostics, emphasizing test-to-treat strategies <em>versus</em> rapid results for containment. Despite early successes, Lucira faced insurmountable regulatory and financial hurdles, culminating in bankruptcy just days before FDA authorization. The case offers critical insights into diagnostics product development, regulatory navigation, product diversification, and strategic risk management in push towards home and point of care diagnostics.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"13 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145133536","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":"sNails: sweat-sensing nails for unobtrusive, wearable microfluidic sweat monitoring from the dorsal distal phalanges.","authors":"Noelle Davis,Pooja Mehta,Amanda Kang,Liam Gillan,Jussi Hiltunen,Ali Javey","doi":"10.1039/d5lc00586h","DOIUrl":"https://doi.org/10.1039/d5lc00586h","url":null,"abstract":"We present a fingernail-mounted microfluidic sweat sensor, the sNail, that leverages the anatomical and mechanical advantages of the nail plate and adjacent dorsal finger skin-regions that are uniquely stiff and stable yet underutilized for wearable biosensing. In contrast to fully soft, skin-mounted sensors, our device exploits the rigidity of the nail to host microfluidics that remain stable under typical daily finger motion. The platform incorporates a hydrogel-filled sweat collection well and stretchable thermoplastic polyurethane (TPU) microfluidics, fabricated via scalable laserjet printing and laser cutting. We demonstrate frequent sweat rate monitoring across multiple fingers and subjects during activities ranging from walking to typing, and capture dynamic changes in sweat rate associated with both physical exertion and psychological stress. This nail-mounted approach enables unobtrusive, informative sweat biosensing, paving the way for broader adoption in everyday settings.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"13 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145127393","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lab on a ChipPub Date : 2025-09-23DOI: 10.1039/d5lc00627a
Kevin Michael Joslin, Sophia Dateshidze, Seung Won Shin, Adam R Abate, Iain Clark
{"title":"Deterministic Cell Pairing with Simultaneous Microfluidic Merging and Sorting of Droplets","authors":"Kevin Michael Joslin, Sophia Dateshidze, Seung Won Shin, Adam R Abate, Iain Clark","doi":"10.1039/d5lc00627a","DOIUrl":"https://doi.org/10.1039/d5lc00627a","url":null,"abstract":"Cell–cell interactions drive immune activation, tissue repair, and stem cell fate, yet there are few methods that can create large numbers of pre-defined cell pairs to study cell crosstalk. Droplet microfluidics allows high-throughput compartmentalization of multiple cells, but random loading results in <1% of droplets containing the desired combinations. Here, we present Pair Isolation by Coalescence and Sorting (PICS), a microfluidic platform that can generate specific cell pairs through droplet merging and sorting (‘merge-sorting’). PICS detects target combinations using fluorescence and triggers simultaneous electrocoalescence and dielectrophoretic sorting. Using fluorescent dye–loaded droplets, we achieved 98.6% purity of merged and sorted droplets. In experiments using cells stained with three distinct dyes, >90% of desired cell pairs were recovered – compared to fewer than 1% when using random Poisson loading. To demonstrate the utility of PICS for extended co-culture studies, we merged cells in an alginate solution with calcium chloride droplets, producing monodisperse alginate hydrogels in which 93.3% of the beads contained target cell pairs that maintained viability over 18 hours. Compared to selective merger, this approach physically isolates desired droplets, eliminating unmerged contaminants and enabling cleaner downstream workflows. PICS allows off-chip pre-incubation of droplets before pairing, the merger of reagents for multi-step assays, and the rapid isolation of desired droplet pairs – capabilities not jointly accessible with existing approaches. In summary, PICS is a flexible platform to enrich specific combinations of droplets, cells, or particles for high-throughput studies of cell crosstalk.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"40 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145127930","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lab on a ChipPub Date : 2025-09-23DOI: 10.1039/d5lc00634a
Eden Dotan, Dana Yagoda-Aharoni, Eli Shapira, Natan T. Shaked
{"title":"Label-Free Imaging Flow Cytometry with Rare Cell Classification using Motion-Sensitive-Triggered Interferometry","authors":"Eden Dotan, Dana Yagoda-Aharoni, Eli Shapira, Natan T. Shaked","doi":"10.1039/d5lc00634a","DOIUrl":"https://doi.org/10.1039/d5lc00634a","url":null,"abstract":"We present a label-free imaging flow cytometry system that integrates a microfluidic chip imaged by a motion-sensitive (event-based) camera and an interferometric-phase-microscopy module using a simple frame-based camera. The event camera captures activity from the flowing cells corresponding to thousands of frames per second and triggers the significantly slower interferometric camera when a rare cell, requiring more sensitive analysis, is detected via a single raw-interferogram analysis, significantly reduicng data volume. The raw interferometric data serves as an input to a deep neural network for rare-cell classification. We demonstrate using this system to detect and grade rare cancer cells in blood, where the event camera is used to rapidly classify between the common white blood cells and the rare cancer cells, and the interferometric camera is used to grade the cancer cell type (primary/metastatic), as a human model for detecting and grading circulating tumor cells in liquid biopsies. This hybrid approach enables efficient data acquisition, rapid processing, and high sensitivity, significantly reducing computational load, and is expected to find various applications in detecting and processing rare cells in imaging flow cytometry.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"51 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145116867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lab on a ChipPub Date : 2025-09-22DOI: 10.1039/d5lc00359h
Sungsu Park, Chanyang Lee, Seokgyu Han, Seulgi Lee, Jaehyun Lee, Sein Kim, Seunggyu Ko, Howon Lee
{"title":"ECM-Integrated Hanging Drop Platform for Spatially Controlled Assessment of Immune Cell Regulated Tumour Invasion","authors":"Sungsu Park, Chanyang Lee, Seokgyu Han, Seulgi Lee, Jaehyun Lee, Sein Kim, Seunggyu Ko, Howon Lee","doi":"10.1039/d5lc00359h","DOIUrl":"https://doi.org/10.1039/d5lc00359h","url":null,"abstract":"The tumour immune microenvironment (TIME) plays a crucial role in tumour progression and metastasis. Although spheroids effectively model tumour invasion by mimicking in vivo 3D structures, their formation and subsequent mixing with the matrix make it difficult to control their position in the 3D matrix, leading to deep embedding and hindering the assessment of immune cell-mediated regulation of invasion. This paper introduces an extracellular matrix (ECM)-integrated hanging drop platform that enables simultaneous spheroid formation and matrix incorporation, allowing precise spatial control and direct assessment of immune cell- mediated regulation of invasion. In the presence of microglia (MG), cancer cells rapidly migrate out of the spheroids through the ECM, demonstrating cancer invasion. The cytotoxic effect of natural killer (NK) cells on glioblastoma multiforme (GBM) spheroids is decreased owing to the inhibition of NK cell infiltration in the presence of MG, highlighting the immunosuppressive nature of the TIME. However, inhibiting STAT3 activation with drugs halts MG-induced immunosuppression and enhances NK cell infiltration. This model enables efficient high-throughput screening and is the first to allow for precise quantification of the effects of the STAT3 inhibitor on tumour invasion, immune cell movement, and behaviour within a physiologically relevant GBM TIME model.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"79 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145116868","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}