Microfluidic Tools Research Articles

Microfluidics-A novel technique for high-quality sperm selection for greater ART outcomes.

Microfluidics represent a quality sperm selection technique. Human couples fail to conceive and this is so in a significant population of animals worldwide. Defects in male counterpart lead to failure of conception so are outcomes of assisted reproduction affected by quality of sperm. Microfluidics, deals with minute volumes (μL) of liquids run in small-scale microchannel networks in the form of laminar flow streamlines. Microfluidic sperm selection designs have been developed in chip formats, mimicking invivo situations. Here sperms are selected and analyzed based on motility and sperm behavioral properties. Compared to conventional sperm selection methods, this selection method enables to produce high-quality motile sperm cells possessing non-damaged or least damaged DNA, achieve greater success of insemination in bovines, and achieve enhanced pregnancy rates and live births in assisted reproduction-in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Besides, the concentration of sperm available to oocyte can be controlled by regulating the flow rate in microfluidic chips. The challenges in this technology are commercialization of chips, development of fully functional species-specific microfluidic tools, limited number of studies available in literature, and need of thorough understanding in reproductive physiology of domestic animals. In conclusion, incorporation of microfluidic system in assisted reproduction for sperm selection may promise a great success in IVF and ICSI outcomes. Future prospectives are to make this technology more superior and need to modify chip designs which is cost effective and species specific and ready for commercialization. Comprehensive studies in animal species are needed to be carried out for wider application of microfluidic sperm selection in invitro procedures.

DROPLET MOTION ON HYDROPHOBIC AND HYDROPHILIC SURFACES USING LATTICE BOLTZMANN METHOD WITH BLOOD TEST MICROPAD APPLICATION

Laboratory sciences and disease diagnostics need accelerated medical tests, ease of testing for patients, and lower diagnostic costs. In this respect, one of the most innovative methods is paper-based microfluidic analysis tools, called micro-PAD. Since the chemical-based micro-PADs need sufficient blood to show an accurate blood test, the shape of the micro-PADs, the blood quantity, and the droplet flow to reach the end of the microchannels are important. Therefore, this research showed the liquid droplet motion on a heterogeneous (hydrophobic and hydrophilic) surface using the lattice Boltzmann method, and two star-shaped micro-PADs were modeled to investigate the effect of the droplet size and the droplet deflection from the center of the micro-PAD on liquid flow in microchannels. Based on the obtained results, fluid behavior was similar in two patterns (ten-pointed pattern and four-pointed pattern). This study compared the effects of parallel, convergent, and divergent channels in a four-pointed pattern on fluid velocity. The off-center error of the droplet was considered at the initial moment. The obtained results confirmed the direct relationship between the increased deviation of the center and the lack of uniformity in the fluid penetration. The present simulation results were validated against the laboratory data.

Molecular fingerprinting of biological nanoparticles with a label-free optofluidic platform

Label-free detection of multiple analytes in a high-throughput fashion has been one of the long-sought goals in biosensing applications. Yet, for all-optical approaches, interfacing state-of-the-art label-free techniques with microfluidics tools that can process small volumes of sample with high throughput, and with surface chemistry that grants analyte specificity, poses a critical challenge to date. Here, we introduce an optofluidic platform that brings together state-of-the-art digital holography with PDMS microfluidics by using supported lipid bilayers as a surface chemistry building block to integrate both technologies. Specifically, this platform fingerprints heterogeneous biological nanoparticle populations via a multiplexed label-free immunoaffinity assay with single particle sensitivity. First, we characterise the robustness and performance of the platform, and then apply it to profile four distinct ovarian cell-derived extracellular vesicle populations over a panel of surface protein biomarkers, thus developing a unique biomarker fingerprint for each cell line. We foresee that our approach will find many applications where routine and multiplexed characterisation of biological nanoparticles are required.

Reducing Cathodic Drift during Isoelectric Focusing Using Microscale Immobilized pH Gradient Gels.

Microfluidic analytical tools play an important role in miniaturizing targeted proteomic assays for improved detection sensitivity, throughput, and automation. Microfluidic isoelectric focusing (IEF) can resolve proteoforms in lysate from low-to-single cell numbers. However, IEF assays often use carrier ampholytes (CAs) to establish a pH gradient for protein separation, presenting limitations like pH instability in the form of cathodic drift (migration of focused proteins toward the cathode). Immobilized pH gradient (IPG) gels reduce cathodic drift by covalently immobilizing the pH buffering components to a matrix. To our knowledge, efforts to implement IPG gels at the microscale have been limited to glass microdevices. To adapt IEF using IPGs to widely used microfluidic device materials, we introduce a polydimethylsiloxane (PDMS)-based microfluidic device and compare the microscale pH gradient stability of IEF established with IPGs, CAs, and a hybrid formulation of IPG gels and CAs (mixed-bed IEF). The PDMS-based IPG microfluidic device (μIPG) resolved analytes differing by 0.1 isoelectric point within a 3.5 mm separation lane over a 20 min focusing duration. During the 20 min duration, we observed markedly different cathodic drift velocities among the three formulations: 60.1 μm/min in CA-IEF, 2.5 μm/min in IPG-IEF (∼24-fold reduction versus CA-IEF), and 1.4 μm/min in mixed-bed IEF (∼43-fold reduction versus CA-IEF). Lastly, mixed-bed IEF in a PDMS device resolved green fluorescent protein (GFP) proteoforms from GFP-expressing human breast cancer cell lysate, thus establishing stability in lysate from complex biospecimens. μIPG is a promising and stable technique for studying proteoforms from small volumes.

Density-contrast induced inertial forces on particles in oscillatory flows

Oscillatory flows have become an indispensable tool in microfluidics, inducing inertial effects for displacing and manipulating fluid-borne objects in a reliable, controllable and label-free fashion. However, the quantitative description of such effects has been confined to limit cases and specialized scenarios. Here we develop an analytical formalism yielding the equation of motion of density-mismatched spherical particles in oscillatory background flows, generalizing previous work. Inertial force terms are systematically derived from the geometry of the flow field together with analytically known Stokes number dependences. Supported by independent, first-principles direct numerical simulations, we find that these forces are important even for nearly density-matched objects such as cells or bacteria, enabling their fast displacement and separation. Our formalism thus consistently incorporates particle inertia into the Maxey–Riley equation, and in doing so provides a generalization of Auton's modification to added mass, as well as recovering the description of acoustic radiation forces on particles as a limiting case.

Two‐Photon Direct Laser Writing of pNIPAM Actuators in Microchannels for Dynamic Microfluidics

Microfluidic tools enable to investigate and manipulate various chemical and biological processes at small scales. As a result, it finds widespread applications in lab‐on‐chip devices, drug delivery systems, or miniaturized cell cultures. However, microfluidic devices are still limited in their flexibility and are often designed to fulfill a single functionality. Moreover, technologies to introduce dynamic functionalities with high precision and at high resolution after the development of a continuous phase microfluidic chip remain scarce. Herein, two‐photon polymerization direct laser writing is introduced as a suitable approach to equip continuous phase microfluidic chips with structurally defined thermoresponsive poly(N‐isopropyl‐acrylamide) (pNIPAM) microactuators. Harnessing the lower critical phase transition temperature of pNIPAM, and upon controlling specific design parameters, the efficient catch and release of polystyrene beads of different sizes using a pNIPAM micropillar brush array is demonstrated. Moreover, a biocompatible pNIPAM microgripper array is designed to subsequently capture and release differently sized (single) cell populations. Overall, the method offers great flexibility and a high degree of freedom toward the fabrication of dynamic microfluidic devices with great adaptability to experimental conditions in real time.

Spiral Large-Dimension Microfluidic Channel for Flow-Rate- and Particle-Size-Insensitive Focusing by the Stabilization and Acceleration of Secondary Flow.

Inertial microfluidics has demonstrated its ability to focus particles in a passive and straightforward manner. However, achieving flow-rate- and particle-size-insensitive focusing in large-dimension channels with a simple design remains challenging. In this study, we developed a spiral microfluidic with a large-dimension channel to achieve inertial focusing. By designing a unique "big buffering area" and a "small buffering area" in the spiral microchannel, we observed the stabilization and acceleration of secondary flow. Our optimized design allowed for efficient (>99.9%) focusing of 15 μm particles within a wide range of flow rates (0.5-4.5 mL/min) during a long operation duration (0-60 min). Additionally, we achieved effective (>95%) focusing of different-sized particles (7, 10, 15, and 30 μm) and three types of tumor cells (K562, HeLa, and MCF-7) near the inner wall of the 1 mm wide outlet when applying different flow rates (1-3 mL/min). Finally, successful 3D cell focusing was achieved within an optimized device, with the cells positioned at a distance of 50 μm from the wall. Our strategy of stabilizing and accelerating Dean-like secondary flow through the unique configuration of a "big buffering area" and a "small buffering area" proved to be highly effective in achieving inertial focusing that is insensitive to the flow rate and particle size, particularly in large-dimension channels. Consequently, it shows great potential for use in hand-operated microfluidic tools for flow cytometry.

Microfluidic fiber spinning for 3D bioprinting: Harnessing microchannels to build macrotissues

Microfluidics is rapidly revolutionizing the scientific panorama, providing unmatched high-throughput platforms that find application in numerous areas of physics, chemistry, biology, and materials science. Recently, microfluidic chips have been proposed, in combination with bioactive materials, as promising tools for spinning cell-laden fibers with on-demand characteristics. However, cells encapsulated in filaments produced via microfluidic spinning technology are confined in a quasi-three-dimensional (3D) environment that fails to replicate the intricate 3D architecture of biological tissues. Thanks to the recent synergistic combination of microfluidic devices with 3D bioprinting technologies that enable the production of sophisticated microfibers serving as the backbone of 3D structures, a new age of tissue engineering is emerging. This review looks at how combining microfluidics with 3D printing is contributing to the biofabrication of relevant human substitutes and implants. This paper also describes the whole manufacturing process from the production of the microfluidic tool to the printing of tissue models, focusing on cutting-edge fabrication technologies and emphasizing the most noticeable achievements for microfluidic spinning technology. A theoretical insight for thixotropic hydrogels is also proposed to predict the fiber size and shear stress developing within microfluidic channels. The potential of using microfluidic chips as bio-printheads for multi-material and multi-cellular bioprinting is discussed, highlighting the challenges that microfluidic bioprinting still faces in advancing the field of biofabrication for tissue engineering and regenerative medicine purposes.

Light-controllable liquid crystal platform for microparticle oscillations and transport.

Liquid crystal colloids manifest complex motion caused by external stimuli, but tunable and addressable control of microsized objects remains a challenge. This study aims to demonstrate light-driven trapping, transport, and sustained periodic motions of microparticles by employing liquid crystal films as a light-controllable colloidal platform. The diverse motions of microscopic particles result from Marangoni convection coupled with elastic deformations in free-surface liquid crystal films subjected to light beam heating. The specific mode of particle motion, including damped and sustained oscillations, also combined with sustained rotation, is defined by the liquid crystal chirality, particle surface treatment, film thickness, and the power of the tightly focused light beam. The results reveal that free-surface liquid crystals provide a unique platform for the indirect optical manipulation of microscopic objects, paving the way for novel applications in microfluidic tools, particle sorting and transport, micropatterning, and various micromachines.

A novel microfluidic tool for the evaluation of local drug delivery systems in simulated in vivo conditions.

A 3D-printed microfluidic tool for assessing local drug delivery systems (LDD) in simulated in vivo conditions was developed and evaluated. The device was designed considering the oral environment and dental applications, and it was fabricated with a high-precision resin 3D printer. Chitosan scaffolds loaded with different concentrations of doxycycline were used for evaluating our device. The concentration of the released drug was measured through in-line UV-VIS spectroscopy, and to verify the repeatability and accuracy of our measurements, comparisons with standard HPLC results were made (5% deviation). Cumulative drug release profiles in steady-state conditions were obtained and compared to the Weibull model. The behaviour of the LDD system in a dynamic environment was also evaluated during experiments where step changes in pH were introduced. It was demonstrated that under infection-like conditions, there is an immediate response from the polymer and a clear increase in the concentration of the released drug. Continuous flow and recirculation experiments were also conducted, revealing significant differences in the drug release profiles. Specifically, in the case of continuous flow, the quantity of the released drug is much higher due to the higher driving force for diffusion (concentration gradient). Overall, the proposed microfluidic tool proved to be ideal for evaluating LDD systems, as the in vivo microenvironment can be replicated in a better way than with currently used standard systems.

Twenty years of islet-on-a-chip: microfluidic tools for dissecting islet metabolism and function.

Pancreatic islets are metabolically active micron-sized tissues responsible for controlling blood glucose through the secretion of insulin and glucagon. A loss of functional islet mass results in type 1 and 2 diabetes. Islet-on-a-chip devices are powerful microfluidic tools used to trap and study living ex vivo human and murine pancreatic islets and potentially stem cell-derived islet organoids. Devices developed over the past twenty years offer the ability to treat islets with controlled and dynamic microenvironments to mimic in vivo conditions and facilitate diabetes research. In this review, we explore the various islet-on-a-chip devices used to immobilize islets, regulate the microenvironment, and dynamically detect islet metabolism and insulin secretion. We first describe and assess the various methods used to immobilize islets including chambers, dam-walls, and hydrodynamic traps. We subsequently describe the surrounding methods used to create glucose gradients, enhance the reaggregation of dispersed islets, and control the microenvironment of stem cell-derived islet organoids. We focus on the various methods used to measure insulin secretion including capillary electrophoresis, droplet microfluidics, off-chip ELISAs, and on-chip fluorescence anisotropy immunoassays. Additionally, we delve into the various multiparametric readouts (NAD(P)H, Ca2+-activity, and O2-consumption rate) achieved primarily by adopting a microscopy-compatible optical window into the devices. By critical assessment of these advancements, we aim to inspire the development of new devices by the microfluidics community and accelerate the adoption of islet-on-a-chip devices by the wider diabetes research and clinical communities.

Microfluidic systems for infectious disease diagnostics.

Microorganisms, encompassing both uni- and multicellular entities, exhibit remarkable diversity as omnipresent life forms in nature. They play a pivotal role by supplying essential components for sustaining biological processes across diverse ecosystems, including higher host organisms. The complex interactions within the human gut microbiota are crucial for metabolic functions, immune responses, and biochemical signalling, particularly through the gut-brain axis. Viruses also play important roles in biological processes, for example by increasing genetic diversity through horizontal gene transfer when replicating inside living cells. On the other hand, infection of the human body by microbiological agents may lead to severe physiological disorders and diseases. Infectious diseases pose a significant burden on global healthcare systems, characterized by substantial variations in the epidemiological landscape. Fast spreading antibiotic resistance or uncontrolled outbreaks of communicable diseases are major challenges at present. Furthermore, delivering field-proven point-of-care diagnostic tools to the most severely affected populations in low-resource settings is particularly important and challenging. New paradigms and technological approaches enabling rapid and informed disease management need to be implemented. In this respect, infectious disease diagnostics taking advantage of microfluidic systems combined with integrated biosensor-based pathogen detection offers a host of innovative and promising solutions. In this review, we aim to outline recent activities and progress in the development of microfluidic diagnostic tools. Our literature research mainly covers the last 5 years. We will follow a classification scheme based on the human body systems primarily involved at the clinical level or on specific pathogen transmission modes. Important diseases, such as tuberculosis and malaria, will be addressed more extensively.

Engineering advancements in microfluidic systems for enhanced mixing at low Reynolds numbers.

Mixing within micro- and millichannels is a pivotal element across various applications, ranging from chemical synthesis to biomedical diagnostics and environmental monitoring. The inherent low Reynolds number flow in these channels often results in a parabolic velocity profile, leading to a broad residence time distribution. Achieving efficient mixing at such small scales presents unique challenges and opportunities. This review encompasses various techniques and strategies to evaluate and enhance mixing efficiency in these confined environments. It explores the significance of mixing in micro- and millichannels, highlighting its relevance for enhanced reaction kinetics, homogeneity in mixed fluids, and analytical accuracy. We discuss various mixing methodologies that have been employed to get a narrower residence time distribution. The role of channel geometry, flow conditions, and mixing mechanisms in influencing the mixing performance are also discussed. Various emerging technologies and advancements in microfluidic devices and tools specifically designed to enhance mixing efficiency are highlighted. We emphasize the potential applications of micro- and millichannels in fields of nanoparticle synthesis, which can be utilized for biological applications. Additionally, the prospects of machine learning and artificial intelligence are offered toward incorporating better mixing to achieve precise control over nanoparticle synthesis, ultimately enhancing the potential for applications in these miniature fluidic systems.

Microfluidic Localized Hydrogel Polymerization Enables Simultaneous Recording of Neural Activity and Behavior in C. elegans.

Monitoring an animal's brain activity during motion provides a means to interpret the brain activity in the context of movement. However, it is challenging to obtain information about the animal's movement during neural imaging in the popular model organism C. elegans due to its small size. Here, we present a microfluidic tool to immobilize only the head region of C. elegans for simultaneous recording of neuronal activity and tail movement. We combine hydrogel photopolymerization and microfluidics to realize controlled head immobilization in a semi-continuous fashion. To optimize the immobilization process, we characterize the hydrogel polymerization under different experimental conditions, including under the effect of fluid flow. We show that the Damköhler number specifically defined for our reactive transport phenomena can predict the success of such photopolymerized hydrogels used for sample immobilization. In addition to simultaneous recording of neural activity and behavior in C. elegans, we demonstrate our method's capability to temporarily reconfigure fluid flow and deliver chemical stimuli to the animal's nose to examine the animal's responses. We envision this approach to be useful for similar recordings for other small motile organisms, as well as scenarios where microfluidics and polymerization are used to control flow and rection.

The development of droplet-based microfluidic virus detection technology for human infectious diseases.

Virus-based human infectious diseases have a significant negative impact on people's health and social development. The need for quick, accurate, and early viral infection detection in preventive medicine is expanding. A microfluidic control is particularly suitable for point-of-care-testing virus diagnosis due to its advantages of low sample consumption, quick detection speed, simple operation, multi-functional integration, small size, and easy portability. It is also thought to have significant development potential and a wide range of application prospects in the research on virus detection technology. In an effort to aid researchers in creating novel microfluidic tools for virus detection, this review highlights recent developments of droplet-based microfluidics in virus detection research and also discusses the challenges and opportunities for rapid virus detection.

Alginate microgels encapsulation strategy of silver nanoparticles active against Candida albicans

Nanoparticles (NPs), offering high specific surface, are considered as the best potential anti-microbial agents for a wide range of medical nanotechnology. Among them, silver NPs (AgNPs) allow high biocidal activity. A design of ecological capsules is proposed where AgNPs of 22 nm in diameter are grafted on the surface of biobased nanocrystals obtained from cellulose (CNC) and chitin (ChiNC). These silver nanohybrids are dispersed in calcium-alginate microgels of 45–50 µm in diameter using microfluidic tools. Such double level of immobilization of AgNPs leads to highly stable carriers, prolongs shelf life and raises bioactivity, with a precise control of well dispersed AgNPs as determined by scanning transmission electron microscopy, UV-Vis spectroscopy and atomic absorption spectroscopy. Preliminary tests for antimicrobial activities of these new microgels have shown a significant inhibitory effect on Candida albicans, the most common fungal pathogen, responsible for thrush and vaginal yeast infections, for very low levels of silver.

Emerging Microfluidic Tools for Simultaneous Exosomes and Cargo Biosensing in Liquid Biopsy: New Integrated Miniaturized FFF-Assisted Approach for Colon Cancer Diagnosis

The early-stage diagnosis of cancer is a crucial clinical need. The inadequacies of surgery tissue biopsy have prompted a transition to a less invasive profiling of molecular biomarkers from biofluids, known as liquid biopsy. Exosomes are phospholipid bilayer vesicles present in many biofluids with a biologically active cargo, being responsible for cell-to-cell communication in biological systems. An increase in their excretion and changes in their cargo are potential diagnostic biomarkers for an array of diseases, including cancer, and they constitute a promising analyte for liquid biopsy. The number of exosomes released, the morphological properties, the membrane composition, and their content are highly related to the physiological and pathological states. The main analytical challenge to establishing liquid biopsy in clinical practice is the development of biosensors able to detect intact exosomes concentration and simultaneously analyze specific membrane biomarkers and those contained in their cargo. Before analysis, exosomes also need to be isolated from biological fluids. Microfluidic systems can address several issues present in conventional methods (i.e., ultracentrifugation, size-exclusion chromatography, ultrafiltration, and immunoaffinity capture), which are time-consuming and require a relatively high amount of sample; in addition, they can be easily integrated with biosensing systems. A critical review of emerging microfluidic-based devices for integrated biosensing approaches and following the major analytical need for accurate diagnostics is presented here. The design of a new miniaturized biosensing system is also reported. A device based on hollow-fiber flow field-flow fractionation followed by luminescence-based immunoassay is applied to isolate intact exosomes and characterize their cargo as a proof of concept for colon cancer diagnosis.

Brachytherapy-on-Chip: A Microfluidic Setup for In Vitro Interrogation of Hypoxic Spheroids

Despite evidence of its advantages in many cancers, brachytherapy (BT) remains clinically underused and understudied in the pre-clinical setting due to a lack of versatile RT-compatible in vitro tools that can emulate the tumor microenvironment and radiobiology of various cancers. Microfluidic devices use conventional cell culture methods in 3D-tumor models, are radiocompatible and can integrate radiobioassays. However, they are seldom used in pre-clinical BT research. This project engineered the first microfluidic tool for in vitro testing of BT, with applications in translational radiobiology. PDMS microfluidic chips were engineered to grow and culture concentric rows of hypoxic spheroids, and to allow the insertion of a clinical iodine-125 BT seed at the center of the device. FaDu (hypopharyngeal squamous cell carcinoma), SK-LMS-1 (leiomyosarcoma) and HCT116 (colorectal carcinoma) cell lines were selected for their clinical relevance and ability to form spheroids. Presence of hypoxia in spheroids was assessed by immunofluorescence (IF) staining for hypoxic protein CAIX. On-chip dose distribution was calculated using clinical TG-43 parameters and compared to EBT-XD radiographic film. Target dose criteria was fixed at 8 Gy in the center of the first row of spheroids. Treatment response was quantified by DNA damages (γH2AX IF, comet assay) and cell survival (clonogenic assay). Response of hypoxic and normoxic regions of spheroids will be compared in IF. A total of 15 spheroids that are 750µm or larger can be cultured in our device, arranged as 5 rows of 3 spread from 1.5mm to 7.5mm away from a central iodine-125 seed. 48h after cell seeding, hypoxic cores were observed in spheroids derived from FaDu (50 ± 4% of cross-section, N = 3) and SK-LMS-1 (46 ± 4% of cross-section, N = 3) cells, results are pending for HCT116. TG-43 formula predicts that the center of each row receives 8 Gy, 2.6 Gy, 1.2 Gy, 0.7 Gy and 0.4 Gy respectively. Radiographic film analysis confirms on-chip TG-43 calculated doses (N = 5, R2 = 0.999). Similarly, tail moment from comet assays follows predicted dose trends (n>60, N = 3). There was no statistical difference between 8 Gy BT and 8 Gy GammaCell (8 Gy BT vs 8 Gy GammaCell, p = 0.8758), with a statistical difference between 8 Gy BT (8 Gy BT vs 2.6 Gy BT,1.2 Gy BT,0.7 Gy BT,0.4 Gy BT,0 Gy, p < 0.0001), 2.6 Gy BT (2.6 Gy BT vs 1.2 Gy BT,0.7 Gy BT,0.4 Gy BT,0 Gy, p < 0.05) or 8 Gy GammaCell (8 Gy GammaCell vs 2.6 Gy BT,1.2 Gy BT,0.7 Gy BT,0.4 Gy BT,0 Gy, p < 0.0001) and other lower BT doses. For the first time, brachytherapy can be easily integrated on-chip and its effects evaluated on relevant 3D-tumor models. Our system allows simultaneous quantification of BT efficacy on normoxic and hypoxic cells treated at various doses. On-chip combination of BT with antitumor drug will be explored in future work. We hope this device will serve as further proof of the potential of BT/RT-on-chip systems for better drug development, treatment planification and theranostics.

Fast and reliable analysis of pH-responsive nanocarriers for drug delivery using microfluidic tools

During the last decades, there has been growing interest in the application of functionalized mesoporous nanomaterials as stimuli-responsive carriers for drug delivery. However, at present there is not a standardized methodology to evaluate their performance. The limitations of the different techniques reported in literature give rise to the necessity for new, simple, and cost-effective alternatives. This work constitutes a step forward in the development of advanced in vitro procedures for testing the behavior of nanocarriers, proposing a novel microfluidic platform. To test the capacity of the reported tool, the performance of amino-functionalized MCM-41 nanoparticles has been assessed. These materials show a pH-responsive mechanism, which prevents the drug release at acidic conditions, maximizing its distribution at neutral pH, thus, the selected release medium mimicked gastrointestinal conditions. As a first approximation, the delivery of Ru(bipy)32+ was evaluated, proving the advantages of the proposed microfluidic system: i) continuous flow of particles and media, ii) rigorous control of the residence time, temperature and pH, iii) enhanced mixing, iv) possibility to simulate different human body conditions and, v) possible integration with the continuous synthesis of nanocarriers. Finally, the microfluidic tool was used to analyze the delivery of the anti-inflammatory drug ibuprofen.

Microparticles with tunable, cell-like properties for quantitative acoustic mechanophenotyping

Mechanical properties of biological cells have been shown to correlate with their biomolecular state and function, and therefore methods to measure these properties at scale are of interest. Emerging microfluidic technologies can measure the mechanical properties of cells at rates over 20,000 cells/s, which is more than four orders of magnitude faster than conventional instrumentation. However, precise and repeatable means to calibrate and test these new tools remain lacking, since cells themselves are by nature variable. Commonly, microfluidic tools use rigid polymer microspheres for calibration because they are widely available in cell-similar sizes, but conventional microspheres do not fully capture the physiological range of other mechanical properties that are equally important to device function (e.g., elastic modulus and density). Here, we present for the first time development of monodisperse polyacrylamide microparticles with both tunable elasticity and tunable density. Using these size, elasticity, and density tunable particles, we characterized a custom acoustic microfluidic device that makes single-cell measurements of mechanical properties. We then applied the approach to measure the distribution of the acoustic properties within samples of human leukocytes and showed that the system successfully discriminates lymphocytes from other leukocytes. This initial demonstration shows how the tunable microparticles with properties within the physiologically relevant range can be used in conjunction with microfluidic devices for efficient high-throughput measurements of mechanical properties at single-cell resolution.