Algae Adhesion onto Silicone is Sensitive to Environment-Induced Surface Restructuring.

Resistance to algae contamination is an important characteristic of insulators used in overhead power distribution in coastal environments. It is therefore important to understand the parameters governing algae adhesion onto polymer insulator materials such as silicone. Flow cell-based shear experiments were conducted in order to characterize the adhesion strength of algae onto polydimethylsiloxane surfaces, comparing fresh polymer substrates with those that have been soaked in water and saline solutions for 1 month. Both freshwater algae and seawater species could withstand considerably less drag force and were therefore more easily removed when the polymer was soaked in salt water. The polymer surface was found to be unaltered in terms of its roughness, contact angle, and lack of water uptake; no macroscopic surface characterization was therefore able to account for the differences in cell adhesion strength resulting from the soaking treatment. Surface-specific nonlinear vibrational spectroscopy, however, revealed subtle differences in the orientation of surface methyl groups that resulted from the water and saline exposure.

The role of temperature in the formation of human-mimetic artificial cell membranes using droplet interface bilayers (DIBs).

Droplet interface bilayers (DIBs) have recently started to be used as human-mimetic artificial cell membranes. DIBs are bilayer sections created at the interface of two aqueous droplets, such that one droplet can be used as a donor compartment and the other as an acceptor compartment for the quantification of molecular transport across the artificial cell membrane. However, synthetic phospholipids are overwhelmingly used to create DIBs instead of naturally derived phospholipids, even though the diverse distribution of phospholipids in the latter is more biomimetic. We present the first systematic study of the role of temperature in DIB formation, which shows that the temperature at which DIBs are formed is a key parameter for the formation of DIBs using naturally derived phospholipids in a microfluidic platform. The phospholipids that are most abundant in mammalian cell membranes (phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositol (PI)) only form DIBs when the temperature is above the phase transition temperature (Tm). Similarly, DIB formation usually only occurs above the highest Tm of a single phospholipid in a bespoke formulation. In addition, we show a new phenomenon wherein the DIB "melts" without disintegrating for bilayers formed predominantly of phospholipids that occupy cylindrical spaces. We also demonstrate differences in DIB formation rates as well as permeability of these biomimetic membranes. Given the difficulties associated with making DIBs using naturally derived phospholipids, we anticipate this work will illuminate the role of phospholipid phase transition in mono- and bilayer formation and lay the foundation for DIBs to be used as human-mimetic artificial cell membranes.

Microfluidic Technique for the Simultaneous Quantification of Emulsion Instabilities and Lipid Digestion Kinetics.

Quantifying the impact of environmental physicochemical changes on the microstructure of lipid delivery systems is challenging. Therefore, we have developed a methodology to quantify the coalescence of oil-in-water emulsion droplets during lipid digestion in situ on a single droplet level. This technique involves a custom-made glass microfluidic platform, in which oil droplets can be trapped as single droplets, or several droplets per trap. The physicochemical environment can be controlled, and droplet digestion, as well as coalescence, can be visualized. We show that the exchange of the physicochemical conditions in the entire reaction chamber can be reached in under 30 s. Microparticle image velocimetry allowed mapping of the flow profile and demonstrated the tuneability of the shear profile in the device. The extraction of quantitative information regarding the physical characteristics of the droplets during digestion was performed using an automated image analysis throughout the digestion process. Therefore, we were able to show that oil-in-water emulsions stabilized by proteins coalesced under human gastric conditions. This coalescence delayed the overall lipid digestion kinetics. The droplets that coalesced during digestion were hydrolyzed 1.4 times slower than individually trapped droplets. Thus, the microstructural evolution of lipid delivery systems is a crucial factor in lipid digestion kinetics. This novel technique allows the simultaneous quantification of the impact that the physicochemical environment has on both the lipid droplet microstructure and the lipid release patterns.

A Microfluidic Platform for the Rapid Determination of Distribution Coefficients by Gravity-Assisted Droplet-Based Liquid-Liquid Extraction.

The determination of pharmacokinetic properties of drugs, such as the distribution coefficient (D) is a crucial measurement in pharmaceutical research. Surprisingly, the conventional (gold standard) technique used for D measurements, the shake-flask method, is antiquated and unsuitable for the testing of valuable and scarce drug candidates. Herein, we present a simple microfluidic platform for the determination of distribution coefficients using droplet-based liquid-liquid extraction. For simplicity, this platform makes use of gravity to enable phase separation for analysis and is 48 times faster and uses 99% less reagents than performing an equivalent measurement using the shake-flask method. Furthermore, the D measurements achieved in our platform are in good agreement with literature values measured using traditional shake-flask techniques. Since D is affected by volume ratios, we use the apparent acid dissociation constant, pK', as a proxy for intersystem comparison. Our platform determines a pK' value of 7.24 ± 0.15, compared to 7.25 ± 0.58 for the shake-flask method in our hands and 7.21 for the shake-flask method in the literature. Devices are fabricated using injection molding, the batchwise fabrication time is <2 min per device (at a cost of $1 U.S. per device), and the interdevice reproducibility is high.

Droplet-based microfluidic platform for high-throughput, multi-parameter screening of photosensitizer activity.

We present a fully integrated droplet-based microfluidic platform for the high-throughput assessment of photodynamic therapy photosensitizer (PDT) efficacy on Escherichia coli. The described platform is able to controllably encapsulate cells and photosensitizer within pL-volume droplets, incubate the droplets over the course of several days, add predetermined concentrations of viability assay agents, expose droplets to varying doses of electromagnetic radiation, and detect both live and dead cells online to score cell viability. The viability of cells after encapsulation and incubation is assessed in a direct fashion, and the viability scoring method is compared to model live/dead systems for calibration. Final results are validated against conventional colony forming unit assays. In addition, we show that the platform can be used to perform concurrent measurements of light and dark toxicity of the PDT agents and that the platform allows simultaneous measurement of experimental parameters that include dark toxicity, photosensitizer concentration, light dose, and oxygenation levels for the development and testing of PDT agents.

Challenges and opportunities in achieving the full potential of droplet interface bilayers.

Model membranes can be used to elucidate the intricacies of the chemical processes that occur in cell membranes, but the perfectly biomimetic, yet bespoke, model membrane has yet to be built. Droplet interface bilayers are a new type of model membrane able to mimic some features of real cell membranes better than traditional models, such as liposomes and black lipid membranes. In this Perspective, we discuss recent work in the field that is starting to showcase the potential of these model membranes to enable the quantification of membrane processes, such as the behaviour of protein transporters and the prediction of in vivo drug movement, and their use as scaffolds for electrophysiological measurements. We also highlight the challenges that remain to enable droplet interface bilayers to achieve their full potential as artificial cells, and as biological analytical platforms to quantify molecular transport.

Materials and methods for droplet microfluidic device fabrication.

Since the first reports two decades ago, droplet-based systems have emerged as a compelling tool for microbiological and (bio)chemical science, with droplet flow providing multiple advantages over standard single-phase microfluidics such as removal of Taylor dispersion, enhanced mixing, isolation of droplet contents from surfaces, and the ability to contain and address individual cells or biomolecules. Typically, a droplet microfluidic device is designed to produce droplets with well-defined sizes and compositions that flow through the device without interacting with channel walls. Successful droplet flow is fundamentally dependent on the microfluidic device - not only its geometry but moreover how the channel surfaces interact with the fluids. Here we summarise the materials and fabrication techniques required to make microfluidic devices that deliver controlled uniform droplet flow, looking not just at physical fabrication methods, but moreover how to select and modify surfaces to yield the required surface/fluid interactions. We describe the various materials, surface modification techniques, and channel geometry approaches that can be used, and give examples of the decision process when determining which material or method to use by describing the design process for five different devices with applications ranging from field-deployable chemical analysers to water-in-water droplet creation. Finally we consider how droplet microfluidic device fabrication is changing and will change in the future, and what challenges remain to be addressed in the field.

Investigating the effect of phospholipids on droplet formation and surface property evolution in microfluidic devices for droplet interface bilayer (DIB) formation.

Despite growing interest in droplet microfluidic methods for droplet interface bilayer (DIB) formation, there is a dearth of information regarding how phospholipids impact device function. Limited characterization has been carried out for phospholipids, either computationally (in silico) or experimentally (in situ) in polydimethylsiloxane (PDMS) microfluidic devices, despite recent work providing a better understanding of how other surfactants behave in microfluidic systems. Hence, microfluidic device design for DIB applications relies heavily on trial and error, with many assumptions made about the impact of phospholipids on droplet formation and surface properties. Here, we examine the effects of phospholipids on interfacial tension, droplet formation, wetting, and hence device longevity, using DPhPC as the most widely used lipid for DIB formation. We use a customized COMSOL in silico model in comparison with in situ experimental data to establish that the stabilization of droplet formation seen when the lipid is dosed in the aqueous phase (lipid-in) or in the oil phase (lipid-out) is directly dependent on the effects of lipids on the device surface properties, rather than on how fast they coat the droplet. Furthermore, we establish a means to visually characterize surface property evolution in the presence of lipids and explore rates of device failure in the absence of lipid, lipid-out, and lipid-in. This first exploration of the effects of lipids on device function may serve to inform the design of microfluidic devices for DIB formation as well as to troubleshoot causes of device failure during microfluidic DIB experiments.

Microfluidic technologies for drug discovery and development: friend or foe?

There is a point in the evolution of every new technology when questions need to be asked regarding its usefulness and impact. Although microfluidic technologies have drastically decreased the scales at which laboratory processes can be performed and have enabled scientific advances that would have otherwise not been possible, it is time to consider whether these technologies are more disruptive than enabling. Here, my aims are to introduce researchers in the broad fields of drug discovery and development to the advantages and disadvantages of microfluidic technologies, to highlight current work showing how microfluidic technologies can be used at different stages in the drug discovery and development process, to discuss how we can transfer academic breakthroughs in the field of microfluidic technologies to industrial environments, and to examine whether microfluidic technologies have the potential to cause a fundamental paradigm shift in the way that drug discovery and development occurs.

Biomimetic artificial cells to model the effect of membrane asymmetry on chemoresistance.

We present a microfluidic platform that enables the formation of bespoke asymmetric droplet interface bilayers (DIBs) as artificial cell models from naturally-derived lipids. We use them to perform pharmacokinetic assays to quantify how lipid asymmetry affects the permeability of the chemotherapy drug doxorubicin. Previous attempts to model bilayer asymmetry with DIBs have relied on the use of synthetic lipids to achieve asymmetry. Use of natural lipids serves to increase the biomimetic nature of these artificial cells, showcasing the next step towards forming a true artificial cell membrane in vitro. Here we use our microfluidic platform to form biomimetic, asymmetric and symmetric DIBs, with their asymmetry quantified through their life-mimicking degree of curvature. We subsequently examine permeability of these membranes to doxorubicin, and reveal measurable differences in its pharmacokinetics induced by membrane asymmetry, highlighting another factor that potentially contributes to chemoresistance in some forms of cancer.

A plug-and-play modular microcapillary platform for the generation of multicompartmental double emulsions using glass or fluorocarbon capillaries.

Although multiple emulsions have a wide range of applications in biology, medicine, chemistry and cosmetics, the use of microfluidic devices to generate them remains limited to specialist laboratories. This is because of the expertise required to design and operate these technologies. Here we show a plug-and-play microcapillary platform for the generation of multicompartmental double emulsions which only requires a low cost 3D printer for fabrication and syringe pumps for operation. Our microcapillary platform is modular because we fabricate junction boxes from a flexible resin to hold and align any type of standard glass capillary or piece of tubing for droplet formation without the need for capillary alignment. The flexible resin enables total sealing of the capillaries without the need for gaskets or adhesives, and the ability to use any type of capillary or tubing means that surface treatment is not required. We show how our microcapillary platform is able to generate water-in-oil-in-water, oil-in-water-in-oil, and oil-in-oil-in-water multicompartmental double emulsions with between 1 and 10 inner droplets with high accuracy and reproducibility using standard oils (FC40, mineral oil) and inexpensive surfactants (sodium dodecyl sulfate, SDS or 1H,1H,2H,2H-perfluoro-1-octanol, PFO). Additionally, we show the formation of binary multicompartmental double emulsions, where two types of inner phase droplets can be encapsulated in the multicompartmental emulsions. Our results demonstrate how simple and accessible tools can be employed to generate a powerful modular microcapillary platform. We anticipate that the simplicity of fabrication and operation of this platform, coupled with its ability to make a wide variety of different types of emulsions, will be attractive both to microfluidic laboratories and to those without microfluidic expertise who need an enabling tool for multicompartmental double emulsion formation.

Recent advances in the design of microfluidic technologies for the manufacture of drug releasing particles.

Drug releasing particles are valued for their ability to deliver therapeutics to targeted locations and for their controllable release patterns. The development of microfluidic technologies, which are designed specifically to manipulate small amounts of fluids, to manufacture particles for drug delivery applications reflects a recent trend due to the advantages they confer in terms of control over particle size and material composition. This review takes a comprehensive look at the different types of microfluidic devices used to fabricate such particles from different types of biomaterials, and at how the on-chip features enable the production of particles with different types of properties. The review concludes by suggesting avenues for future work that will enable these technologies to fulfill their potential and be used in industrial settings for the manufacture of drug releasing particles with unique capabilities.

Programmed assembly of bespoke prototissues on a microfluidic platform.

The precise assembly of protocell building blocks into prototissues that are stable in water, capable of sensing the external environment and which display collective behaviours remains a considerable challenge in prototissue engineering. We have designed a microfluidic platform that enables us to build bespoke prototissues from predetermined compositions of two types of protein-polymer protocells. We can accurately control their size, composition and create unique Janus configurations in a way that is not possible with traditional methods. Because we can control the number and type of the protocells that compose the prototissue, we can hence modulate the collective behaviours of this biomaterial. We show control over both the amplitude of thermally induced contractions in the biomaterial and its collective endogenous biochemical reactivity. Our results show that microfluidic technologies enable a new route to the precise and high-throughput fabrication of tissue-like materials with programmable collective properties that can be tuned through careful assembly of protocell building blocks of different types. We anticipate that our bespoke prototissues will be a starting point for the development of more sophisticated artificial tissues for use in medicine, soft robotics, and environmentally beneficial bioreactor technologies.

Correction: A bespoke microfluidic pharmacokinetic compartment model for drug absorption using artificial cell membranes.

Correction for 'A bespoke microfluidic pharmacokinetic compartment model for drug absorption using artificial cell membranes' by Jaime L. Korner et al., Lab Chip, 2020, 20, 1898-1906, DOI: 10.1039/D0LC00263A.

A bespoke microfluidic pharmacokinetic compartment model for drug absorption using artificial cell membranes.

Early prediction of the rate and extent of intestinal absorption is vital for the efficient development of orally administered drugs. Here we show a new type of pharmacokinetic compartment model that shows a threefold improvement in the prediction of molecular absorption in the jejunum than the current state-of-the-art in vitro technique, parallel artificial membrane permeability assays (PAMPA). Our three-stage pharmacokinetic compartment model uses microfluidic droplets and bespoke, biomimetic artificial cells to model the path of a drug proxy from the intestinal space into the blood via an enterocyte. Each droplet models the buffer and salt composition of each pharmacokinetic compartment. The artificial cell membranes are made from the major components of human intestinal cell membranes (l-α-phosphatidylcholine, PC and l-α-phosphatidylethanolamine, PE) and sizes are comparable to human cells (∼0.5 nL). We demonstrate the use of the microfluidic platform to quantify common pharmacokinetic parameters such as half-life, flux and the apparent permeability coefficient (Papp). Our determined Papp more closely resembles that of actual intestinal tissue than PAMPA, which overestimates it by a factor of 20.

Droplet confinement and leakage: Causes, underlying effects, and amelioration strategies.

The applicability of droplet-based microfluidic systems to many research fields stems from the fact that droplets are generally considered individual and self-contained reaction vessels. This study demonstrates that, more often than not, the integrity of droplets is not complete, and depends on a range of factors including surfactant type and concentration, the micro-channel surface, droplet storage conditions, and the flow rates used to form and process droplets. Herein, a model microfluidic device is used for droplet generation and storage to allow the comparative study of forty-four different oil/surfactant conditions. Assessment of droplet stability under these conditions suggests a diversity of different droplet failure modes. These failure modes have been classified into families depending on the underlying effect, with both numerical and qualitative models being used to describe the causative effect and to provide practical solutions for droplet failure amelioration in microfluidic systems.

The past, present and potential for microfluidic reactor technology in chemical synthesis.

The past two decades have seen far-reaching progress in the development of microfluidic systems for use in the chemical and biological sciences. Here we assess the utility of microfluidic reactor technology as a tool in chemical synthesis in both academic research and industrial applications. We highlight the successes and failures of past research in the field and provide a catalogue of chemistries performed in a microfluidic reactor. We then assess the current roadblocks hindering the widespread use of microfluidic reactors from the perspectives of both synthetic chemistry and industrial application. Finally, we set out seven challenges that we hope will inspire future research in this field.

Droplet dispensing in digital microfluidic devices: Assessment of long-term reproducibility.

We report an in-depth study of the long-term reproducibility and reliability of droplet dispensing in digital microfluidic devices (DMF). This involved dispensing droplets from a reservoir, measuring the volume of both the droplet and the reservoir droplet and then returning the daughter droplet to the original reservoir. The repetition of this process over the course of several hundred iterations offers, for the first time, a long-term view of droplet dispensing in DMF devices. Results indicate that the ratio between the spacer thickness and the electrode size influences the reliability of droplet dispensing. In addition, when the separation between the plates is large, the volume of the reservoir greatly affects the reproducibility in the volume of the dispensed droplets, creating "reliability regimes." We conclude that droplet dispensing exhibits superior reliability as inter-plate device spacing is decreased, and the daughter droplet volume is most consistent when the reservoir volume matches that of the reservoir electrode.