Microfluidic Platform Research Articles

Diffusiophoresis promotes phase separation and transport of biomolecular condensates

The internal microenvironment of a living cell is heterogeneous and comprises a multitude of organelles with distinct biochemistry. Amongst them are biomolecular condensates, which are membrane-less, phase-separated compartments enriched in system-specific proteins and nucleic acids. The heterogeneity of the cell engenders the presence of multiple spatiotemporal gradients in chemistry, charge, concentration, temperature, and pressure. Such thermodynamic gradients can lead to non-equilibrium driving forces for the formation and transport of biomolecular condensates. Here, we report how ion gradients impact the transport processes of biomolecular condensates on the mesoscale and biomolecules on the microscale. Utilizing a microfluidic platform, we demonstrate that the presence of ion concentration gradients can accelerate the transport of biomolecules, including nucleic acids and proteins, via diffusiophoresis. This hydrodynamic transport process allows localized enrichment of biomolecules, thereby promoting the location-specific formation of biomolecular condensates via phase separation. The ion gradients further impart directional motility of condensates, allowing them to exhibit enhanced diffusion along the gradient. Coupled with a reentrant phase behavior, the gradient-induced enhanced motility leads to a dynamical redistribution of condensates that ultimately extends their lifetime. Together, our results demonstrate diffusiophoresis as a non-equilibrium thermodynamic force that governs the formation and transport of biomolecular condensates.

Rapid flowing cells localization enabled by spatiotemporal manipulation of their holographic patterns.

Lab-on-a-Chip microfluidic devices present an innovative and cost-effective platform in the current trend of miniaturization and simplification of imaging flow cytometry; they are excellent candidates for high-throughput single-cell analysis. In such microfluidic platforms, cell tracking becomes a fundamental tool for investigating biophysical processes, from intracellular dynamics to the characterization of cell motility and migration. However, high-throughput and long-term cell tracking puts a high demand on the consumption of computing resources. Here, we propose a novel strategy to achieve rapid 3D cell localizations along the microfluidic channel. This method is based on the spatiotemporal manipulation of recorded holographic interference fringes, and it allows fast and precise localization of cells without performing complete holographic reconstruction. Conventional holographic tracking is typically based on the phase contrast obtained by decoupling the calculation of optical axial and transverse coordinates. Computing time and resource consumption may increase because all the frames need to be calculated in the Fourier domain. In our proposed method, the 2D transverse positions are directly located by morphological calculation based on the hologram. The complex-amplitude wavefronts are directly reconstructed by spatiotemporal phase shifting to calculate the axial position by the refocusing criterion. Only spatial calculation is considered in the proposed method. We demonstrate that the computational time of transverse tracking is only one-tenth of the conventional method, while the total computational time of the proposed method decreases up to 54% with respect to the conventional approach. The proposed approach can open the route for analyzing flow cytometry in quantitative phase microscopy assays.

Recent advances in portable devices for environmental monitoring applications.

Environmental pollution remains a major societal problem, leading to serious impacts on living organisms including humans. Human activities such as civilization, urbanization, and industrialization are major causes of pollution. Imposing stricter rules helps control environmental pollutant levels, creating a need for reliable pollutant monitoring in air, water, and soil. The application of traditional analytical techniques is limited in low-resource areas because they are sophisticated, expensive, and bulky. With the development of biosensors and microfluidics technology, environmental monitoring has significantly improved the analysis time, low cost, portability, and ease of use. This review discusses the fundamentals of portable devices, including microfluidics and biosensors, for environmental control. Recently, publications reviewing microfluidics and biosensor device applications have increased more than tenfold, showing the potential of emerging novel approaches for environmental monitoring. Strategies for enzyme-, immunoassay-, and molecular-based analyte sensing are discussed based on their mechanisms and applications. Microfluidic and biosensor platforms for detecting major pollutants, including metal ions, pathogens, pesticides, and antibiotic residues, are reviewed based on their working principles, advantages, and disadvantages. Challenges and future trends for the device design and fabrication process to improve performance are discussed. Miniaturization, low cost, selectivity, sensitivity, high automation, and savings in samples and reagents make the devices ideal alternatives for in-field detection, especially in low-resource areas. However, their operation with complicated environmental samples requires further research to improve the specificity and sensitivity. Although there is a wide range of devices available for environmental applications, their implementation in real-world situations is limited. This study provides insights into existing issues that can be used as references and a comparative analysis for future studies and applications.

Determination of Quercetin in Iraqi Green Walnut Husk via Green Ionic Liquid-Based Digestion Extraction and Gel Filtration Procedures-Spectrophotometry and Microfluidic Paper-Based Analytical Platform Detection

في هذه الدراسة تم استخلاص ،عزل وتقدير الكيورستين الموجود في قشور الجوز العراقي باستخدام طريقة جديدة تعتمد على الاستخلاص بطريقة الهضم المبنية على استخدام السوائل الايونية و طريقة الترشيح بالجل للفصل المقترنة مع المطيافية الضوئية للتقدير ومن ثم مقارنتها مع تقنية جديدة تسمى تقنية السوائل المايكروية الورقية مستهدفة بناء طريقة اقتصادية، صديقة للبيئة وخضراء. ولقد تم اختيار 1 بيوتيل-3-مثيل ايمادزوليم كلورايد كمذيب اخضر بديل للمذيبات التقليدية بسبب خصائصة الذوبانية .ولقد اظهرت النتائج ان 100 ملغرام من القشورتحتاج الى 15 مل من المذيب و30 دقيقة من الهضم عند درجة75°C كظروف مثلى لاعطاء افضل ناتج للمستخلص بطريقة الهضم مقارنه مع طرق الاستخلاص الاخرى .تحت هذه الظروف المثلى تم عزل الكيورستين باستخدام طريقة الترشيح بالجل وتم التاكد من المركب المعزول باستخدام تقنية كروموتوغرافيا السائل عالية الاداء و اشعة فورية تحت الحمراء .تم تقدير الكيورستين المعزول من المستخلص لونيا ( طريقة كلوريد الالمنيوم) باستخدام طريقتي المطيافية الضوئية و وتقنية السوائل المايكروية الورقية معطيا الخطية لمنحنيات المعايرة لكلتا الطريقيتين بين 10-100 مليغرام / لتر مع معامل ارتباط0.9369 و 0.7917 لكلتا الطريقتين. ومن ثم تم تقدير الفعالية المضادة للاكسدة باستخجام طريقة قوة الاختزال للمركب المعزول مظهرة فعالية مضادة للاكسدة جيدة و بين تحليل التباين (ANOVA) عدم وجود فرق كبير بين تقنية المطيافية الضوئية وتقنية السوائل المايكروية الورقية. كخلاصة يمكن اعتبار المذيب الايوني BMIMCLمذيبا امنا ومناسبا لاستخلاص المركبات النباتية ذات الخواص الطبية وكذلك فان تقنية السوائل المايكروية الورقية اثبتت مقدرتها على التحليل بكفاءة مساوية الى طرق التحليل التقليدية بالاضافة الى خصائصها الخضراء.

Single-cell multi-modal chromatin profiles revealing epigenetic regulations of cells in hepatocellular carcinoma.

Various epigenetic regulations systematically govern gene expression in cells involving various biological processes. Dysregulation of the epigenome leads to aberrant transcriptional programs and subsequently results in diseases, such as cancer. Therefore, comprehensive profiling epigenomics is essential for exploring the mechanisms underlying gene expression regulation during development and disease. In this study, we developed single-cell chromatin proteins and accessibility tagmentation (scCPA-Tag), a multi-modal single-cell epigenetic profile capturing technique based on barcoded Tn5 transposases and a droplet microfluidics platform. scCPA-Tag enables the simultaneous capture of DNA profiles of histone modification and chromatin accessibility in the same cell. By applying scCPA-Tag to K562 cells and a hepatocellular carcinoma (HCC) sample, we found that the silence of several chromatin-accessible genes can be attributed to lysine-27-trimethylation of the histone H3 tail (H3K27me3) modification. We characterized the epigenetic features of the tumour cells and different immune cell types in the HCC tumour tissue by scCPA-Tag. Besides, a tumour cell subtype (C2) with more aggressive features was identified and characterized by high chromatin accessibility and a lower abundance of H3K27me3 on tumour-promoting genes. Our multi-modal scCPA-Tag provides a comprehensive approach for exploring the epigenetic landscapes of heterogeneous cell types and revealing the mechanisms of gene expression regulation during developmental and pathological processes at the single-cell level. scCPA-Tag offers a highly efficient and high throughput technique to simultaneously profile histone modification and chromatin accessibility within a single cell. scCPA-Tag enables to uncover multiple epigenetic modification features of cellular compositions within tumor tissues. scCPA-Tag facilitates the exploration of the epigenetic landscapes of heterogeneous cell types and provides the mechanisms governing gene expression regulation.

Environmental modulators of algae-bacteria interactions at scale

Interactions between photosynthetic and heterotrophic microbes play a key role in global primary production. Understanding phototroph-heterotroph interactions remains challenging because these microbes reside in chemically complex environments. Here, we leverage a massively parallel droplet microfluidic platform that enables us to interrogate interactions between photosynthetic algae and heterotrophic bacteria in >100,000 communities across ∼525 environmental conditions with varying pH, carbon availability, and phosphorus availability. By developing a statistical framework to dissect interactions in this complex dataset, we reveal that the dependence of algae-bacteria interactions on nutrient availability is strongly modulated by pH and buffering capacity. Furthermore, we show that the chemical identity of the available organic carbon source controls how pH, buffering capacity, and nutrient availability modulate algae-bacteria interactions. Our study reveals the previously underappreciated role of pH in modulating phototroph-heterotroph interactions and provides a framework for thinking about interactions between phototrophs and heterotrophs in more natural contexts.

Emergence of Microfluidic Platforms as Point of Care Diagnostics for the Detection of Plant and Foodborne Pathogens

Emergence of Microfluidic Platforms as Point of Care Diagnostics for the Detection of Plant and Foodborne Pathogens

Integrating Vascular Phenotypic and Proteomic Analysis in an Open Microfluidic Platform.

This research introduces a vascular phenotypic and proteomic analysis (VPT) platform designed to perform high-throughput experiments on vascular development. The VPT platform utilizes an open-channel configuration that facilitates angiogenesis by precise alignment of endothelial cells, allowing for a 3D morphological examination and protein analysis. We study the effects of antiangiogenic agents─bevacizumab, ramucirumab, cabozantinib, regorafenib, wortmannin, chloroquine, and paclitaxel─on cytoskeletal integrity and angiogenic sprouting, observing an approximately 50% reduction in sprouting at higher drug concentrations. Precise LC-MS/MS analyses reveal global protein expression changes in response to four of these drugs, providing insights into the signaling pathways related to the cell cycle, cytoskeleton, cellular senescence, and angiogenesis. Our findings emphasize the intricate relationship between cytoskeletal alterations and angiogenic responses, underlining the significance of integrating morphological and proteomic data for a comprehensive understanding of angiogenesis. The VPT platform not only advances our understanding of drug impacts on vascular biology but also offers a versatile tool for analyzing proteome and morphological features across various models beyond blood vessels.

Controllable preparation and performance study of DAP-4/CuO high-energy microspheres

To enhance the dispersion uniformity of DAP-4/CuO composite fuel and improve the stability and consistency of the entire charge, particularly in small doses, DAP-4/CuO composite microspheres were prepared using a self-built coaxial microfluidic platform. This addressed the issue of uneven dispersion of DAP-4 and CuO, resulting in excellent dispersion of DAP-4/CuO composite microspheres. Furthermore, the impact of CuO content on DAP-4 microspheres was investigated. The samples were analyzed using scanning electron microscopy, laser confocal microscopy, differential scanning calorimetry, high-speed photography, and infrared thermal imaging. The findings revealed that when 1 % NC was added to the binder, FN-5 components were evenly dispersed. Microspheres produced through microfluidic technology exhibited a narrow particle size distribution which significantly improved sample dispersion and enhanced charging process safety. The addition of CuO notably accelerated the thermal decomposition process of DAP-4 microspheres and substantially increased sample burning rate. However, when CuO content exceeded 10 %, further increases did not continue to raise burning rate but led to decreased flame temperature. This study provides a reference for relevant workers.

Microdroplet-SERS platform for single cell-secreted VEGF and extracellular pH analysis in oxidative stress event

Although oxidative stress processes based on reactive oxygen species have been the research focus, analyzing this phenomenon at the single-cell level is challenging. A microfluidic platform combined with highly sensitive surface-enhanced Raman scattering (SERS) technology was constructed. The secretion of the vascular endothelial growth factor (VEGF) and the extracellular microenvironmental pH fluctuations of a single cell during the oxidative stress process were assessed and analyzed. The immune sandwich structure formed between the capture probe and the reporter probe with the linkage of secreted VEGF was built above the probed cell surface in each drop. The self-driven collection behavior of the magnetic bead-based capture probe toward the cell surface significantly amplified the SERS signals of the reporter probe, thus improving the sensitivity and accuracy of detection. 4-Mercaptopyridine (4-Mpy) responding sensitively to pH was applied to label the reporter probe, which endows the extracellular microenvironmental pH sensing during oxidative stress events according to relative SERS intensity. Combined with machine learning to analyze the spectral characteristics, the distinction between normal cells and different types of tumor cells was finally realized. This study makes full use of the advantages of machine learning to process spectral data to explore deep information and provides an auxiliary tool for diagnosing cancer and other medical conditions in the future.

Micro container made of levitated liquid bead

Liquid bead, an emerging digital microfluidic platform offers a wide range of applications in research and industry. Making liquid beads using acoustic levitation technology allows for the fabrication of micro container. Extensive insight into levitated liquid beads is essential for determining their performance. Firstly, this paper unfolds the unique interface of the levitated liquid bead, which has thin membrane of shell material on top and thus serves as a micro container. Next, we characterised the puncture toughness of liquid beads to assess the capability of releasing the liquid content. Liquid bead with larger corresponding shell volume and lower core volume recorded the highest resistance against needle penetration. Finally, we observed the thermal behaviour of liquid beads at elevated temperature. The liquid beads showed no morphological change over 10-minutes period at 50 °C. At 65 °C, liquid bead exceeded the critical temperature threshold, which resulted in breaking the thin membrane and releasing the liquid from the core.

Real-Time Observation of Protein Aggregation at Liquid-Liquid Interfaces in a Microfluidic Device.

A droplet microfluidic device to capture in real-time protein aggregation at liquid-liquid interfaces is described. In contrast to conventional methods, typically characterized by a lag time between the application of interfacial stress and the measurement of protein aggregation, here protein adsorption, the formation of a viscoelastic protein layer, aggregation, and shedding of protein particles into solution is simultaneously monitored. The device is applied to analyze the stability of antibodyformulations over a wide range of concentrations (1-180mgmL-1) at the silicone oil (SO)-water interface under controlled mechanical deformation. The adsorption onto oil droplets induces the formation of a viscoelastic protein layer on a subsecond timescale, which progressively restricts the relaxation of the droplets within the chip. Upon mechanical rupture, the protein layer releases particles in solution. The rate of particle formation increases strongly with concentration, similar to the bulk viscosity. Concentrations above 120mgmL-1 lead to aggregation in seconds and drastically decrease the mechanical perturbations required to shed protein particles in solution. These results are important for the development of formulations at high-protein concentrations (>100mgmL-1) and indicate that particular attention should be given to interface-induced particle formation in this concentration range. In this context, low-volume microfluidic platforms allow the assessment of protein physical instabilities early in development and represent attractive tools to optimize antibody stability and formulation design consuming limited amounts ofmaterial.

Droplet-based microfluidics for engineering shape-controlled hydrogels with stiffness gradient

Current biofabrication strategies are limited in their ability to replicate native shape-to-function relationships, that are dependent on adequate biomimicry of shape, size and spatial heterogeneity, within cell-laden hydrogels. In this study, a diffusion-based microfluidics platform is presented that meets these needs in a two-step process. In the first step, a hydrogel-precursor solution is dispersed into a continuous oil phase within the microfluidics tubing. By adjusting the dispersed and oil phase flow rates, the physical architecture of hydrogel-precursor phases can be adjusted to generate spherical and plug-like structures, as well as continuous meter-long hydrogel-precursor phases (up to 1.75 m). The second step involves the controlled introduction a small molecule-containing aqueous phase through a T-shaped tube connector to enable controlled small molecule diffusion across the interface of the aqueous phase and hydrogel-precursor. Application of this system is demonstrated by diffusing co-initiator sodium persulfate (SPS) into hydrogel-precursor solutions, where the controlled SPS diffusion into the hydrogel-precursor and subsequent photo-polymerization allows for the formation of unique radial stiffness patterns across the shape- and size-controlled hydrogels, as well as allowing the formation of hollow hydrogels with controllable internal architectures. Mesenchymal stromal cells are successfully encapsulated within hollow hydrogels and hydrogels containing radial stiffness gradient. The cells are observed to respond to the microscale spatial heterogeneity as evidenced by increased cell elongation in softer core regions of the hydrogel as compared to the peripheral stiffer hydrogel regions, as well as stiffness-dependent nuclear accumulation of the yes-associated protein mechano-regulator. Finally, breast cancer cells are found to phenotypically switch in response to stiffness gradients, causing a shift in their ability to aggregate, which may have implications for metastasis. The diffusion-based microfluidics will mimic native shape-to-function relationship and provides a platform to further study the roles of micro- and macroscale architectural features that exist within native tissues.

High-throughput affinity measurements of direct interactions between activation domains and co-activators.

Sequence-specific activation by transcription factors is essential for gene regulation1,2. Key to this are activation domains, which often fall within disordered regions of transcription factors3,4 and recruit co-activators to initiate transcription5. These interactions are difficult to characterize via most experimental techniques because they are typically weak and transient6,7. Consequently, we know very little about whether these interactions are promiscuous or specific, the mechanisms of binding, and how these interactions tune the strength of gene activation. To address these questions, we developed a microfluidic platform for expression and purification of hundreds of activation domains in parallel followed by direct measurement of co-activator binding affinities (STAMMPPING, for Simultaneous Trapping of Affinity Measurements via a Microfluidic Protein-Protein INteraction Generator). By applying STAMMPPING to quantify direct interactions between eight co-activators and 204 human activation domains (>1,500 K ds), we provide the first quantitative map of these interactions and reveal 334 novel binding pairs. We find that the metazoan-specific co-activator P300 directly binds >100 activation domains, potentially explaining its widespread recruitment across the genome to influence transcriptional activation. Despite sharing similar molecular properties (e.g. enrichment of negative and hydrophobic residues), activation domains utilize distinct biophysical properties to recruit certain co-activator domains. Co-activator domain affinity and occupancy are well-predicted by analytical models that account for multivalency, and in vitro affinities quantitatively predict activation in cells with an ultrasensitive response. Not only do our results demonstrate the ability to measure affinities between even weak protein-protein interactions in high throughput, but they also provide a necessary resource of over 1,500 activation domain/co-activator affinities which lays the foundation for understanding the molecular basis of transcriptional activation.

Next-generation 3D tumor modeling: A microfluidic platform with biocompatible red carbon dots for live cell imaging in co-cultured elongated spheroid tumor model

Co-culture spheroids mimic tumor architecture more accurately than traditional 2D cell cultures, but non-invasive, long-term tracking of live cells within these 3D models remains a challenge. This study addresses this critical need by developing a novel approach for live cell imaging in U-87/HUF co-culture spheroids. We introduce water-soluble, biocompatible red carbon dots (R-CDs) exhibiting exceptional stability and brightness (21% quantum yield) specifically designed for imaging within these 3D models. Furthermore, we designed a microfluidic chip with ellipsoid-shaped microwells to efficiently generate two distinct co-culture spheroid types: direct mixing and core-shell. R-CDs enabled non-invasive tracking of U-87 cancer cell location within these 3D models demonstrating their efficacy for long-term monitoring of live cells in cancer research. This R-CD and microfluidic technology has the potential to accelerate cancer drug discovery by enabling live cell studies in 3D tumor models.

Artificial intelligence-based droplet size prediction for microfluidic system

PurposeThis study aims to introduce a vision-based model to generate droplets with auto-tuned parameters. The model can auto-adjust the inherent uncertainties and errors involved with the fabrication and operating parameters in microfluidic platform, attaining precise size and frequency of droplet generation.Design/methodology/approachThe photolithography method is utilized to prepare the microfluidic devices used in this study, and various experiments are conducted at various flow-rate and viscosity ratios. Data for droplet shape is collected to train the artificial intelligence (AI) models.FindingsGrowth phase of droplets demonstrated a unique spring back effect in droplet size. The fully developed droplet sizes in the microchannel were modeled using least absolute shrinkage and selection operators (LASSO) regression model, Gaussian support vector machine (SVM), long short term memory (LSTM) and deep neural network models. Mean absolute percentage error (MAPE) of 0.05 and R2 = 0.93 were obtained with a deep neural network model on untrained flow data. The shape parameters of the droplets are affected by several uncontrolled parameters. These parameters are instinctively captured in the model.Originality/valueExperimental data set is generated for varying viscosity values and flow rates. The variation of flow rate of continuous phase is observed here instead of dispersed phase. An automated computation routine is developed to read the droplet shape parameters considering the transient growth phase of droplets. The droplet size data is used to build and compare various AI models for predicting droplet sizes. A predictive model is developed, which is ready for automated closed loop control of the droplet generation.

A Droplet-Based Microfluidic Platform for High-Throughput Culturing of Yeast Cells in Various Conditions.

Yeast plays a significant role in a variety of fields. In particular, it is extensively used as a model organism in genetics and cellular biology studies, and is employed in the production of vaccines, pharmaceuticals, and biofuels. Traditional "bulk"-based studies on yeast growth often overlook cellular variability, emphasizing the need for single-cell analysis. Micro-droplets, tiny liquid droplets with high surface-area-to-volume ratios, offer a promising platform for investigating single or a small number of cells, allowing precise control and monitoring of individual cell behaviors. Microfluidic devices, which facilitate the generation of micro-droplets, are advantageous due to their reduced volume requirements and ability to mimic in vivo micro-environments. This study introduces a custom-designed microfluidic device to encapsulate yeasts in micro-droplets under various conditions in a parallel manner. The results reveal that optimal glucose concentrations promoted yeast growth while cycloheximide and Cu2+ ions inhibited it. This platform enhances yeast cultivation strategies and holds potential for high-throughput single-cell investigations in more complex organisms.

Gene expression-phenotype association study reveals the dual role of TNF-α/TNFR1 signaling axis in confined breast cancer cell migration

AimsWhile enhanced tumor cell migration is a key process in the tumor dissemination, mechanistic insights into causal relationships between tumor cells and mechanical confinement are still limited. Here we combine the use of microfluidic platforms to characterize confined cell migration with genomic tools to systematically unravel the global signaling landscape associated with the migratory phenotype of breast cancer (BC) cells. Meterials and methodsThe spontaneous migration capacity of seven BC cell lines was evaluated in 3D microfluidic devices and their migration capacity was correlated with publicly available molecular signatures. The role of identified signaling pathways on regulating BC migration capacity was determined by receptor stimulation through ligand binding or inhibition through siRNA silencing. Downstream effects on cell migration were evaluated in microfluidic devices, while the molecular changes were monitored by RT-qPCR. Key findingsExpression of 715 genes was correlated with BC cells migratory phenotype, revealing TNF-α as one of the top upstream regulators. Signal transduction experiments revealed that TNF-α stimulates the confined migration of triple negative, mesenchymal-like BC cells that are also characterized by high TNFR1 expression, but inhibits the migration of epithelial-like cells with low TNFR1 expression. TNFR1 was strongly associated with the migration capacity and triple-negative, mesenchymal phenotype. Downstream of TNF/TNFR1 signaling, transcriptional regulation of NFKB seems to be important in driving cell migration in confined spaces. SignificanceTNF-α/TNFR1 signaling axis reveals as a key player in driving BC cells confined migration, emerging as a promising therapeutic strategy in targeting dissemination and metastasis of triple negative, mesenchymal BC cells.

Exploration of Curvature and Stiffness Dual-Regulated Breast Cancer Cell Motility by a Motor-Clutch Model and Cell Traction Force Characterization.

The migration of breast cancer cells is the main cause of death and significantly regulated by physical factors of the extracellular matrix (ECM). To be specific, the curvature and stiffness of the ECM were discovered to effectively guide cell migration in velocity and direction. However, it is not clear what the extent of effect is when these dual-physical factors regulate cell migration. Moreover, the mechanobiology mechanism of breast cancer cell migration in the molecular level and analysis of cell traction force (CTF) are also important, but there is a lack of systematic investigation. Therefore, we employed a microfluidic platform to construct hydrogel microspheres with an independently adjustable curvature and stiffness as a three-dimensional substrate for breast cancer cell migration. We found that the cell migration velocity was negatively correlated to curvature and positively correlated to stiffness. In addition, curvature was investigated to influence the focal adhesion expression as well as the assignment of F-actin at the molecular level. Further, with the help of a motor-clutch mathematical model and hydrogel microsphere stress sensors, it was concluded that cells perceived physical factors (curvature and stiffness) to cause changes in CTF, which ultimately regulated cell motility. In summary, we employed a theoretical model (motor-clutch) and experimental strategy (stress sensors) to understand the mechanism of curvature and stiffness regulating breast cancer cell motility. These results provide evidence of force driven cancer cell migration by ECM physical factors and explain the mechanism from the perspective of mechanobiology.

D4-Chip Reveals Impaired T Cell Function in Sepsis: Insights from Plasma Microenvironment Analysis and Mitochondrial Targeted Therapy.

Sepsis, a systemic inflammation syndrome initiated by infection, poses significant challenges due to its intricate pathophysiology. T cells play a crucial role in combating infections during sepsis. Despite previous observations indicating T cell dysfunction in sepsis, reliable in-vitro detection methods were lacking, and the factors influencing these impairments remained unclear. We developed a novel method using the D4-Chip to assess sepsis T cell migration function. This microfluidic platform enabled precise analysis of migration function under controlled conditions. Additionally, We explored the impact of the plasma microenvironment on T cell behavior, along with the redox environment in sepsis, and assessed the potential efficacy of Mitoquinone mesylate (MitoQ), a mitochondrial-targeted drug. Our findings revealed impaired migration function in sepsis T cells compared to healthy controls. Interestingly, sepsis plasma enhanced the migration of healthy T cells, yet incubation with healthy plasma did not fully restore migration impairments in sepsis T cells. Subsequent investigations uncovered a significant increase in NADH/NAD+ levels in sepsis T cells, with healthy T cells exposed to various sepsis plasma conditions also showing elevated NADH/NAD+ levels. Importantly, MitoQ normalized abnormal intracellular NADH/NAD+ levels and enhanced the migration ability of T cells. Short-term incubation with sepsis plasma does not directly inhibit T cell migration but instead affects T cell function by disrupting the intracellular redox environment. Improving the intracellular redox environment of sepsis patients contributes to restoring impaired migration and proliferation, with MitoQ demonstrating therapeutic potential.