Passive Microfluidic Device Research Articles

Parametric study on the margination of white blood cells (WBCs) in a passive microfluidic device

From fundamental research exploring molecular mechanisms to advanced endeavours like the development of artificial white blood cells, white blood cells have widespread applications due to their crucial role in safeguarding the body, defending against infection, and eliminating external pathogens, serving as indicators of several diseases. So, white blood cells must be separated efficiently with high purity to make the passive microdevices more effective for diagnostic applications. In this study, we investigated how cell concentration and viscosity collectively impact the margination of white blood cells in an inertial flow microchannel. We aim to improve white blood cell separation efficiency in microfluidic devices. Unlike previous studies that examined individual factors, this research focused on the interplay between bio-physical properties like cell interaction and viscosity. To explore their correlation, we diluted the blood with phosphate-buffered saline and viscoelastic fluid to manipulate the viscosity and cell concentration. Viscoelastic fluid improved white blood cells margination, while higher viscosity and low cell concentration decreased it. Maintaining blood-like viscosity, optimizing cell-free layer width, and promoting cell-cell interaction in the focusing zone collectively enhanced white blood cells margination.

Optimization of Microfluidics for Point-of-Care Blood Sensing.

Blood tests are widely used in modern medicine to diagnose certain illnesses and evaluate the overall health of a patient. To enable testing in resource-limited areas, there has been increasing interest in point-of-care (PoC) testing devices. To process blood samples, liquid mixing with active pumps is usually required, making PoC blood testing expensive and bulky. We explored the possibility of processing approximately 2 μL of whole blood for image flow cytometry using capillary structures that allowed test times of a few minutes without active pumps. Capillary pump structures with five different pillar shapes were simulated using Ansys Fluent to determine which resulted in the fastest whole blood uptake. The simulation results showed a strong influence of the capillary pump pillar shape on the chip filling time. Long and thin structures with a high aspect ratio exhibited faster filling times. Microfluidic chips using the simulated pump design with the most efficient blood uptake were fabricated with polydimethylsiloxane (PDMS) and polyethylene oxide (PEO). The chip filling times were tested with 2 μL of both water and whole blood, resulting in uptake times of 24 s for water and 111 s for blood. The simulated blood plasma results deviated from the experimental filling times by about 35% without accounting for any cell-induced effects. By comparing the flow speed induced by different pump pillar geometries, this study offers insights for the design and optimization of passive microfluidic devices for inhomogenous liquids such as whole blood in sensing applications.

Cotton threads encapsulated by thermal contraction tube for point-of-care diagnostics

Passive microfluidic devices based on capillary force are popular choices for point-of-care diagnostics. Paper, threads and cloth have been used to fabricate these microfluidic devices. Here, we introduced a novel method for packaging microfluidic thread-based analytical devices (μTADs) by encapsulating cotton threads in thermal contraction tubes. This package method can help to increase convenience during practice. At first, we investigate the influence of this package method on the capillary flow behavior and water absorption capacity of cotton threads. Then, we show the possibility of this package method in preparing μTADs with complex shapes. Lastly, we successfully use μTADs based on this package method for applications in point-of-care diagnostics including glucose detection in urine, fibrinogen detection in blood plasma, and plasma separation from whole blood samples and hematocrit (HCT) measurement. The package method is promising in expanding the applications of μTADs in point-of-care diagnostics.

Numerical study of bacteria removal from microalgae solution using an asymmetric contraction-expansion microfluidic device: A parametric analysis approach

Microalgae have been remarkably taken into account due to their wide applications in the biopharmaceutical, nutraceutical and bio-energy fields. However, contamination of microalgae with bacteria still appears to be a concern, adversely impacting products’ quality and process efficiency. Microalgae decontamination with conventional techniques is usually expensive and time-consuming. Moreover, damage to microalgae cells is highly possible. Asymmetric contraction-expansion microchannels (Asym-CEMCs) are promising passive microfluidic devices that can overcome conventional techniques' drawbacks with their standing-out features. However, the flexibility of Asym-CEMCs performance arising from their various tunable geometrical parameters results in the fact that their performance for separating a target particle cannot be predicted without an investigation. In this work, for the first time, Asym-CEMCs were numerically studied for the removal of a very conventional bacteria, B. subtilis (1 μm), from one of the most popular microalgae, C. vulgaris (5.7 μm). The influences of the microchannel aspect ratio, length and width ratios of the expansion-to-contraction zones, and the total flow rate on the separation resolution and focusing width of the particles were investigated by a 3D numerical model. The aspect ratio had the strongest influence on the Asym-CEMC performance, however, the length ratio had no considerable effect on the results. A decrease in the aspect ratio augmented the shear-induced lift force and Dean drag force, leading to a significant separation resolution improvement. Microalgae decontamination was also enhanced by an increase in the total flow rate and expansion-to-contraction width ratio. Finally, a locally optimized Asym-CEMC with an aspect ratio of one and expansion-to-contraction width and length ratios of 4.7 and 2.07, respectively, was proposed, leading to complete microalgae decontamination with a high normalized separation resolution of 0.6. In a word, Asym-CEMCs with tailored dimensions are promising for successfully decontaminating microalgae from bacteria.

Blood component separation in straight microfluidic channels.

Separation of blood components is required in many diagnostic applications and blood processes. In laboratories, blood is usually fractionated by manual operation involving a bulk centrifugation equipment, which significantly increases logistic burden. Blood sample processing in the field and resource-limited settings cannot be readily implemented without the use of microfluidic technology. In this study, we developed a small footprint, rapid, and passive microfluidic channel device that relied on margination and inertial focusing effects for blood component separation. No blood dilution, lysis, or labeling step was needed as to preserve sample integrity. One main innovation of this work was the insertion of fluidic restrictors at outlet ports to divert the separation interface into designated outlet channels. Thus, separation efficiency was significantly improved in comparison to previous works. We demonstrated different operation modes ranging from platelet or plasma extraction from human whole blood to platelet concentration from platelet-rich plasma through the manipulation of outlet port fluidic resistance. Using straight microfluidic channels with a high aspect ratio rectangular cross section, we demonstrated 95.4% platelet purity extracted from human whole blood. In plasma extraction, 99.9% RBC removal rate was achieved. We also demonstrated 2.6× concentration of platelet-rich plasma solution to produce platelet concentrate. The extraction efficiency and throughput rate are scalable with continuous and clog-free recirculation operation, in contrast to other blood fractionation approaches using filtration membranes or affinity-based purification methods. Our microfluidic blood separation method is highly tunable and versatile, and easy to be integrated into multi-step blood processing and advanced sample preparation workflows.

A layout-dependent formula for the critical separation size of parallelogram-type deterministic lateral displacement arrays

The accurate and general formulation of the critical separation size is crucial for the effective design and application of passive microfluidic devices. However, the current formulas for the deterministic lateral displacement (DLD) technique are inadequate in accounting for its structural complexity. To address this limitation, we conducted mesoscopic hydrodynamics simulations to assess the separation performance of various parallelogram-type circular post arrays. Based on the simulation results, we developed a new layout-dependent formula that takes into account key geometric parameters, such as the lateral gap size, row shift fraction, and aspect ratio of downstream and lateral post-post distances, to characterize the lateral gap dimension, periodicity, and asymmetry of DLD arrays, respectively. This formula demonstrated high accuracy over a wide design space, precisely predicting the critical separation sizes of many asymmetric DLD devices in experiments. Furthermore, it is worth noting that when the downstream gap size expands, the veering flow strengthens, which, in turn, results in an increase in the critical separation size. These findings shed light on the unique separation mechanism induced by array asymmetry and present a powerful design tool for maximizing the potential of asymmetric DLD devices.

Red blood cell rosetting enables size-based separation of specific lymphocyte subsets from blood in a microfluidic device.

The isolation of a specific lymphocyte subset from blood is the required first step in the manufacturing of many novel cellular immunotherapies. Microfluidic size-based separation methods are poised to significantly simplify this process because they require neither centrifugation nor magnetic or fluorescent labeling to operate. Lymphocytes can be separated from red blood cells (RBCs) and platelets as well as monocytes and granulocytes because their size differs from each of these cell types. However, further separation of a specific lymphocyte subset from other unwanted lymphocytes using size-based methods is impossible because all lymphocytes have approximately the same size and can only be distinguished by surface markers. This paper describes a new approach that made it possible for a size-based separation method to isolate a desired subset of lymphocytes by making unwanted lymphocytes as well as other blood cells artificially larger. The separation was enabled by selectively binding multiple RBCs to each unwanted cell to create 'rosettes' with an effective size significantly larger than the diameter of a typical lymphocyte. The desired lymphocytes remained unaffected by rosetting and were separated from the rosettes by passing the mixture through a microfluidic size-based separation device based on controlled incremental filtration (CIF). This new rosette-enabled size-based (RESIZE) separation approach demonstrated recovery of 80-90% for all lymphocyte subsets tested (CD3+, CD4+, CD56+) which was ∼2.5-fold higher than that for the standard immunodensity method (RBC rosetting followed by density gradient centrifugation). The purity of separation was >90% for CD3+ cells but declined with increasing cell rarity. Unlike the immunodensity approach, RESIZE required neither centrifugation nor cell washing after the separation and was ∼2.5-fold faster when processing the same sample volume. The results of this study suggest that integration of the RESIZE approach for high-yield isolation of lymphocyte subsets from blood could significantly streamline the manufacturing workflow and thus have a potentially transformative impact on the cost and availability of novel cellular immunotherapies.

Biomedical Applications of Microfluidic Devices: A Review.

Both passive and active microfluidic chips are used in many biomedical and chemical applications to support fluid mixing, particle manipulations, and signal detection. Passive microfluidic devices are geometry-dependent, and their uses are rather limited. Active microfluidic devices include sensors or detectors that transduce chemical, biological, and physical changes into electrical or optical signals. Also, they are transduction devices that detect biological and chemical changes in biomedical applications, and they are highly versatile microfluidic tools for disease diagnosis and organ modeling. This review provides a comprehensive overview of the significant advances that have been made in the development of microfluidics devices. We will discuss the function of microfluidic devices as micromixers or as sorters of cells and substances (e.g., microfiltration, flow or displacement, and trapping). Microfluidic devices are fabricated using a range of techniques, including molding, etching, three-dimensional printing, and nanofabrication. Their broad utility lies in the detection of diagnostic biomarkers and organ-on-chip approaches that permit disease modeling in cancer, as well as uses in neurological, cardiovascular, hepatic, and pulmonary diseases. Biosensor applications allow for point-of-care testing, using assays based on enzymes, nanozymes, antibodies, or nucleic acids (DNA or RNA). An anticipated development in the field includes the optimization of techniques for the fabrication of microfluidic devices using biocompatible materials. These developments will increase biomedical versatility, reduce diagnostic costs, and accelerate diagnosis time of microfluidics technology.

Label-free deep-UV microscopy detection and grading of neutropenia using a passive microfluidic device.

Neutropenia is a condition comprising an abnormally low number of neutrophils, a type of white blood cell, which puts patients at an increased risk of severe infections. Neutropenia is especially common among cancer patients and can disrupt their treatment or even be life-threatening in severe cases. Therefore, routine monitoring of neutrophil counts is crucial. However, the current standard of care to assess neutropenia, the complete blood count (CBC), is resource-intensive, time-consuming, and expensive, thereby limiting easy or timely access to critical hematological information such as neutrophil counts. Here, we present a simple technique for fast, label-free neutropenia detection and grading via deep-ultraviolet (deep-UV) microscopy of blood cells in polydimethylsiloxane (PDMS)-based passive microfluidic devices. The devices can potentially be manufactured in large quantities at a low cost, requiring only 1 μL of whole blood for operation. We show that the absolute neutrophil counts (ANC) obtained from our proposed microfluidic device-enabled deep-UV microscopy system are highly correlated with those from CBCs using commercial hematology analyzers in patients with moderate and severe neutropenia, as well as healthy donors. This work lays the foundation for the development of a compact, easy-to-use UV microscope system to track neutrophil counts that is suitable for low-resource, at-home, or point-of-care settings.

Microfluidic Sampling and Biosensing Systems for Foodborne Escherichia coli and Salmonella.

Developments of portable biosensors for field-deployable detections have been increasingly important to control foodborne pathogens in regulatory environment and in early stage of outbreaks. Conventional cultivation and gene amplification methods require sophisticated instruments and highly skilled professionals; while portable biosensing devices provide more freedom for rapid detections not only in research laboratories but also in the field; however, their sensitivity and specificity are limited. Microfluidic methods have the advantage of miniaturizing instrumental size while integrating multiple functions and high-throughput capability into one streamlined system at low cost. Minimal sample consumption is another advantage to detect samples in different sizes and concentrations, which is important for the close monitoring of pathogens at consumer end. They improve measurement or manipulation of bacteria by increasing the ratio of functional interface of the device to the targeted biospecies and in turn reducing background interference. This article introduces the major active and passive microfluidic devices that have been used for bacteria sampling and biosensing. The emphasis is on particle-based sorting/enrichment methods with or without external physical fields applied to the microfluidic devices and on various biosensing applications reported for bacteria sampling. Three major fabrication methods for microfluidics are briefly discussed with their advantages and limitations. The applications of these active and passive microfluidic sampling methods in the past 5 years have been summarized, with the focus on Escherichia coli and Salmonella. The current challenges to microfluidic bacteria sampling are caused by the small size and nonspherical shape of various bacterial cells, which can induce unpredictable deviations in sampling and biosensing processes. Future studies are needed to develop rapid prototyping methods for device manufacturing, which can facilitate rapid response to various foodborne pathogen outbreaks.

Cover Feature: Disposable Passive Electrochemical Microfluidic Device for Diagnosis of Congenital Disorders of Glycosylation (Anal. Sens. 1/2022)

The cover feature shows disposable passive electrochemical microfluidic devices which were developed using a lamination technique for the diagnosis of congenital glycosylation disorders. The key assay steps were integrated on board with high reproducibility. It was possible to successfully discriminate between patients and healthy individuals by taking short analysis times with minimal analyst intervention and requiring low clinical sample volumes. These results suggest that these devices have potential as future tools at the point of patient care. The cover was drawn by Víctor de la Asunción and co-designed with the authors. More information can be found in the Full Paper by Alberto Escarpa, Agustín G. Crevillén and co-workers.

Disposable Passive Electrochemical Microfluidic Device for Diagnosis of Congenital Disorders of Glycosylation

AbstractA disposable pump‐free electrochemical microfluidic device made by using a multilayer lamination technique is proposed for diagnosis of glycosylation disorders. Thanks to the stacking of polyethylene terephthalate (PET) films and double adhesive layers, it is possible to create a geometry that allows the filling of the tailored channels in a fixed time. In the main channel of this passive microfluidic device, the necessary steps to perform the electrochemical determination of transferrin are performed: labeling of the protein with Os (VI) complex (electrochemical tag), washing with water of the osmium residues and electrochemical detection by adsorptive transfer stripping square wave voltammetry. Electrochemical detection released two electrochemical signals: one from Os (VI) complex due to carbohydrates at −0.9 V, and other from the intrinsic electrochemical signal of glycoprotein due to the amino acids at +0.8 V. The ratio between them establishes an indicator of the degree of glycosylation (called electrochemical index of glycosylation). The method was successfully applied to the analysis of clinical samples from patients with congenital disorders of glycosylation.

High-Performance Passive Plasma Separation on OSTE Pillar Forest.

Plasma separation is of high interest for lateral flow tests using whole blood as sample liquids. Here, we built a passive microfluidic device for plasma separation with high performance. This device was made by blood filtration membrane and off-stoichiometry thiol–ene (OSTE) pillar forest. OSTE pillar forest was fabricated by double replica moldings of a laser-cut polymethylmethacrylate (PMMA) mold, which has a uniform microstructure. This device utilized a filtration membrane to separate plasma from whole blood samples and used hydrophilic OSTE pillar forest as the capillary pump to propel the plasma. The device can be used to separate blood plasma with high purity for later use in lateral flow tests. The device can process 45 L of whole blood in 72 s and achieves a plasma separation yield as high as 60.0%. The protein recovery rate of separated plasma is 85.5%, which is on par with state-of-the-art technologies. This device can be further developed into lateral flow tests for biomarker detection in whole blood.

Embedded screen-printed transducers in bulk polymer microfluidic devices

We present a low-cost method to fabricate microfluidic devices with embedded transducers suitable for rapid prototyping on a lab scale and potentially also for mass production. To achieve this goal, a method for fabricating passive microfluidic devices using a solvent bonding technique for sealing is introduced. To avoid clogging, liquid metal (gallium) is utilized as a sacrificial layer during this bonding process. A sensor fabrication method compatible with the rapid prototyping ambitions is screen printing. Sensors based on thermal, capacitive and mechanical effects are fabricated by screen printing and are embedded into channels. The proposed processing methods can be easily adapted to achieve specific requirements associated with different tasks in lab-on-a-chip devices.

A Disposable Passive Microfluidic Device for Cell Culturing.

In this work, a disposable passive microfluidic device for cell culturing that does not require any additional/external pressure sources is introduced. By regulating the height of fluidic columns and the aperture and closure of the source wells, the device can provide different media and/or drug flows, thereby allowing different flow patterns with respect to time. The device is made of two Polymethylmethacrylate (PMMA) layers fabricated by micro-milling and solvent assisted bonding and allows us to ensure a flow rate of 18.6 μL/h - 7%/day, due to a decrease of the fluid height while the liquid is driven from the reservoirs into the channels. Simulations and experiments were conducted to characterize flows and diffusion in the culture chamber. Melanoma tumor cells were used to test the device and carry out cell culturing experiments for 48 h. Moreover, HeLa, Jurkat, A549 and HEK293T cell lines were cultivated successfully inside the microfluidic device for 72 h.

Directed Placement for mVLSI Devices

Continuous-flow microfluidic devices based on integrated channel networks are becoming increasingly prevalent in research in the biological sciences. At present, these devices are physically laid out by hand by domain experts who understand both the underlying technology and the biological functions that will execute on fabricated devices. The lack of a design science that is specific to microfluidic technology creates a substantial barrier to entry. To address this concern, this article introduces Directed Placement, a physical design algorithm that leverages the natural “directedness” in most modern microfluidic designs: fluid enters at designated inputs, flows through a linear or tree-based network of channels and fluidic components, and exits the device at dedicated outputs. Directed placement creates physical layouts that share many principle similarities to those created by domain experts. Directed placement allows components to be placed closer to their neighbors compared to existing layout algorithms based on planar graph embedding or simulated annealing, leading to an average reduction in laid-out fluid channel length of 91% while improving area utilization by 8% on average. Directed placement is compatible with both passive and active microfluidic devices and is compatible with a variety of mainstream manufacturing technologies.

A Microfluidic Deformability Assessment of Pathological Red Blood Cells Flowing in a Hyperbolic Converging Microchannel.

The loss of the red blood cells (RBCs) deformability is related with many human diseases, such as malaria, hereditary spherocytosis, sickle cell disease, or renal diseases. Hence, during the last years, a variety of technologies have been proposed to gain insights into the factors affecting the RBCs deformability and their possible direct association with several blood pathologies. In this work, we present a simple microfluidic tool that provides the assessment of motions and deformations of RBCs of end-stage kidney disease (ESKD) patients, under a well-controlled microenvironment. All of the flow studies were performed within a hyperbolic converging microchannels where single-cell deformability was assessed under a controlled homogeneous extensional flow field. By using a passive microfluidic device, RBCs passing through a hyperbolic-shaped contraction were measured by a high-speed video microscopy system, and the velocities and deformability ratios (DR) calculated. Blood samples from 27 individuals, including seven healthy controls and 20 having ESKD with or without diabetes, were analysed. The obtained data indicates that the proposed device is able to detect changes in DR of the RBCs, allowing for distinguishing the samples from the healthy controls and the patients. Overall, the deformability of ESKD patients with and without diabetes type II is lower in comparison with the RBCs from the healthy controls, with this difference being more evident for the group of ESKD patients with diabetes. RBCs from ESKD patients without diabetes elongate on average 8% less, within the hyperbolic contraction, as compared to healthy controls; whereas, RBCs from ESKD patients with diabetes elongate on average 14% less than the healthy controls. The proposed strategy can be easily transformed into a simple and inexpensive diagnostic microfluidic system to assess blood cells deformability due to the huge progress in image processing and high-speed microvisualization technology.

Blood Cells Separation and Sorting Techniques of Passive Microfluidic Devices: From Fabrication to Applications.

Since the first microfluidic device was developed more than three decades ago, microfluidics is seen as a technology that exhibits unique features to provide a significant change in the way that modern biology is performed. Blood and blood cells are recognized as important biomarkers of many diseases. Taken advantage of microfluidics assets, changes on blood cell physicochemical properties can be used for fast and accurate clinical diagnosis. In this review, an overview of the microfabrication techniques is given, especially for biomedical applications, as well as a synopsis of some design considerations regarding microfluidic devices. The blood cells separation and sorting techniques were also reviewed, highlighting the main achievements and breakthroughs in the last decades.

A Passive Microfluidic Device for Chemotaxis Studies.

This work presents a disposable passive microfluidic system, allowing chemotaxis studies, through the generation of a concentration gradient. The device can handle liquid flows without an external supply of pressure or electric gradients, but simply using gravity force. It is able to ensure flow rates of 10 µL/h decreasing linearly with 2.5% in 24 h. The device is made of poly(methylmethacrylate) (PMMA), a biocompatible material, and it is fabricated by micro-milling and solvent assisted bonding. It is assembled into a mini incubator, designed properly for cell biology studies in passive microfluidic devices, which provides control of temperature and humidity levels, a contamination-free environment for cells with air and 5% of CO2. Furthermore, the mini incubator can be mounted on standard inverted optical microscopes. By using our microfluidic device integrated into the mini incubator, we are able to evaluate and follow in real-time the migration of any cell line to a chemotactic agent. The device is validated by showing cell migration at a rate of 0.36 µm/min, comparable with the rates present in scientific literature.

Fully Stretchable Capillary Microfluidics-Integrated Nanoporous Gold Electrochemical Sensor for Wearable Continuous Glucose Monitoring.

Biosensor systems for wearable continuous monitoring are desired to be developed into conformal patch platforms. However, developing such patches is very challenging owing to the difficulty of imparting materials and components with both high stretchability and high performance. Herein, we report a fully stretchable microfluidics-integrated glucose sensor patch comprised of an omnidirectionally stretchable nanoporous gold (NPG) electrochemical biosensor and a stretchable passive microfluidic device. A highly electrocatalytic NPG electrode was formed on a stress-absorbing 3D micropatterned polydimethylsiloxane (PDMS) substrate to confer mechanical stretchability, high sensitivity, and durability in non-enzymatic glucose detection. A thin, stretchable, and tough microfluidic device was made by embedding stretchable cotton fabric as a capillary into a thin polyurethane nanofiber-reinforced PDMS channel, enabling collection and passive, accurate delivery of sweat from skin to the electrode surface, with excellent replacement capability. The integrated glucose sensor patch demonstrated excellent ability to continuously and accurately monitor the sweat glucose level.