Microfluidic Chip Research Articles

CALR frameshift mutation detection in myeloproliferative neoplasms by microfluidic chip analysis

Abstract Background CALR mutation analysis is routinely used to diagnose BCR/ABL1-negative myeloproliferative neoplasms. The 2 most common CALR mutations are a 52–base pair (bp) deletion and a 5-bp insertion, which account for approximately 85% of cases. Methods To evaluate our new microfluidic chip assay, we tested CALR mutant and wild-type specimens that were previously analyzed using conventional methods at a reference laboratory. Samples included EDTA-anticoagulated peripheral blood and bone marrow specimens, air dried bone marrow aspirate smears, and formalin-fixed, paraffin-embedded bone marrow sections. CALR exon 9 was PCR amplified using 2 previously published primer pairs and a third unique primer pair designed for our new assay. Amplicons were sized using microfluidic chip analysis. Results Concordance with the reference method was 100% (42/42). Intra-run and inter-run reproducibility were also 100% (3/3 and 3/3, respectively). The limit of detection was confirmed to be 6% mutant alleles. Conclusion We determined that the microfluidic chip assay to detect CALR exon 9 mutations was acceptable for clinical use. Compared with the conventional method, the microfluidic analysis assay benefits from a streamlined workflow, faster turnaround, and a smaller instrument footprint.

Rapid Fluid Velocity Field Prediction in Microfluidic Mixers via Nine Grid Network Model.

The rapid advancement of artificial intelligence is transforming the computer-aided design of microfluidic chips. As a key component, microfluidic mixers are widely used in bioengineering, chemical experiments, and medical diagnostics due to their efficient mixing capabilities. Traditionally, the simulation of these mixers relies on the finite element method (FEM), which, although effective, presents challenges due to its computational complexity and time-consuming nature. To address this, we propose a nine-grid network (NGN) model theory with a centrally symmetric structure.The NGN uses a symmetric structure similar to a 3 × 3 grid to partition the fluid space to be predicted. Using this theory, we developed and trained an artificial neural network (ANN) to predict the fluid dynamics within microfluidic mixers. This approach significantly reduces the time required for fluid evaluation. In this study, we designed a prototype microfluidic mixer and validated the reliability of our method by comparing it with predictions from traditional FEM software. The results show that our NGN model completes fluid predictions in just 40 s compared to approximately 10 min with FEM, with acceptable error margins. This technology achieves a 15-fold acceleration, greatly reducing the time and cost of microfluidic chip design.

Development of Pyramidal Microwells for Enhanced Cell Spheroid Formation in a Cell-on-Chip Microfluidic System for Cardiac Differentiation of Mouse Embryonic Stem Cells.

Three-dimensional (3D) tissue culture models provide in vivo-like conditions for studying cell physiology. This study aimed to examine the efficiency of pyramidal microwell geometries in microfluidic devices on spheroid formation, cell growth, viability, and differentiation in mouse embryonic stem cells (mESCs). The static culture using the hanging drop (HD) method served as a control. The microfluidic chips were fabricated to have varying pyramidal tip angles, including 66°, 90°, and 106°. From flow simulations, when the tip angle increased, streamline distortion decreased, resulting in more uniform flow and a lower velocity gradient near the spheroids. These findings demonstrate the significant influence of microwell geometry on fluid dynamics. The 90° microwells provide optimal conditions, including uniform flow and reduced shear stress, while maintaining the ability for waste removal, resulting in superior spheroid growth compared to the HD method and other microwell designs. From the experiments, by Day 3, spheroids in the 90° microwells reached approximately 400 µm in diameter which was significantly larger than those in the 66° microwells, 106° microwells, and HD cultures. Brachyury gene expression in the 90° microwells was four times higher than the HD method, indicating enhanced mesodermal differentiation essential for cardiac differentiation. Immunofluorescence staining confirmed cardiomyocyte differentiation. In conclusion, microwell geometry significantly influences 3D cell culture outcomes. The pyramidal microwells with a 90° tip angle proved most effective in promoting spheroid growth and cardiac differentiation of mESC differentiation, providing insights for optimizing microfluidic systems in tissue engineering and regenerative medicine.

Portable microfluidic devices for monitoring antibiotic resistance genes in wastewater.

Antibiotic resistance genes (ARGs) pose serious threats to environmental and public health, and monitoring ARGs in wastewater is a growing need because wastewater is an important source. Microfluidic devices can integrate basic functional units involved in sample assays on a small chip, through the precise control and manipulation of micro/nanofluids in micro/nanoscale spaces, demonstrating the great potential of ARGs detection in wastewater. Here, we (1) summarize the state of the art in microfluidics for recognizing ARGs, (2) determine the strengths and weaknesses of portable microfluidic chips, and (3) assess the potential of portable microfluidic chips to detect ARGs in wastewater. Isothermal nucleic acid amplification and CRISPR/Cas are two commonly used identification elements for the microfluidic detection of ARGs. The former has better sensitivity due to amplification, but false positives due to inappropriate primer design and contamination; the latter has better specificity. The combination of the two can achieve complementarity to a certain extent. Compared with traditional microfluidic chips, low-cost and biocompatible paper-based microfluidics is a very attractive test for ARGs, whose fluid flow in paper does not require external force, but it is weaker in terms of repeatability and high-throughput detection. Due to that only a handful of portable microfluidics detect ARGs in wastewater, fabricating high-throughput microfluidic chips, developing and optimizing recognition techniques for the highly selective and sensitive identification and quantification of a wide range of ARGs in complex wastewater matrices are needed.

Lab-on-a-Chip Devices for Nucleic Acid Analysis in Food Safety.

Lab-on-a-chip (LOC) devices have been developed for nucleic acid analysis by integrating complex laboratory functions onto a miniaturized chip, enabling rapid, cost-effective, and highly sensitive on-site testing. This review examines the application of LOC technology in food safety, specifically in the context of nucleic acid-based analyses for detecting pathogens and contaminants. We focus on microfluidic-based LOC devices that optimize nucleic acid extraction and purification on the chip or amplification and detection processes based on isothermal amplification and polymerase chain reaction. We also explore advancements in integrated LOC devices that combine nucleic acid extraction, amplification, and detection processes within a single chip to minimize sample preparation time and enhance testing accuracy. The review concludes with insights into future trends, particularly the development of portable LOC technologies for rapid and efficient nucleic acid testing in food safety.

Elucidating Extracellular Vesicle Isolation Kinetics via an Integrated Off-Stoichiometry Thiol-Ene and Cyclic Olefin Copolymer Microfluidic Device.

Extracellular vesicles (EVs) are promising biomarkers for diagnosing complex diseases such as cancer and neurodegenerative disorders. Yet, their clinical application is hindered by challenges in isolating cancer-derived EVs efficiently due to their broad size distribution in biological samples. This study introduces a microfluidic device fabricated using off-stoichiometry thiol-ene and cyclic olefin copolymer, addressing the absorption limitations of polydimethylsiloxane (PDMS). The device streamlines a standard laboratory assay into a semi-automated microfluidic chip, integrating sample mixing and magnetic particle separation. Using the microfluidic device, the binding kinetics between EVs and anti-CD9 nanobodies were measured for the first time. Based on the binding kinetics, already after 10 min the EV capture was saturated and comparable to standard laboratory assays, offering a faster alternative to antibody-based immunomagnetic protocols. Furthermore, this study reveals the binding kinetics of EVs to anti-CD9 nanobodies for the first time. Our findings demonstrate the potential of the microfluidic device to enhance clinical diagnostics by offering speed and reducing manual labor without compromising accuracy.

Applications of Nanotechnology for Spatial Omics: Biological Structures and Functions at Nanoscale Resolution.

Spatial omics methods are extensions of traditional histological methods that can illuminate important biomedical mechanisms of physiology and disease by examining the distribution of biomolecules, including nucleic acids, proteins, lipids, and metabolites, at microscale resolution within tissues or individual cells. Since, for some applications, the desired resolution for spatial omics approaches the nanometer scale, classical tools have inherent limitations when applied to spatial omics analyses, and they can measure only a limited number of targets. Nanotechnology applications have been instrumental in overcoming these bottlenecks. When nanometer-level resolution is needed for spatial omics, super resolution microscopy or detection imaging techniques, such as mass spectrometer imaging, are required to generate precise spatial images of target expression. DNA nanostructures are widely used in spatial omics for purposes such as nucleic acid detection, signal amplification, and DNA barcoding for target molecule labeling, underscoring advances in spatial omics. Other properties of nanotechnologies include advanced spatial omics methods, such as microfluidic chips and DNA barcodes. In this review, we describe how nanotechnologies have been applied to the development of spatial transcriptomics, proteomics, metabolomics, epigenomics, and multiomics approaches. We focus on how nanotechnology supports improved resolution and throughput of spatial omics, surpassing traditional techniques. We also summarize future challenges and opportunities for the application of nanotechnology to spatial omics methods.

Detection of circulating tumor cells using a microfluidic chip for diagnostics and therapeutic prediction in mediastinal neuroblastoma.

Circulating tumor cells (CTCs) have served as noninvasive tumor biomarkers in many types of cancer. Here, we detected CTCs in mediastinal neuroblastoma (mNB) patients for use as diagnostic and treatment response predictive biomarkers. We employed a cascaded filter deterministic lateral displacement microfluidic chip (CFD-Chip) to enrich CTCs in peripheral blood from 32 mNB patients and 7 healthy children. CTCs were identified by immunofluorescence staining and integrated neoplastic cell morphology. In total, 66.67% of newly diagnosed mNB patients were positive for CTCs while no CTCs were detected in healthy children. Moreover, CTC count differed significantly across different International Neuroblastoma Staging System, International Neuroblastoma Risk Group staging system, and risk stratifications. CTC count was also significantly higher in children with metastasis than those without metastasis. Additionally, CTC demonstrated a significant difference among patients with different clinical responses to therapy. CTC count decreased or fluctuated at low levels in patients with complete and partial response, compared to considerably increased in patients with stable and progressive diseases.Conclusion: CTCs may serve as non-invasive indicators for mNB diagnosis, staging, and metastasis prediction, and demonstrate promising potential as a liquid biopsy biomarker for the dynamic monitoring of therapeutic efficacy.

Confinement-sensitive volume regulation dynamics via high-speed nuclear morphological measurements

Diverse tissues in vivo present varying degrees of confinement, constriction, and compression to migrating cells in both homeostasis and disease. The nucleus in particular is subjected to external forces by the physical environment during confined migration. While many systems have been developed to induce nuclear deformation and analyze resultant functional changes, much remains unclear about dynamic volume regulation in confinement due to limitations in time resolution and difficulty imaging in PDMS-based microfluidic chips. Standard volumetric measurement relies on confocal microscopy, which suffers from high phototoxicity, slow speed, limited throughput, and artifacts in fast-moving cells. To address this, we developed a form of double fluorescence exclusion microscopy, designed to function at the interface of microchannel-based PDMS sidewalls, that can track cellular and nuclear volume dynamics during confined migration. By verifying the vertical symmetry of nuclei in confinement, we obtained computational estimates of nuclear surface area. We then tracked nuclear volume and surface area under physiological confinement at a time resolution exceeding 30 frames per minute. We find that during self-induced entrance into confinement, the cell rapidly expands its surface area until a threshold is reached, followed by a rapid decrease in nuclear volume. We next used osmotic shock as a tool to alter nuclear volume in confinement, and found that the nuclear response to hypo-osmotic shock in confinement does not follow classical scaling laws, suggesting that the limited expansion potential of the nuclear envelope might be a constraining factor in nuclear volume regulation in confining environments in vivo.

Size-dependent selectivity and quantification on detecting PS nanoplastics particles in a mixed solution with different diameters by using periodic Ag nanocavities SERS substrates with high sensitivity.

Nanoplastic particles (NPPs) have attracted lots of attention due to their toxicity. In this study, a Surface-enhanced Raman scattering (SERS)-based category on selectivity and quantification detecting the polystyrene (PS) NPPs has been presented. Firstly, the size-dependent SERS relationship between the diameter of Ag nanocavities (AgNCAs) and the diameter of the PS NPPs is studied. By continuously dripping the PS NPPs on proposed AgNCAs substrates, AgNCAs exhibit excellent enrichment capability with a promoted limit of detection (LOD) of 0.001 mg/mL. Secondly, thermally evaporated Ag nanoparticles (AgNPs) as an enhancement layer are used to form the AgNPs/PS NPPs/AgNCAs sandwich structure with a SERS enhancement of 300 %. Thirdly, a SERS microfluidic chip constructed by integrating two kinds of pore size (87 nm and 356 nm) AgNCAs is fabricated to selectivity quantifying absolute concentration of the mixed PS NPPs with different diameters in a mixed solution. It shows excellent performance. This novel category proves a good method for identifying plastic nanoparticles and analyzing their size distribution existing in the surroundings indicating good practical applications.

Surface wettability governs brine evaporation and salt precipitation during carbon sequestration in saline aquifers: Microfluidic insights.

Carbon sequestration in deep saline aquifers is a promising strategy for reducing atmospheric CO2 emissions. However, salt precipitation triggered by the evaporation of formation brine into injected supercritical CO2 can cause injectivity and containment issues in near-wellbore regions. Predicting the distribution of precipitated salts and their impact on near-wellbore properties remains challenging. This study investigates the influence of surface wettability on CO2-induced halite precipitation and growth within hydrophilic and hydrophobic microfluidic chips designed to mimic rock-structure porous geometries. A series of high-pressure brine-CO2 flow experiments, direct microscopic observations, and detailed image processing were conducted to explore how substrate wettability affects salt precipitation. The experiments show that wettability markedly controls residual brine relocation, film flow movement, solute consumption, and salt formation. Tracking halite precipitation dynamics revealed distinct crystal formation signatures: hydrophilic chips exhibited irregular, larger aggregation patches, while the hydrophobic network showed more numerous, smaller, and limited aggregations. Large individual crystals were observed in both chips, with a notable dominance in the hydrophobic one. Crystallization dynamics varied, with nucleation and growth occurring earlier, progressing faster, and forming bulkier aggregates in the hydrophilic chip. Despite these differences, the three identified temporal stages of brine evaporation and halite surface coverage were comparable. Spatial analysis along the chips indicated that crystal aggregation properties, such as size and distribution, were position-dependent, with hydrophilic chips exhibiting greater probabilistic variability. These observations underscore the impact of surface wettability on salt precipitation through brine accessibility and capillarity, with implications for mitigating and remediating salt issues in saline aquifers.

Microfluidic chip systems for characterizing glucose-responsive insulin-secreting cells equipped with FailSafe kill-switch

BackgroundPluripotent cell-derived islet replacement therapy offers promise for treating Type 1 diabetes (T1D), but concerns about uncontrolled cell proliferation and tumorigenicity present significant safety challenges. To address the safety concern, this study aims to establish a proof-of-concept for a glucose-responsive, insulin-secreting cell line integrated with a built-in FailSafe kill-switch.MethodWe generated β cell-induced progenitor-like cells (βiPLCs) from primary mouse pancreatic β cells through interrupted reprogramming. Then, we transcriptionally linked our FailSafe (FS) kill-switch, HSV-thymidine kinase (TK), to Cdk1 gene using a CRISPR/Cas9 knock-in strategy, resulting in a FailSafe βiPLC line, designated as FSβiPLCs. Subsequently we evaluated and confirmed the functionality of the drug-inducible kill-switch in FSβiPLCs at different ganciclovir (GCV) concentrations using our PDMS-based transcapillary microfluidic system. Finally, we assessed the functionality of FSβiPLCs by characterizing the dynamics of insulin secretion in response to changes in glucose concentration using our microfluidic perfusion glucose-stimulated insulin secretion (GSIS) assay-on- chip.ResultsThe βiPLCs exhibited Ins1, Pdx1 and Nkx6.1 expression, and glucose responsive insulin secretion, the essential properties of pancreatic beta cells. The βiPLCs were amenable to genome editing which allowed for the insertion of the kill-switch into the 3’UTR of Cdk1, confirmed by PCR genotyping. Our transcapillary microfluidic system confirmed the functionality of the drug-inducible kill-switch in FSβiPLCs, showing an effective cell ablation of dividing cells from a heterogeneous cell population at different ganciclovir (GCV) concentrations. The Ki67 expression assessment further confirmed that slow- or non-dividing cells in the FSβiPLC population were resistant to GCV. Our perfusion glucose-stimulated insulin secretion (GSIS) assay-on-chip revealed that the resistant non-dividing FSβiPLCs exhibited higher levels of insulin secretion and glucose responsiveness compared to their proliferating counterparts.ConclusionsThis study establishes a proof-of-concept for the integration of a FailSafe kill-switch system into a glucose-responsive, insulin-secreting cell line to address the safety concerns in stem cell-derived cell replacement treatment for T1D. The microfluidic systems provided valuable insights into the functionality and safety of these engineered cells, demonstrating the potential of the kill-switch to reduce the risk of tumorigenicity in pluripotent cell-derived insulin-secreting cells.

Retraction Note: Label-free microfluidic chip for segregation and recovery of circulating leukemia cells: clinical applications in acute myeloid leukemia.

Retraction Note: Label-free microfluidic chip for segregation and recovery of circulating leukemia cells: clinical applications in acute myeloid leukemia.

Concurrent Detection of Protein and miRNA at the Single Extracellular Vesicle Level Using a Digital Dual CRISPR-Cas Assay.

The simultaneous detection of proteins and microRNA (miRNA) at the single extracellular vesicle (EV) level shows great promise for precise disease profiling, owing to the heterogeneity and scarcity of tumor-derived EVs. However, a highly reliable method for multiple-target analysis of single EVs remains to be developed. In this study, a digital dual CRISPR-Cas-powered Single EV Evaluation (ddSEE) system was proposed to enable the concurrent detection of surface protein and inner miRNA of EVs at the single-molecule level. By optimizing simultaneous reaction conditions of CRISPR-Cas12a and CRISPR-Cas13a, the surface protein of EVs was detected by Cas12a using antibody-DNA conjugates to transfer the signal of the protein to DNA, while the inner miRNA was analyzed by Cas13a through EV-liposome fusion. A microfluidic chip containing 188,000 microwells was used to convert the CRISPR-Cas system into a digital assay format to enable the absolute quantification of miRNA/protein-positive EVs without bias through fluorescence imaging, which can detect as few as 214 EVs/μL. Finally, a total of 31 blood samples, 21 from breast cancer patients and 10 from healthy donors, were collected and tested, achieving a diagnostic accuracy of 92% in distinguishing patients with breast cancer from healthy donors. With its absolute quantification, ease of use, and multiplexed detection capability, the ddSEE system demonstrates its great potential for both EV research and clinical applications.

On Multicell-Interaction Chip: In Situ Observing the Interactions between the Astrocytes with Lysosomal Dysfunction and BBB Cells.

Lysosomes in astrocytes play vital roles in toxic protein degradation in the brain. Lysosomal dysfunction can lead to abnormal protein deposits, which further induce damage to neurons and the blood-brain barrier (BBB), and thereby affect the interaction between the nervous and vascular systems. Therefore, investigating the interactions between astrocytes with lysosomal dysfunction and BBB cells is of significant importance. However, the lack of effective in vitro models hinders the study of this complex system. Herein, an 8-well arrayed microfence multicell interculture chip (AMMIC) with a hydrophilically optimized surface is introduced for investigating the interactions between astrocytes and BBB cells. Then, a novel lysosome-targeted photosensitizer, IVQ-2Br, is synthesized for inducing controllable oxidative stress damage in the lysosomes of astrocytes. By the combination of the 8-well AMMIC and IVQ-2Br, a model for studying the interactions between astrocytes with lysosomal dysfunction and BBB cells has been constructed. Particularly, severe secondary injuries to BBB cells brought about by oxidative stress, including alterations in cell morphology and activity as well as notable DNA damage, are in situ observed on the 8-well AMMIC. The mediators involved in this oxidative stress injury-mediated intercellular communication are validated to be reactive oxygen species (ROS) and exosomes. This work not only presents an in vitro modeling method for studying cell-cell interactions but also demonstrates the potential of in vitro models constructed through the integration of complex microfluidic chip techniques and photosensitizers for advancing biomedical research.

Microfluidic Chip-based Enrichment and Nucleic Acid Extraction for Quantitative Detection of Mycobacterium Smegmatis in Aerosols.

Tuberculosis (TB) is ranked as the third most prevalent infectious disease globally. Early detection and treatment are crucial for effective management. Conventional diagnostic methods primarily rely on sputum samples, which present challenges in accessibility and have limited accuracy in certain populations such as children, individuals with HIV, and those with extrapulmonary TB. To address the need for point-of-care diagnostics, this study introduces a rapid diagnostic approach for TB using exhaled breath aerosol as a more easily obtainable specimen. Mycobacterium smegmatis, a non-pathogenic bacterium genetically similar to Mycobacterium tuberculosis, is used as a surrogate organism. The method involves the use of microfluidic chips for concentrating and electrolyzing mycobacteria in the aerosol, followed by extracting and quantifying nucleic acids using real-time fluorescence quantitative PCR. Notably, successful enrichment and quantification of bacterial content were achieved even at a minimal bacterial aerosol concentration of 104 CFU/mL. The developed chips are characterized by their cost-effectiveness, ease of use, high bacterial enrichment, efficient nucleic acid extraction, and low detection threshold (4.4 × 10-18 mol/L). This innovative approach offers a promising method for early TB screening and opens avenues for the rapid identification of other aerosol-transmitted diseases.

Additive manufacturing of 3D flow-focusing millifluidics for the production of curable microdroplets.

Microfluidics plays a crucial role in the generation of mono-sized microdroplet emulsions. Traditional glass microfluidic chips typically lack versatility in generating curable droplets of arbitrary liquids due to the inherent hydrophilic nature of glass and to fabrication constraints. To overcome this, we designed a microdroplet generator with 3D flow-focusing capabilities that can be 3D-printed. The chip can handle oil-in-water emulsions despite its lipophilicity. Operating in the jetting regime, the chip exploits the Rayleigh-Plateau instability to enable high throughput. With its versatile design, the chip is capable of producing both single and double emulsions within the same channel. We utilize a thermoset (epoxy-melamine) based system to test its ability to handle curable chemicals and to produce in a post-processing step both solid particles and filled capsules. With a low solvent concentration in the curable material, the present system can encapsulate water-based cores of a wide range of sizes.

Time-resolved cryo-EM (TRCEM) sample preparation using a PDMS-based microfluidic chip assembly.

Design and fabrication of high-performance splitting-and-recombination-based micromixer and planar microsprayer. Protocol for SiO 2 coating on the PDMS surface and fabrication of the microfluidic chip assembly. Preparation of time-resolved cryo-EM sample in the time range of 10 - 1000 ms.Data collection on EM grid covered with droplets from the microsprayer.

Raman spectroscopy with a microfluidic device embedded with plasmonic metasurface.

Metasurfaces offer a powerful tool to realize label-free and highly sensitive Raman spectroscopy. Embedding metasurfaces into microfluidic channels is promising to establish a new characterizing platform for microfluids. In this Letter, we present a highly stable method for improving the Raman scattering intensity of biological microfluids by using a microfluidic chip embedded with a plasmonic metasurface. The embedded metasurface consists of a nanosphere array coated with a silver layer, where the diameter of the nanosphere is ∼100 nm. The Langmuir-Blodgett method and a chemical spraying method were adopted to prepare the nanosphere-array metasurface. In the case of red blood cell measurement, a giant enhancement of Raman spectra intensity is achieved with a metasurface compared to that without a metasurface. Moreover, a two-time enhancement of Raman spectra intensity is obtained with a metasurface under radially polarized beam illumination compared to linearly polarized beam illumination. Furthermore, a microfluidic device embedded with a plasmonic metasurface was applied to monitor the environmental variation of rat red blood cells. Peaks in the range from 2143 cm-1 to 2303 cm-1 arise with the addition of glucose and are still obviously distinguishable when the additive concentration is down to 10-3 M. This indicates high sensitivity to the concentration of glucose mixed with rat red blood cells, which could be further applied to monitor biological cell environments such as glucose concentration, pH, and sodium salt concentration.

On‐Chip Engineered Living Materials as Field‐Deployable Biosensing Laboratories for Multiplexed Detection

AbstractEngineered living materials (ELMs) harness engineered cells to fabricate functional materials with lifelike characteristics, offering unparalleled potential across various fields. Nonetheless, the deployment of ELM‐based biosensors beyond laboratory settings remains challenging. Herin, ELMs are explored as field‐deployable biosensing laboratories on a microfluidic chip (ELMlab‐on‐Chip) for the simultaneous detection of diverse analytes in the field. This approach engages a bottom‐up strategy that includes the molecular engineering of living biosensors, the construction of stimuli‐responsive ELMs, and the fabrication of an integrated biosensing device. Specifically, living biosensors are engineered with fine‐tuned sensitivity and response by designing chimeric receptors and precisely controlling receptor concentration. Integrating ionic and covalent cross‐linking strategies in manufacturing ELMs ensures good substance permeability and mechanical robustness. Moreover, a microfluidic chip is devised tailored for the orthogonally stimuli‐responsive ELMs, creating a spatially encoded sensor array with the output detected by a miniaturized smartphone‐based detection device. The integrated ELMlab‐on‐Chip platform has demonstrated its potential in the simultaneous analysis of multiple chemicals from a single environmental sample under field conditions, offering an effective strategy to expedite the real‐world application of living materials.