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Phenylketonuria (PKU) is characterized by an autosomal recessive mutation in the phenylalanine hydroxylase (PAH) gene. Impaired PAH enzyme activity leads to the accumulation of phenylalanine (Phe) and its metabolites in the bloodstream, which disrupts the central nervous system and causes psychomotor retardation. Early diagnosis of PKU is essential for timely intervention. Moreover, continuous monitoring of blood Phe levels is indispensable for prognosis, requiring a robust and reliable monitoring system. This study presents an automated lab-on-a-CD-based system for early diagnosis and monitoring of PKU treatment. This miniaturised system contains CD-shaped disposable cartridges, a mini centrifuge, and an electrochemical sensing unit. Modified screen-printed gold electrodes were used for the electrochemical measurements in cartridges. Electrode modification was conducted by electrochemical graphene oxide reduction and deposition on the electrode surface, which increased the sensitivity of the measurement 1.5 fold. The system used amperometric detection to measure Phe in the blood through oxidation of NAD + to NADH by the enzyme phenylalanine dehydrogenase. The limit of detection (LOD), limit of quantification (LOQ), and sensitivity of the system were 0.0524, 0.1587 mg dL -1 and 0.3338 μA mg -1 dL, respectively, within the 0-20 mg dL -1 measurement range ( R 2 = 0.9955). The performance of the lab-on-a-CD system was compared to the gold standard HPLC method. The accuracy was 83.1% for HPLC and 84.1% for the lab-on-a-CD system. In conclusion, this study successfully developed a portable diagnostic device for rapid (under 20 min), accurate and highly sensitive detection of Phe in whole blood.

The exceptional ability of liposomes to mimic a cellular lipid membrane makes them invaluable tools in biomembrane studies and bottom-up synthetic biology. Microfluidics provides a promising toolkit for creating giant liposomes in a controlled manner. Nevertheless, challenges associated with the microfluidic formation of double emulsions, as precursors to giant liposomes, limit the full exploration of this potential. In this study, we propose a PDMS-glass capillary hybrid device as a facile and versatile tool for the formation of double emulsions which not only eliminates the need for selective surface treatment, a well-known problem with PDMS formation chips, but also provides fabrication simplicity and reusability compared to the glass-capillary formation chips. These advantages make the presented device a versatile tool for forming double emulsions with varying sizes (spanning two orders of magnitude in diameter), shell thickness, number of compartments, and choice of solvents. We achieved robust thin shell double emulsion formation by operating the hybrid chip in double dripping mode without performing hydrophilic/phobic treatment a priori. In addition, as an alternative to the conventional, time-consuming density-based separation method, a tandem separation chip is developed to deliver double emulsions free of any oil droplet contamination in a continuous and rapid manner without any need for operator handling. The applicability of the device was demonstrated by forming giant liposomes using the solvent extraction method. This easy-to-replicate, flexible, and reliable microfluidic platform for the formation and separation of double emulsion templates paves the way for the high-throughput microfluidic generation of giant liposomes and synthetic cells, opening exciting avenues for biomimetic research. The presented giant liposome assembly line features a novel treatment-free hybrid chip for double emulsion formation coupled with a high throughput separation chip for sample purification., Competing Interests: Conflict of interest: The authors declare no competing interests., (© 2024. The Author(s).)

Organoids are emerging as a powerful tool to investigate complex biological structures in vitro . Vascularization of organoids is crucial to recapitulate the morphology and function of the represented human organ, especially in the case of the kidney, whose primary function of blood filtration is closely associated with blood circulation. Current in vitro microfluidic approaches have only provided initial vascularization of kidney organoids, whereas in vivo transplantation to animal models is problematic due to ethical problems, with the exception of xenotransplantation onto a chicken chorioallantoic membrane (CAM). Although CAM can serve as a good environment for vascularization, it can only be used for a fixed length of time, limited by development of the embryo. Here, we propose a novel lab on a chip design that allows organoids of different origin to be cultured and vascularized on a CAM, as well as to be transferred to in vitro conditions when required. Mouse embryonic kidneys cultured on the CAM showed enhanced vascularization by intrinsic endothelial cells, and made connections with the chicken vasculature, as evidenced by blood flowing through them. After the chips were transferred to in vitro conditions, the vasculature inside the organoids was successfully maintained. To our knowledge, this is the first demonstration of the combination of in vivo and in vitro approaches applied to microfluidic chip design.

Plant-derived nanovesicles such as bilberries nanovesicles (BNVs) show immense promise as next-generation biotherapeutics and functional food ingredients; however, their isolation, purification, and storage on a large scale remain a challenge. In this study, biocompatible and nanostructured composite all-cellulose membranes are introduced as a scalable and straightforward approach to the isolation of BNV. The membranes consisting of a cellulose acetate matrix infused with anionic or cationic nanocelluloses promoted selective capture of BNVs through electrostatic and size-exclusion-mediated depth filtration. Furthermore, the surface of the composite membrane acted as a storage matrix for BNVs, ensuring their prolonged stability at 4 °C. The BNVs stored in the membrane could be promptly released through elution assisted by low-pressure vacuum filtration or diffusion in liquid media. The morphology, bioactivity, and stability of the extracted BNVs were preserved, and the release rate of BNVs in different cell cultures could be regulated, facilitating their use for local therapy. Consequently, this approach paves the way for the scalable production, purification, and storage of nanovesicles and advances their use in biotherapeutics and functional foods.

Competing Interests: Competing interests statement:U.A.G. and Case Western Reserve University have financial interests in Hemex Health Inc. E.K., U.A.G., and Case Western Reserve University have financial interests in BioChip Labs Inc. U.A.G. and Case Western Reserve University have financial interests in Xatek Inc. U.A.G. has financial interests in DxNow Inc. Financial interests include licensed intellectual property, stock ownership, research funding, employment, and consulting. Hemex Health Inc. offers point-of-care diagnostics for hemoglobin disorders, anemia, and malaria. BioChip Labs Inc. offers commercial clinical microfluidic biomarker assays for inherited or acquired blood disorders. Xatek Inc. offers point-of-care global assays to evaluate the hemostatic process. DxNow Inc. offers microfluidic and bio-imaging technologies for in vitro fertilization, forensics, and diagnostics. Competing interests of Case Western Reserve University employees are overseen and managed by the Conflict of Interests Committee according to a Conflict-of-Interest Management Plan. The study was conducted prior to E.K. employment at IDEXX Laboratories, and the ideas expressed are not in his capacity as an IDEXX employee. A.D. serves on the Scientific Advisory Board of Hemanext Inc. and Macopharma Inc., and his competing interests are managed by the University of Colorado in accordance with their conflict-of-interest policies. S.N.T. and M.T. serve on the Scientific Advisory Board of Sylvatica Biotech Inc., a company focused on developing high subzero organ preservation technology. Competing interests for all authors affiliated with the Massachusetts General Hospital are managed by the MGH and Mass General Brigham in accordance with their conflict-of-interest policies. The remaining authors declare no competing interests.

Transfusion of red blood cells (RBCs) is one of the most valuable and widespread treatments in modern medicine. Lifesaving RBC transfusions are facilitated by the cold storage of RBC units in blood banks worldwide. Currently, RBC storage and subsequent transfusion practices are performed using simplistic workflows. More specifically, most blood banks follow the "first-in-first-out" principle to avoid wastage, whereas most healthcare providers prefer the "last-in-first-out" approach simply favoring chronologically younger RBCs. Neither approach addresses recent advances through -omics showing that stored RBC quality is highly variable depending on donor-, time-, and processing-specific factors. Thus, it is time to rethink our workflows in transfusion medicine taking advantage of novel technologies to perform RBC quality assessment. We imagine a future where lab-on-a-chip technologies utilize novel predictive markers of RBC quality identified by -omics and machine learning to usher in a new era of safer and precise transfusion medicine.

The isolation of extracellular vesicles (EVs) from milk, a complex mixture of colloidal structures having a comparable size to EVs, is challenging. Although ultracentrifugation (UC) has been widely used for EV isolation, this has significant limitations, including a long processing time at high g-force conditions and large sample volume requirements. We introduced a new approach based on nature nanoentities cellulose nanofibers (CNFs) and short time and low g-force centrifugation to isolate EVs from various milk fractions. The flexible and entangled network of CNFs forms nanoporous, which entraps the EVs. Further, positively charged CNFs interact with anionic EVs through an electrostatic attraction, promoting their isolation with efficiency comparable with UC. The functionality and toxicity of isolated milk EVs were tested in Caco2 cells. Overall, the newly developed approach provides straightforward isolation and biocompatibility and preserves the natural properties of the isolated EVs, enabling further applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Extracellular nanovesicles, particularly exosomes, can deliver their diverse bioactive biomolecular content, including miRNAs, proteins, and lipids, thus providing a context for investigating the capability of exosomes to induce stem cells toward lineage-specific cells and tissue regeneration. In this study, it is demonstrated that rat subventricular zone neural stem cell-derived exosomes (rSVZ-NSCExo) can control neural-lineage specification of human mesenchymal stem cells (hMSCs). Microarray analysis shows that the miRNA content of rSVZ-NSCExo is a faithful representation of rSVZ tissue. Through immunocytochemistry, gene expression, and multi-omics analyses, the capability to use rSVZ-NSCExo to induce hMSCs into a neuroglial or neural stem cell phenotype and genotype in a temporal and dose-dependent manner via multiple signaling pathways is demonstrated. The current study presents a new and innovative strategy to modulate hMSCs fate by harnessing the molecular content of exosomes, thus suggesting future opportunities for rSVZ-NSCExo in nerve tissue regeneration., (© 2022 Wiley-VCH GmbH.)

Low proliferation capacity of corneal endothelial cells (CECs) and worldwide limitations in transplantable donor tissues reveal the critical need of a robust approach for in vitro CEC growth. However, preservation of CEC-specific phenotype with increased proliferation has been a great challenge. Here we offer a biomimetic cell substrate design, by optimizing mechanical, topographical and biochemical characteristics of materials with CEC microenvironment. We showed the surprising similarity between topographical features of white rose petals and corneal endothelium due to hexagonal cell shapes and physiologically relevant cell density (≈ 2000 cells/mm 2 ). Polydimethylsiloxane (PDMS) substrates with replica of white rose petal topography and cornea-friendly Young's modulus (211.85 ± 74.9 kPa) were functionalized with two of the important corneal extracellular matrix (ECM) components, collagen IV (COL 4) and hyaluronic acid (HA). White rose petal patterned and COL 4 modified PDMS with optimized stiffness provided enhanced bovine CEC response with higher density monolayers and increased phenotypic marker expression. This biomimetic approach demonstrates a successful platform to improve in vitro cell substrate properties of PDMS for corneal applications, suggesting an alternative environment for CEC-based therapies, drug toxicity investigations, microfluidics and organ-on-chip applications., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Corneal endothelial cells (CECs) have limited proliferation ability leading to corneal endothelium (CE) dysfunction and eventually vision loss when cell number decreases below a critical level. Although transplantation is the main treatment method, donor shortage problem is a major bottleneck. The transplantation of in vitro developed endothelial cells with desirable density is a promising idea. Designing cell substrates that mimic the native CE microenvironment is a substantial step to achieve this goal. In the presented study, we prepared polyacrylamide (PA) cell substrates that have a microfabricated topography inspired by the dimensions of CECs. Hydrogel surfaces were prepared via two different designs with small and large patterns. Small patterned hydrogels have physiologically relevant hexagon densities (∼2000 hexagons/mm 2 ), whereas large patterned hydrogels have sparsely populated hexagons (∼400 hexagons/mm 2 ). These substrates have similar elastic modulus of native Descemet's membrane (DM; ∼50 kPa) and were modified with Collagen IV (Col IV) to have biochemical content similar to native DM. The behavior of bovine corneal endothelial cells on these substrates was investigated and results show that cell proliferation on small patterned substrates was significantly (p = 0.0004) higher than the large patterned substrates. Small patterned substrates enabled a more densely populated cell monolayer compared to other groups (p = 0.001 vs. flat and p < 0.0001 vs. large patterned substrates). These results suggest that generating bioinspired surface topographies augments the formation of CE monolayers with the desired cell density, addressing the in vitro development of CE layers., (© 2021 John Wiley & Sons Ltd.)

Simple, low-cost, robust, and scalable fabrication of microscopic linear barcodes with high levels of complexity and multiple authentication layers is critical for emerging applications in information security and anti-counterfeiting. This manuscript presents a novel approach for fabrication of microscopic linear barcodes that can be visualized under Raman microscopy. Microfluidic channels are used as molds to generate linear patterns of end-grafted polymers on a substrate. These patterns serve as templates for area-selective binding of colloidal gold nanoparticles resulting in plasmonic arrays. The deposition of multiple taggant molecules on the plasmonic arrays via a second microfluidic mold results in a linear barcode with unique Raman fingerprints that are enhanced by the underlying plasmonic nanoparticles. The width of the bars is as small as 10 μm, with a total barcode length on the order of 100 μm. The simultaneous use of geometric and chemical security layers provides a high level of complexity challenging the counterfeiting of the barcodes. The additive, scalable, and inexpensive nature of the presented approach can be easily adapted to different colloidal nanomaterials and applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The flow behavior of blood is determined mainly by red blood cell (RBC) deformation and aggregation as well as blood viscoelasticity. These intricately interdependent parameters should be monitored by healthcare providers to understand all aspects of circulatory flow dynamics under numerous cases including cardiovascular and infectious diseases. Current medical instruments and microfluidic systems lack the ability to quantify these parameters all at once and in physiologically relevant flow conditions. This work presents a handheld platform and a measurement method for quantitative analysis of multiple of these parameters from 50 μl undiluted blood inside a miniaturized channel. The assay is based on an optical transmission analysis of collective RBC deformation and aggregation under near-infrared illumination during a 1 s damped oscillatory flow and at stasis, respectively. Measurements with blood of different hemo-rheological properties demonstrate that the presented approach holds a potential for initiating simultaneous and routine on-chip blood flow analysis even in resource-poor settings.

Under the simultaneous use of pressure-driven flow and DC electric field, migration of particles inside microfluidic channels exhibits intricate focusing dynamics. Available experimental and analytical studies fall short in giving a thorough explanation to particle equilibrium states. Also, the understanding is so far limited to the results based on Newtonian and neutral viscoelastic carrier fluids. Hence, a holistic approach is taken in this study to elaborate the interplay of governing electrophoretic and slip-induced/elastic/shear gradient lift forces. First, we carried out experimental studies on particle migration in Newtonian, neutral viscoelastic, and polyelectrolyte viscoelastic media to provide a comprehensive understanding of particle migration. The experiments with the viscoelastic media led to contradictory results with the existing explanations. Then, we introduced the Electro-Viscoelastic Migration (EVM) theory to give a unifying explanation to particle migration in Newtonian and viscoelastic solutions. Confocal imaging with fluorescent-labeled polymer solutions was used to explore the underlying migration behavior. A surprising outcome of our results is the formation of cross-sectionally nonuniform viscoelasticity that may have unique applications in microfluidic particle focusing.

Droplet-based microfluidic systems require a precise control of droplet physical properties; hence, measuring the morphological properties of droplets is critical to obtain high sensitivity analysis. The ability to perform such measurements in real-time is another demand which has not been addressed yet. In this study, we used coplanar electrodes configured in the differential measurement mode for impedimetric measurement of size and velocity. To obtain the size of the droplets, detailed 3D finite element simulations of the system were performed. The interaction of the non-uniform electric field and the droplet was investigated. Electrode geometry optimization steps were described and design guideline rules were laid out. User-friendly software was developed for real-time observation of droplet length and velocity together with in situ statistical analysis results. A comparison between impedimetric and optical measurement tools is given. Finally, to illustrate the benefit of having real-time analysis, iDM was used to synthesize particles with a predefined monodispersity limit and to study the response times of syringe pump and pressure pump driven droplet generation devices. This analysis allows one to evaluate the 'warm-up' time for a droplet generator system, after which droplets reach the desired steady-state size required by the application of interest.

Continuous development of security features is mandatory for the fight against forgery of valuable documents and products, the most notable example being banknotes. Such features demonstrate specific properties under certain stimuli such as fluorescent patterns glowing under ultraviolet light. These security features should also be hard to copy by unlicensed people and be interrogated by anyone using easily accessible tools. To this end, this Letter demonstrates the development of an ideal security feature enabled by the realization of modular metamaterials based on metal-dielectric-metal cavities that consist of two separate parts: metal nanoparticles on an elastomeric substrate and a bottom mirror coated with a thin dielectric. Patterns generated by creating nanometer-thick changes in the dielectric layer are invisible (encrypted) and can only be detected (decrypted) by sticking the elastomeric patch on. The observed optical effects such as visibility and colors can only be produced with the correct combination of materials and film thicknesses, making the proposed structures a strong alternative to compromised security features.

Elastic nature of the viscoelastic fluids induces lateral migration of particles into a single streamline and can be used by microfluidic based flow cytometry devices. In this study, we investigated focusing efficiency of polyethylene oxide based viscoelastic solutions at varying ionic concentration to demonstrate their use in impedimetric particle characterization systems. Rheological properties of the viscoelastic fluid and particle focusing performance are not affected by ionic concentration. We investigated the viscoelastic focusing dynamics using polystyrene (PS) beads and human red blood cells (RBCs) suspended in the viscoelastic fluid. Elasto-inertial focusing of PS beads was achieved with the combination of inertial and viscoelastic effects. RBCs were aligned along the channel centerline in parachute shape which yielded consistent impedimetric signals. We compared our impedance-based microfluidic flow cytometry results for RBCs and PS beads by analyzing particle transit time and peak amplitude at varying viscoelastic focusing conditions obtained at different flow rates. We showed that single orientation, single train focusing of nonspherical RBCs can be achieved with polyethylene oxide based viscoelastic solution that has been shown to be a good candidate as a carrier fluid for impedance cytometry., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

We introduce a droplet-based biomolecular detection platform using robust, versatile, and low-cost superhydrophilic patterned superhydrophobic surfaces. Benefitting from confinement and evaporation-induced shrinkage of droplets on wetted patterns, we show enrichment-based biomolecular detection using very low sample volumes. First, we developed a glucose assay using fluorescent polydopamine (PDA) based on enhancement of PDA emission by hydrogen peroxide (H 2 O 2 ) produced in enzyme-mediated glucose oxidation reaction. Incubation in evaporating droplets resulted in brighter fluorescence compared to that in bulk solutions. Droplet assay was highly sensitive toward increasing glucose concentration while that in milliliter-volume solutions resulted in no fluorescence enhancement at similar time scales. This is due to droplet evaporation that increased the reaction rate by causing enrichment of PDA and glucose/glucose oxidase as well as increased concentration of H 2 O 2 generated in shrinking droplet. Second, we chemically functionalized wetted patterns with single-stranded DNA and developed fluorescence-based DNA detection to demonstrate the adaptability of the patterned surfaces for a different class of assay. We achieved detection of glucose and DNA with concentration down to 130 μM and 200 fM, respectively. Patterned superhydrophobic surfaces with their simple production, sensitive response, and versatility present potential for bioanalysis from low sample volumes., Competing Interests: The authors declare no competing financial interest.

The process of detection and separation of yeast cells based on their morphological characteristics is critical to the understanding of cell division cycles, which is of vital importance to the understanding of some diseases such as cancer. The traditional process of manual detection is usually tedious and inconsistent. This paper presents a microfluidic device integrated with microvalves for fluid control for the sorting of yeast cells using image processing algorithms and confirmation based on their fluorescent tag. The proposed device is completely automated, low cost and easy to implement in an academic research setting. Design details of the integrated microfluidic system are highlighted in this paper, along with experimental validation. Real time cell sorting was demonstrated with a cell detection rate of 12 cells per minute.

Microflow cytometry is a powerful technique for characterization of particles suspended in a solution. In this work, we present a microflow cytometer based on viscoelastic focusing. 3D single-line focusing of microparticles was achieved in a straight capillary using viscoelastic focusing which alleviated the need for sheath flow or any other actuation mechanism. Optical detection was performed by fiber coupled light source and photodetectors. Using this system, we present the detection of microparticles suspended in three different viscoelastic solutions. The rheological properties of the solutions were measured and used to assess the focusing performance both analytically and numerically. The results were verified experimentally, and it has been shown that polyethlyene oxide (PEO) and hyaluronic acid (HA) based sheathless microflow cytometer demonstrates similar performance to state-of-the art flow cytometers. The sheathless microflow cytometer was shown to present 780 particles/s throughput and 5.8% CV for the forward scatter signal for HA-based focusing. The presented system is composed of a single capillary to accommodate the fluid and optical fibers to couple the light to the fluid of interest. Thanks to its simplicity, the system has the potential to widen the applicability of microflow cytometers.

The erythrocyte sedimentation rate (ESR) is a frequently used 30 min or 60 min clinical test for screening of several inflammatory conditions, infections, trauma, and malignant diseases, as well as non-inflammatory conditions including prostate cancer and stroke. Erythrocyte aggregation (EA) is a physiological process where erythrocytes form face-to-face linear structures, called rouleaux, at stasis or low shear rates. In this work, we proposed a method for ESR measurement from EA. We developed a microfluidic opto-electro-mechanical system, using which we experimentally showed a significant correlation (R 2 = 0.86) between ESR and EA. The microfluidic system was shown to measure ESR from EA using fingerprick blood in 2 min. 40 μl of whole blood is filled in a disposable polycarbonate cartridge which is illuminated with a near infrared emitting diode. Erythrocytes were disaggregated under the effect of a mechanical shear force using a solenoid pinch valve. Following complete disaggregation, transmitted light through the cartridge was measured using a photodetector for 1.5 min. The intensity level is at its lowest at complete disaggregation and highest at complete aggregation. We calculated ESR from the transmitted signal profile. We also developed another microfluidic cartridge specifically for monitoring the EA process in real-time during ESR measurement. The presented system is suitable for ultrafast, low-cost, and low-sample volume measurement of ESR at the point-of-care.

Droplet-based microfluidic systems offer tremendous benefits for high throughput biochemical assays. Despite the wide use of electrical detection for microfluidic systems, application of impedimetric sensing for droplet systems is very limited. This is mainly due to the insulating oil-based continuous phase used for most aqueous samples of interest. We present modelling and experimental verification of impedimetric detection of hemagglutination in microdroplets. We have detected agglutinated red blood cells in microdroplets and screened whole blood samples for multiple antibody sera using conventional microelectrodes. We were able to form antibody and whole blood microdroplets in PDMS microchannels without any tedious chemical surface treatment. Following the injection of a blood sample into antibody droplets, we have detected the agglutination-positive and negative droplets in an automated manner. In order to understand the characteristics of impedimetric detection inside microdroplets, we have developed the lumped electrical circuit equivalent of an impedimetric droplet content detection system. The empirical lumped element values are in accordance with similar models developed for single phase electrical impedance spectroscopy systems. The presented approach is of interest for label-free, quantitative analysis of droplets. In addition, the standard electronic equipment used for detection allows miniaturized detection circuitries that can be integrated with a fluidic system for a quantitative microdroplet-based hemagglutination assay that is conventionally performed in well plates.

This paper proposes a novel method for measuring blood plasma and serum viscosity with a microcantilever-based MEMS sensor. MEMS cantilevers are made of electroplated nickel and actuated remotely with magnetic field using an electro-coil. Real-time monitoring of cantilever resonant frequency is performed remotely using diffraction gratings fabricated at the tip of the dynamic cantilevers. Only few nanometer cantilever deflection is sufficient due to interferometric sensitivity of the readout. The resonant frequency of the cantilever is tracked with a phase lock loop (PLL) control circuit. The viscosities of liquid samples are obtained through the measurement of the cantilever's frequency change with respect to a reference measurement taken within a liquid of known viscosity. We performed measurements with glycerol solutions at different temperatures and validated the repeatability of the system by comparing with a reference commercial viscometer. Experimental results are compared with the theoretical predictions based on Sader's theory and agreed reasonably well. Afterwards viscosities of different Fetal Bovine Serum and Bovine Serum Albumin mixtures are measured both at 23°C and 37°C, body temperature. Finally the viscosities of human blood plasma samples taken from healthy donors are measured. The proposed method is capable of measuring viscosities from 0.86 cP to 3.02 cP, which covers human blood plasma viscosity range, with a resolution better than 0.04 cP. The sample volume requirement is less than 150 μl and can be reduced significantly with optimized cartridge design. Both the actuation and sensing are carried out remotely, which allows for disposable sensor cartridges., (Copyright © 2013. Published by Elsevier Inc.)

This is the second part of a two-part study on the generation of droplets at a microfluidic T-junction operating in the transition regime. In the preceding paper [Phys. Rev. E 85, 016322 (2012)], we presented our experimental observations of droplet formation and decomposed the process into three sequential stages defined as the lag, filling, and necking stages. Here we develop a model that describes the performance of microfluidic T-junction generators working in the squeezing to transition regimes where confinement of the droplet dominates the formation process. The model incorporates a detailed geometric description of the drop shape during the formation process combined with a force balance and necking criteria to define the droplet size, production rate, and spacing. The model inherently captures the influence of the intersection geometry, including the channel width ratio and height-to-width ratio, capillary number, and flow ratio, on the performance of the generator. The model is validated by comparing it to speed videos of the formation process for several T-junction geometries across a range of capillary numbers and viscosity ratios.

This is the first part of a two-part study on the generation of droplets at a microfluidic T-junction operating in the transition regime where confinement of the droplet creates a large squeezing pressure that influences droplet formation. In this regime, the operation of the T-junction depends on the geometry of the intersection (height-to-width ratio, inlet width ratio), capillary number, flow ratio, and viscosity ratio of the two phases. Here in paper I we presented our experimental observations through the analysis of high-speed videos of the droplet formation process. Various parameters are tracked during the formation cycle such as the shape of the droplet (penetration depth and neck), interdroplet spacing, production rate, and flow of both phases across several T-junction designs and flow conditions. Generally, the formation process is defined by a two-stage model consisting of an initial filling stage followed by a necking stage. However, video evidence suggests the inclusion of a third stage, which we term the lag stage, at the beginning of the formation process that accounts for the retraction of the interface back into the injection channel after detachment. Based on the observations made in this paper, a model is developed to describe the formation process in paper II, which can be used to understand the design and operation of T-junction generators in the transition regime.

When droplets enter a junction they sort to the channel with the highest flow rate at that instant. Transport is regulated by a discrete time-delayed feedback that results in a highly periodic behavior where specific patterns can continue to cycle indefinitely. Between these highly ordered regimes are chaotic structures where no pattern is evident. Here we develop a model that describes droplet sorting under various asymmetries: branch geometry (length, cross-section), droplet resistance and pressures. First, a model is developed based on the continuum assumption and then, with the assistance of numerical simulations, a discrete model is derived to predict the length and composition of the sorting pattern. Furthermore we derive all unique sequences that are possible for a given distribution and develop a preliminary estimation of why chaotic regimes form. The model is validated by comparing it to numerical simulations and results from microfluidic experiments in PDMS chips with good agreement.

The study of yeast cell morphology requires consistent identification of cell cycle phases based on cell bud size. A computer-based image processing algorithm is designed to automatically classify microscopic images of yeast cells in a microfluidic channel environment. The images were enhanced to reduce background noise, and a robust segmentation algorithm is developed to extract geometrical features including compactness, axis ratio, and bud size. The features are then used for classification, and the accuracy of various machine-learning classifiers is compared. The linear support vector machine, distance-based classification, and k-nearest-neighbor algorithm were the classifiers used in this experiment. The performance of the system under various illumination and focusing conditions were also tested. The results suggest it is possible to automatically classify yeast cells based on their morphological characteristics with noisy and low-contrast images.

A fully integrated polydimethylsiloxane (PDMS)/modified PDMS membrane/SU-8/quartz hybrid chip was developed for protein separation using isoelectric focusing (IEF) mechanism coupled with whole-channel imaging detection (WCID) method. This microfluidic chip integrates three components into one single chip: (i) modified PDMS membranes for separating electrolytes in the reservoirs from the sample in the microchannel and thus reducing pressure disturbance, (ii) SU-8 optical slit to block UV light (below 300 nm) outside the channel aiming to increase detection sensitivity, and (iii) injection and discharge capillaries for continuous operation. Integration of all these components on a single chip is challenging because it requires fabrication techniques for perfect bonding between different materials and is prone to leakage and blockage. This study has addressed all the challenges and presented a fully integrated chip, which is more robust with higher sensitivity than the previously developed IEF chips. This chip was tested by performing protein and pI marker separation. The separation results obtained in this chip were compared with that obtained in commercial cartridges. Side-by-side comparison validated the developed chip and fabrication techniques., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This study presents a microfluidic system that incorporates electroosmotic pumps, a concentration gradient generator and a fish cell line (rainbow trout gill) to perform toxicity testing on fish cells seeded in the system. The system consists of three mechanical components: (1) a toxicity testing chip containing a microfluidic gradient generator which creates a linear concentration distribution of toxicant in a cell test chamber, (2) an electroosmotic (EO) pump chip that controls the flow rate and operation of the toxicity chip, and (3) indirect reservoirs that connect the two chips allowing for the toxicant solution to be pumped separately from the electroosmotic pump solution. The flow rate and stability of the EO pumps was measured and tested by monitoring the gradient generator using fluorescence microscopy. Furthermore, a lethality test was performed with this system setup using a rainbow trout gill cell line (RTgill-W1) as the test cells and sodium dodecyl sulfate as a model toxicant. A gradient of sodium dodecyl sulfate, from 0 to 50 microg mL(-1), was applied for 1 hr to the attached cells, and the results were quantified using a Live/Dead cell assay. This work is a preliminary study on the application of EO pumps in a living cell assay, with the potential to use the pumps in portable water quality testing devices with RTgill-W1 cells as the biosensors.