Gradient Hydrogels Spatially Trapped Optical Cell Profiling for Quantitative Blood Cellular Osmotic Analysis.

The quantitative exploration of cellular osmotic responses and a thorough analysis of osmotic pressure-responsive cellular behaviors are poised to offer novel clinical insights into current research. This underscores a paradigm shift in the long-standing approach of colorimetric measurements triggered by red cell lysis. In this study, we engineered a purpose-driven optofluidic platform to facilitate the goal. Specifically, creating photocurable hydrogel traps surmounts a persistent challenge─optical signal interference from fluid disturbances. This achievement ensures a stable spatial phase of cells and the acquisition of optical signals for accurate osmotic response analysis at the single-cell level. Leveraging a multigradient microfluidic system, we constructed gradient osmotic hydrogel traps and developed an imaging recognition algorithm, empowering comprehensive analysis of cellular behaviors. Notably, this system has successfully and precisely analyzed individual and clustered cellular responses within the osmotic dimension. Prospective clinical testing has further substantiated its feasibility and performance in that it demonstrates an accuracy of 92% in discriminating complete hemolysis values (n = 25) and 100% in identifying initial hemolysis values (n = 25). Foreseeably, this strategy should promise to advance osmotic pressure-related cellular response analysis, benefiting further investigation and diagnosis of related blood diseases, blood quality, drug development, etc.

Heterogeneous tissue construction by on-demand bubble-assisted acoustic patterning.

Highly heterogeneous structures are closely related to the realization of the tissue functions of living organisms. However, precisely controlling the assembly of heterogeneous structures is still a crucial challenge. This work presents an on-demand bubble-assisted acoustic method for active cell patterning to achieve high-precision heterogeneous structures. Active cell patterning is achieved by the combined effect of acoustic radiation forces and microstreaming around oscillating bubble arrays. On-demand bubble arrays allow flexible construction of cell patterns with a precision of up to 45 µm. As a typical example, the in vitro model of hepatic lobules, composed of patterned endothelial cells and hepatic parenchymal cells, was constructed and cultured for 5 days. The good performance of urea and albumin secretion, enzymatic activity and good proliferation of both cells prove the feasibility of this technique. Overall, this bubble-assisted acoustic approach provides a simple and efficient strategy for on-demand large-area tissue construction, with considerable potential for different tissue model fabrication.

Magnetically Actuated Hydrogel Stamping-Assisted Cellular Mechanical Analyzer for Stored Blood Quality Detection.

Cellular mechanical property analysis reflecting the physiological and pathological states of cells plays a crucial role in assessing the quality of stored blood. However, its complex equipment needs, operation difficulty, and clogging issues hinder automated and rapid biomechanical testing. Here, we propose a promising biosensor assisted by magnetically actuated hydrogel stamping to fulfill it. The flexible magnetic actuator triggers the collective deformation of multiple cells in the light-cured hydrogel, and it allows for on-demand bioforce stimulation with the advantages of portability, cost-effectiveness, and simplicity of operation. The magnetically manipulated cell deformation processes are captured by the integrated miniaturized optical imaging system, and the cellular mechanical property parameters are extracted from the captured images for real-time analysis and intelligent sensing. In this work, 30 clinical blood samples with different storage durations (<14 days and >14 days) were tested. A deviation of 3.3% in the differentiation of blood storage durations by this system compared to physician annotation demonstrated its feasibility. This system should broaden the application of cellular mechanical assays in diverse clinical settings.

Valve-Adjustable Optofluidic Bio-Imaging Platform for Progressive Stenosis Investigation.

The clinical evidence has proven that valvular stenosis is closely related to many vascular diseases, which attracts great academic attention to the corresponding pathological mechanisms. The investigation is expected to benefit from the further development of an in vitro model that is tunable for bio-mimicking progressive valvular stenosis and enables accurate optical recognition in complex blood flow. Here, we develop a valve-adjustable optofluidic bio-imaging recognition platform to fulfill it. Specifically, the bionic valve was designed with in situ soft membrane, and the internal air-pressure chamber could be regulated from the inside out to bio-mimic progressive valvular stenosis. The developed imaging algorithm enhances the recognition of optical details in blood flow imaging and allows for quantitative analysis. In a prospective clinical study, we examined the effect of progressive valvular stenosis on hemodynamics within the typical physiological range of veins by this way, where the inhomogeneity and local enhancement effect in the altered blood flow field were precisely described and the optical differences were quantified. The effectiveness and consistency of the results were further validated through statistical analysis. In addition, we tested it on fluorescence and noticed its good performance in fluorescent tracing of the clotting process. In virtue of theses merits, this system should be able to contribute to mechanism investigation, pharmaceutical development, and therapeutics of valvular stenosis-related diseases.

Confinement-enhanced microalgal individuals biosensing for digital atrazine assay.

Microalgal sensors are widely recognized for their high sensitivity, accessibility, and low cost. However, the current dilemma of motion-induced spatial phase changes and concentration-related multiple scattering interferes with induced test instability and limited sensitivity, which has hindered their practical applications. Here, a differentiated strategy, named confinement-enhanced microalgal biosensing (C-EMB), is developed and proposed to pave the way. The in-situ printed microgel trap is designed to confine Chlamydomonas reinhardtii individuals, stabilizing their spatial phase. The microgel trap arrays are introduced to eliminate the multiple scattering of microalgae, breaking the existing effective concentration in traditional microalgal sensing and enabling sensitive assays. The integration with lab-on-a-chip technology and a developed digital imaging algorithm empower portable and automated detection. With this system, a microalgae analyzer is developed for atrazine detection, featuring a linear range of 0.04-100 µg/L. We assess the system's performance through practical atrazine assays on commercial food, using a double-blind test against a standard instrument. Our results demonstrate the good accuracy and test stability of this system with the mean bias atrazine detection in corn and sugarcane juice samples (SD) were 1.661 µg/L (3.122 µg/L) and 3.144 µg/L (4.125 µg/L), respectively. This method provides a new paradigm of microalgal sensors and should advance the further applications of microalgal sensors in commercial and practical settings.

A microfluidic hemostatic diagnostics platform: Harnessing coagulation-induced adaptive-bubble behavioral perception.

Clinical viscoelastic hemostatic assays, which have been used for decades, rely on measuring biomechanical responses to physical stimuli but face challenges related to high device and test cost, limited portability, and limited scalability.. Here, we report a differential pattern using self-induced adaptive-bubble behavioral perception to refresh it. The adaptive behaviors of bubble deformation during coagulation precisely describe the transformation of viscoelastic hemostatic properties, being free of the precise and complex physical devices. And the integrated bubble array chip allows microassays and enables multi-bubble tests with good reproducibility. Recognition of the developed bubble behaviors empowers automated and user-friendly diagnosis. In a prospective clinical study (clinical model development [n = 273]; clinical assay [n = 44]), we show that the diagnostic accuracies were 99.1% for key viscoelastic hemostatic assay indicators (reaction time [R], kinetics time [K], alpha angle [Angle], maximum amplitude [MA], lysis at 30 min [LY30]; n = 220) and 100% (n = 44) for hypercoagulation, healthy, and hypocoagulation diagnoses. This should provide fresh insight into existing paradigms and help more clinical needs.

Digital optofluidic compound eyes with natural structures and zooming capability for large-area fluorescence sensing.

Compound eyes are ubiquitous natural biosensors that possess high temporal resolution and large fields of view (FOVs). While for solid materials based artificial imaging systems, flexible zooming ability while keeping the constant FOV is still challenging, as well as the low-cost fabrication. Herein, liquid compound eyes with natural structures are presented that synthesize optofluidics and bionics in a non-trivial manner, which enables the deformation-free zooming and flexible cell fluorescence sensing. Experimental results indicate that the innovatively manufactured bionic template possesses low roughness and uniform lens configuration with more than two thousands units, which endows the eyes with high-quality and low aberration imaging ability. Besides, digital controlled miscible liquids switching enables the focus of ommatidia simultaneously be adjusted from 150 µm to 5 mm with 100° view angle, and without bending the microlens curvature, to avoid FOV changing and image aberration. Due to large FOV and tunable ability, large-area cell fluorescence signal arrays and dynamic cell motion are imaged using this liquid compound eyes. This work presents novel strategy for compound lens manufacture at low-cost, and proposes deformation-free and continuous focus-tuning strategy, offering potentials for numerous applications, including biomedical sensing and adaptive imaging with large FOV.

Space-time-regulated imaging analyzer for smart coagulation diagnosis.

The development of intelligent blood coagulation diagnoses is awaited to meet the current need for large clinical time-sensitive caseloads due to its efficient and automated diagnoses. Herein, a method is reported and validated to realize it through artificial intelligence (AI)-assisted optical clotting biophysics (OCB) properties identification. The image differential calculation is used for precise acquisition of OCB properties with elimination of initial differences, and the strategy of space-time regulation allows on-demand space time OCB properties identification and enables diverse blood function diagnoses. The integrated applications of smartphones and cloud computing offer a user-friendly automated analysis for accurate and convenient diagnoses. The prospective assays of clinical cases (n = 41) show that the system realizes 97.6%, 95.1%, and 100% accuracy for coagulation factors, fibrinogen function, and comprehensive blood coagulation diagnoses, respectively. This method should enable more low-cost and convenient diagnoses and provide a path for potential diagnostic-markers finding.

A Phosphorescence Quenching-Based Intelligent Dissolved Oxygen Sensor on an Optofluidic Platform.

Continuous measurement of dissolved oxygen (DO) is essential for water quality monitoring and biomedical applications. Here, a phosphorescence quenching-based intelligent dissolved oxygen sensor on an optofluidic platform for continuous measurement of dissolved oxygen is presented. A high sensitivity dissolved oxygen-sensing membrane was prepared by coating the phosphorescence indicator of platinum(II) meso-tetrakis(pentafluorophenyl)porphyrin (PtTFPP) on the surface of the microfluidic channels composed of polydimethylsiloxane (PDMS) microstructure arrays. Then, oxygen could be determined by its quenching effect on the phosphorescence, according to Stern-Volmer model. The intelligent sensor abandons complicated optical or electrical design and uses a photomultiplier (PMT) counter in cooperation with a mobile phone application program to measure phosphorescence intensity, so as to realize continuous, intelligent and real-time dissolved oxygen analysis. Owing to the combination of the microfluidic-based highly sensitive oxygen sensing membrane with a reliable phosphorescent intensity detection module, the intelligent sensor achieves a low limit of detection (LOD) of 0.01 mg/L, a high sensitivity of 16.9 and a short response time (22 s). Different natural water samples were successfully analyzed using the intelligent sensor, and results demonstrated that the sensor features a high accuracy. The sensor combines the oxygen sensing mechanism with optofluidics and electronics, providing a miniaturized and intelligent detection platform for practical oxygen analysis in different application fields.

Rapid nitrate determination with a portable lab-on-chip device based on double microstructured assisted reactors.

Determining the nitrate levels is critical for water quality monitoring, and traditional methods are limited by high toxicity and low detection efficiency. Here, rapid nitrate determination was realized using a portable device based on innovative three-dimensional double microstructured assisted reactors (DMARs). On-chip nitrate reduction and chromogenic reaction were conducted in the DMARs, and the reaction products then flowed into a PMMA optical detection chip for absorbance measurement. A significant enhancement of reaction rate and efficiency was observed in the DMARs due to their sizeable surface-area-to-volume ratios and hydrodynamics in the microchannels. The highest reduction ratio of 94.8% was realized by optimizing experimental parameters, which is greatly improved compared to conventional zinc-cadmium based approaches. Besides, modular optical detection improves the reliability of the portable device, and a smartphone was used to achieve a portable and convenient nitrate analysis. Different water samples were successfully analysed using the portable device based on DMARs. The results demonstrated that the device features fast detection (115 s per sample), low reagent consumptions (26.8 µL per sample), particularly low consumptions of toxic reagents (0.38 µL per sample), good reproducibility and low relative standard deviations (RSDs, 0.5-1.38%). Predictably, the portable lab-on-chip device based on microstructured assisted reactors will find more applications in the field of water quality monitoring in the near future.

On-chip rapid drug screening of leukemia cells by acoustic streaming.

Rapid and personalized single-cell drug screening testing plays an essential role in acute myeloid leukemia drug combination chemotherapy. Conventional chemotherapeutic drug screening is a time-consuming process because of the natural resistance of cell membranes to drugs, and there are still great challenges related to using technologies that change membrane permeability such as sonoporation in high-throughput and precise single-cell drug screening with minimal damage. In this study, we proposed an acoustic streaming-based non-invasive single-cell drug screening acceleration method, using high-frequency acoustic waves (>10 MHz) in a concentration gradient microfluidic device. High-frequency acoustics leads to increased difficulties in inducing cavitation and generates acoustic streaming around each single cell. Therefore, single-cell membrane permeability is non-invasively increased by the acoustic pressure and acoustic streaming-induced shear force, which significantly improves the drug uptake process. In the experiment, single human myeloid leukemia mononuclear (THP-1) cells were trapped by triangle cell traps in concentration gradient chips with different cytarabine (Ara-C) drug concentrations. Due to this dual acoustic effect, the drugs affect cell viability in less than 30 min, which is faster than traditional methods (usually more than 24 h). This dual acoustic effect-based drug delivery strategy has the potential to save time and reduce the cost of drug screening, when combined with microfluidic technology for multi-concentration drug screening. This strategy offers enormous potential for use in multiple drug screening or efficient drug combination screening in individualized/personalized treatments, which can greatly improve efficiency and reduce costs.

Touchable cell biophysics property recognition platforms enable multifunctional blood smart health care.

As a crucial biophysical property, red blood cell (RBC) deformability is pathologically altered in numerous disease states, and biochemical and structural changes occur over time in stored samples of otherwise normal RBCs. However, there is still a gap in applying it further to point-of-care blood devices due to the large external equipment (high-resolution microscope and microfluidic pump), associated operational difficulties, and professional analysis. Herein, we revolutionarily propose a smart optofluidic system to provide a differential diagnosis for blood testing via precise cell biophysics property recognition both mechanically and morphologically. Deformation of the RBC population is caused by pressing the hydrogel via an integrated mechanical transfer device. The biophysical properties of the cell population are obtained by the designed smartphone algorithm. Artificial intelligence-based modeling of cell biophysics properties related to blood diseases and quality was developed for online testing. We currently achieve 100% diagnostic accuracy for five typical clinical blood diseases (90 megaloblastic anemia, 78 myelofibrosis, 84 iron deficiency anemia, 48 thrombotic thrombocytopenic purpura, and 48 thalassemias) via real-world prospective implementation; furthermore, personalized blood quality (for transfusion in cardiac surgery) monitoring is achieved with an accuracy of 96.9%. This work suggests a potential basis for next-generation blood smart health care devices.

Autonomous and In Situ Ocean Environmental Monitoring on Optofluidic Platform.

Determining the distributions and variations of chemical elements in oceans has significant meanings for understanding the biogeochemical cycles, evaluating seawater pollution, and forecasting the occurrence of marine disasters. The primary chemical parameters of ocean monitoring include nutrients, pH, dissolved oxygen (DO), and heavy metals. At present, ocean monitoring mainly relies on laboratory analysis, which is hindered in applications due to its large size, high power consumption, and low representative and time-sensitive detection results. By integrating photonics and microfluidics into one chip, optofluidics brings new opportunities to develop portable microsystems for ocean monitoring. Optofluidic platforms have advantages in respect of size, cost, timeliness, and parallel processing of samples compared with traditional instruments. This review describes the applications of optofluidic platforms on autonomous and in situ ocean environmental monitoring, with an emphasis on their principles, sensing properties, advantages, and disadvantages. Predictably, autonomous and in situ systems based on optofluidic platforms will have important applications in ocean environmental monitoring.

Versatile biomimetic array assembly by phase modulation of coherent acoustic waves.

A high-throughput cell-assembly method, with the advantages of adjustability, ease of operation, and good precision, is remarkable for artificial tissue engineering. Here, we present a scientific solution by introducing high rotational symmetrical coherent acoustic waves, in order to enable the shape and arrangement of the acoustic potential wells to be flexibly modulated, and therefore to assemble on a large area diverse biomimetic arrays on a microfluidic platform. Ring arrays, honeycomb, and many other biomimetic arrays are achieved by real-time modulation of the wave vectors and phase relation of acoustic beams from six directions. In the experiments, human umbilical vein endothelial cells (HUVECs), arranged in ring structures, tend to connect with the adjacent cells and reach confluency, thus directing the in vitro two-dimensional vascular network formation. Higher rotational symmetry of the six coherent acoustic waves provides much more flexibility and diversity for acoustic cell assembly. With the advantages of efficiency, diversity and adjustability, this acoustic chip is expected to fulfill many applications, such as in biochemistry, bioprinting and tissue engineering related research.

A Portable and Accurate Phosphate Sensor Using a Gradient Fabry-Pérot Array.

Here, a portable and accurate phosphate sensor using a gradient Fabry-Pérot array (FPA) is proposed. It can form a bidirectional gradient concentration (absorbance) distribution in the gradient FPA, simplifying the complex operations to get a standard curve and saving time. The gradient FPA can effectively filter out the interference (bubbles, light intensity, and salinity) while improving the absorbance, achieving a highly accurate and stable detection. Besides, the smartphone simplifies data processing and makes sensors more portable. In this work, the detection errors of standard solutions (100, 50, and 30 µM) are 0.39, 1.48, and 1.84%, respectively, and it has also been demonstrated with errors of 2.46 (sample 1, seawater), 2.08 (sample 2, lake water), and 1.83% (sample 3, sewage) for natural samples detection, which is more accurate than a traditional analyzer. The sensor has a good performance when affected by bubbles, light intensity, and salinity. In addition, the detection time is shortened to 80 s, which is more time saving compared with traditional devices, and the limit of detection (LOD) is 0.4 µM. It can be predicted that the novel optofluidic sensor is conducive to build a smart nutrient monitoring system and will be applied in the field of biochemistry and environmental chemistry.

On-chip hydrogel arrays individually encapsulating acoustic formed multicellular aggregates for high throughput drug testing.

Multicellular aggregates in three-dimensional (3D) environments provide novel solid tumor models that can provide insight into in vivo drug resistance. Such models are therefore essential for developing new drugs and preventing the failure of clinical treatments. However, high-throughput cell cluster assembly and fabricating individual 3D environments that mimic the extracellular matrix (ECM) remain significant challenges. To rapidly produce mini 3D multicellular aggregate units, acoustic force assembly combined with ECM mimic hydrogel array encapsulation is developed and then integrated into a diffusion-based microfluidic device for high-throughput drug testing. The active acoustic force gathers human mononuclear leukemia cells (THP-1) into hundreds of multicellular clusters with a controllable size. Instead of continuous bulk materials, photosensitive gelatin methacryloyl (GelMA) hydrogel pillar arrays containing cell clusters at drug concentration gradients are obtained through selective area exposure. Ten azelaic acid (AZA) concentration gradient series are applied to 100 units to simultaneously test the multicellular cluster drug resistance to multiple drug conditions. Real-time green fluorescent protein (GFP) fluorescence is analyzed to monitor cell viability. The results show that cell aggregate activity is inversely related to the drug concentration in the hydrogel pillars, and shows lower sensitivity to drug toxicity than the activity of monolayer cultured cells. The 3D multicellular arrays provide numerous in vitro tumor models and can be directly used for downstream drug testing. This technology inherits the advantages of acoustic assembly, while being more flexible, practical, and high-throughput, and shows significant potential for use in further tumor related research and clinical practice.