Microfluidics in the eye: a review of glaucoma implants from an engineering perspective.

Glaucoma is a progressive optic neuropathy in the eye, which is a leading cause of irreversible blindness worldwide and currently affects over 70 million individuals. Clinically, intraocular pressure (IOP) reduction is the only proven treatment to halt the progression of glaucoma. Microfluidic devices such as glaucoma drainage devices (GDDs) and minimally invasive glaucoma surgery (MIGS) devices are routinely used by ophthalmologists to manage elevated IOP, by creating an artificial pathway for the over-accumulated aqueous humor (AH) in a glaucomatous eye, when the natural pathways are severely blocked. Herein, a detailed modelling and analysis of both the natural microfluidic pathways of the AH in the eye and artificial microfluidic pathways formed additionally by the various glaucoma implants are conducted to provide an insight into the causes of the IOP abnormality and the improvement schemes of current implant designs. The mechanisms of representative glaucoma implants have been critically reviewed from the perspective of microfluidics, and we have categorized the current implants into four groups according to the targeted drainage sites of the AH, namely Schlemm's canal, suprachoroidal space, subconjunctival space, and ocular surface. In addition, we propose to divide the development and evolution of glaucoma implant designs into three technological waves, which include microtube (1st), microvalve (2nd) and microsystem (3rd). With the emerging trends of minimal invasiveness and artificial intelligence in the development of medical implants, we envision that a comprehensive glaucoma treatment microsystem is on the horizon, which is featured with active and wireless control of IOP, real-time continuous monitoring of IOP and aqueous rate, etc. The current review could potentially cast light on the unmatched needs, challenges, and future directions of the microfluidic structural and functional designs of glaucoma implants, which would enable an enhanced safety profile, reduced complications, increased efficacy of lowering IOP and reduced IOP fluctuations, closed-loop and on-demand control of IOP, etc.

Microfluidic Printing-Based Method for the Multifactorial Study of Cell-Free Protein Networks.

Protein networks can be assembled in vitro for basic biochemistry research, drug screening, and the creation of artificial cells. Two standard methodologies are used: manual pipetting and pipetting robots. Manual pipetting has limited throughput in the number of input reagents and the combination of reagents in a single sample. While pipetting robots are evident in improving pipetting efficiency and saving hands-on time, their liquid handling volume usually ranges from a few to hundreds of microliters. Microfluidic methods have been developed to minimize the reagent consumption and speed up screening but are challenging in multifactorial protein studies due to their reliance on complex structures and labeling dyes. Here, we engineered a new impact-printing-based methodology to generate printed microdroplet arrays containing water-in-oil droplets. The printed droplet volume was linearly proportional (R2 = 0.9999) to the single droplet number, and each single droplet volume was around 59.2 nL (coefficient of variation = 93.8%). Our new methodology enables the study of protein networks in both membrane-unbound and -bound states, without and with anchor lipids DGS-NTA(Ni), respectively. The methodology is demonstrated using a subnetwork of mitogen-activated protein kinase (MAPK). It takes less than 10 min to prepare 100 different droplet-based reactions, using <1 µL reaction volume at each reaction site. We validate the kinase (ATPase) activity of MEK1 (R4F)* and ERK2 WT individually and together under different concentrations, without and with the selective membrane attachment. Our new methodology provides a reagent-saving, efficient, and flexible way for protein network research and related applications.

A One-Dollar, Disposable, Paper-Based Microfluidic Chip for Real-Time Monitoring of Sweat Rate.

Collecting sweat and monitoring its rate is important for determining body condition and further sweat analyses, as this provides vital information about physiologic status and fitness level and could become an alternative to invasive blood tests in the future. Presented here is a one-dollar, disposable, paper-based microfluidic chip for real-time monitoring of sweat rate. The chip, pasted on any part of the skin surface, consists of a skin adhesive layer, sweat-proof layer, sweat-sensing layer, and scale layer with a disk-shape from bottom to top. The sweat-sensing layer has an impressed wax micro-channel containing pre-added chromogenic agent to show displacement by sweat, and the sweat volume can be read directly by scale lines without any electronic elements. The diameter and thickness of the complete chip are 25 mm and 0.3 mm, respectively, permitting good flexibility and compactness with the skin surface. Tests of sweat flow rate monitoring on the left forearm, forehead, and nape of the neck of volunteers doing running exercise were conducted. Average sweat rate on left forearm (1156 g·m-2·h-1) was much lower than that on the forehead (1710 g·m-2·h-1) and greater than that on the nape of the neck (998 g·m-2·h-1), in good agreement with rates measured using existing common commercial sweat collectors. The chip, as a very low-cost and convenient wearable device, has wide application prospects in real-time monitoring of sweat loss by body builders, athletes, firefighters, etc., or for further sweat analyses.

High-Throughput Experimentation Using Cell-Free Protein Synthesis Systems.

Cell-free protein synthesis can enable the combinatorial screening of many different components and concentrations. However, manual pipetting methods are unfit to handle many cell-free reactions. Here, we describe a microfluidic method that can generate hundreds of unique submicroliter scale reactions. The method is coupled with a high yield cell-free system that can be applied for broad protein screening assays.

Sample-to-Answer Robotic ELISA.

Enzyme-linked immunosorbent assays (ELISA), as one of the most used immunoassays, have been conducted ubiquitously in hospitals, research laboratories, etc. However, the conventional ELISA procedure is usually laborious, occupies bulky instruments, consumes lengthy operation time, and relies considerably on the skills of technicians, and such limitations call for innovations to develop a fully automated ELISA platform. In this paper, we have presented a system incorporating a robotic-microfluidic interface (RoMI) and a modular hybrid microfluidic chip that embeds a highly sensitive nanofibrous membrane, referred to as the Robotic ELISA, to achieve human-free sample-to-answer ELISA tests in a fully programmable and automated manner. It carries out multiple bioanalytical procedures to replace the manual steps involved in classic ELISA operations, including the pneumatically driven high-precision pipetting, efficient mixing and enrichment enabled by back-and-forth flows, washing, and integrated machine vision for colorimetric readout. The Robotic ELISA platform has achieved a low limit of detection of 0.1 ng/mL in the detection of a low sample volume (15 µL) of chloramphenicol within 20 min without human intervention, which is significantly faster than that of the conventional ELISA procedure. Benefiting from its modular design and automated operations, the Robotic ELISA platform has great potential to be deployed for a broad range of detections in various resource-limited settings or high-risk environments, where human involvement needs to be minimized while the testing timeliness, consistency, and sensitivity are all desired.

Digital droplet infusion.

Infusion pumps have been widely used in clinical settings for the administration of medications and fluids. We present the digital droplet infusion (DDI) device, a low-cost, high-precision digital infusion system, utilizing a microfluidic discretization unit to convert continuous flow into precisely delivered droplet aliquots and a valving unit to control the duration and frequency of flow discretization. The DDI device relies on a distinct capillarity-dominated process of coalescence and pinch-off of droplets for flow digitization, which is monitored by a pair of conductive electrodes located before and after the junction. The digital feedback-controlled flow rate can be employed to adjust a solenoid valve for refined infusion management. With this unique digital microfluidic approach, the DDI technology enables a simple yet powerful infusion system with an ultrahigh resolution of digital droplet transfer volume, as small as 57 nL, which is three orders of magnitude lower than that of clinical standard infusion pumps, as well as a wide range of digitally adjustable infusion rates ranging from 0.1 mL h-1 to 10 mL h-1, in addition to an array of programmable infusion profiles and safety features. Its modular design enables fast assembly using only off-the-shelf and 3D-printed components. Overall, benefiting from its simple device architecture and excellent infusion performance, the DDI technology has great potential to become the next-generation clinical standard for drug delivery with its high precision and ultimate portability at a low cost.

AmbuBox: A Fast-Deployable Low-Cost Ventilator for COVID-19 Emergent Care.

We present a low-cost clinically viable ventilator design, AmbuBox, using a controllable pneumatic enclosure and standard manual resuscitators that are readily available (AmbuBag), which can be rapidly deployed during pandemic and mass-casualty events with a minimal set of components to manufacture and assemble. The AmbuBox is designed to address the existing challenges presented in the existing low-cost ventilator designs by offering an easy-to-install and simple-to-operate apparatus while maintaining a long lifespan with high-precision flow control. As an outcome, a mass-producible prototype of the AmbuBox has been devised, characterized, and validated in a bench test setup using a lung simulator. This prototype will be further investigated through clinical testing. Given the potentially urgent need for inexpensive and rapidly deployable ventilators globally, the overall design, operational principle, and device characterization of the AmbuBox system have been described in detail with open access online. Moreover, the fabrication and assembly methods have been incorporated to enable short-term producibility by a generic local manufacturing facility. In addition, a full list of all components used in the AmbuBox has been included to reflect its low-cost nature.

Digital microfluidic meter-on-chip.

The accurate monitoring and control of liquid flow at low flow rates have become increasingly important in contemporary biomedical research and industrial monitoring. Inspired by the drop-counting principle implemented in a clinical gravity drip, we propose a novel microfluidic flowmetry technology for polydimethylsiloxane (PDMS)-based conventional microfluidic devices, known as a microfluidic digital meter-on-chip (DMC), to achieve on-chip and localized microflow measurements with ultrahigh precision and a wide tunable range. The DMC technology primarily relies on capillarity, unlike a gravity drip, to induce a characteristic interfacial droplet pinch-off process, from which digital microflowmetry devices can discretize continuous flow into countable transferred liquid units with consistent quantifiable volumes. Enabled by the passive discretization principle and optical transparency, the DMC device requires no external energy input or bulky control equipment, and a non-contact wireless optical detection scheme using a smartphone can be conveniently used as a readout module. Moreover, the DMC technology achieves an ultrahigh flow-to-frequency sensitivity (6.59 Hz (µL min-1)-1) and resolution (droplet transfer volume down to 2.5 nL, nearly two orders of magnitude smaller than in previously reported work, resulting in ultralow flow rates of 1 µL min-1). In addition, the flow rate measurement range covers up to 80 µL min-1 and down to at least 150 nL min-1 (over 100 times lower than reported similar digital flowmetry on the same time scale) using the current device configuration. Benefiting from its simple device architecture and adaptability, the versatile DMC technology can be seamlessly integrated with various microfluidic and nanofluidic devices for drug delivery and biochemical analysis, serving as a promising technology platform for next-generation highly demanding microflow measurements.

Microfluidic cap-to-dispense (µCD): a universal microfluidic-robotic interface for automated pipette-free high-precision liquid handling.

Microfluidic devices have been increasingly used for low-volume liquid handling operations. However, laboratory automation of such delicate devices has lagged behind due to the lack of world-to-chip (macro-to-micro) interfaces. In this paper, we have presented the first pipette-free robotic-microfluidic interface using a microfluidic-embedded container cap, referred to as a microfluidic cap-to-dispense (µCD), to achieve a seamless integration of liquid handling and robotic automation without any traditional pipetting steps. The µCD liquid handling platform offers a generic and modular way to connect the robotic device to standard liquid containers. It utilizes the high accuracy and high flexibility of the robotic system to recognize, capture and position; and then using microfluidic adaptive printing it can achieve high-precision on-demand volume distribution. With its modular connectivity, nanoliter processability, high adaptability, and multitask capacity, µCD shows great potential as a generic robotic-microfluidic interface for complete pipette-free liquid handling automation.

Larval Zebrafish Lateral Line as a Model for Acoustic Trauma.

Excessive noise exposure damages sensory hair cells, leading to permanent hearing loss. Zebrafish are a highly tractable model that have advanced our understanding of drug-induced hair cell death, yet no comparable model exists for noise exposure research. We demonstrate the utility of zebrafish as model to increase understanding of hair cell damage from acoustic trauma and develop protective therapies. We created an acoustic trauma system using underwater cavitation to stimulate lateral line hair cells. We found that acoustic stimulation resulted in exposure time- and intensity-dependent lateral line and saccular hair cell damage that is maximal at 48-72 h post-trauma. The number of TUNEL+ lateral line hair cells increased 72 h post-exposure, whereas no increase was observed in TUNEL+ supporting cells, demonstrating that acoustic stimulation causes hair cell-specific damage. Lateral line hair cells damaged by acoustic stimulation regenerate within 3 d, consistent with prior regeneration studies utilizing ototoxic drugs. Acoustic stimulation-induced hair cell damage is attenuated by pharmacological inhibition of protein synthesis or caspase activation, suggesting a requirement for translation and activation of apoptotic signaling cascades. Surviving hair cells exposed to acoustic stimulation showed signs of synaptopathy, consistent with mammalian studies. Finally, we demonstrate the feasibility of this platform to identify compounds that prevent acoustic trauma by screening a small redox library for protective compounds. Our data suggest that acoustic stimulation results in lateral line hair cell damage consistent with acoustic trauma research in mammals, providing a highly tractable model for high-throughput genetic and drug discovery studies.

Wearable microfluidics: fabric-based digital droplet flowmetry for perspiration analysis.

The latest development in wearable technologies has attracted much attention. In particular, collection and analysis of body fluids has been a focus. In this paper, we have reported a wearable microfluidic platform made using conventional fabric materials and laser micromachining to measure the flow rate on a patterned fabric surface, referred to as digital droplet flowmetry (DDF). The proposed wearable DDF is capable of collecting and measuring continuous perspiration with high precision (96% on average) in a real-time fashion over a defined area of skin. We have introduced a theoretical model for the proposed wearable interfacial microfluidic platform, under which various design parameters have been investigated and optimized for various conditions. The novel digitalized measurement principle of DDF provides fast responses, digital readouts, system flexibility, and continuous performance of the flow measurement. Moreover, the proposed DDF platform can be conveniently implemented on regular apparel or a wearable device, and has potential to be applied to dynamic removal, collection and monitoring of biofluids for various physiological and clinical processes.

Onset of particle trapping and release via acoustic bubbles.

Trapping and sorting of micro-sized objects is one important application of lab on a chip devices, with the use of acoustic bubbles emerging as an effective, non-contact method. Acoustically actuated bubbles are known to exert a secondary radiation force (FSR) on micro-particles and stabilize them on the bubble surface, when this radiation force exceeds the external hydrodynamic forces that act to keep the particles in motion. While the theoretical expression of FSR has been derived by Nyborg decades ago, no direct experimental validation of this force has been performed, and the relationship between FSR and the bubble's ability to trap particles in a given lab on a chip device remains largely empirical. In order to quantify the connection between the bubble oscillation and the resultant FSR, we experimentally measure the amplitude of bubble oscillations that give rise to FSR and observe the trapping and release of a single microsphere in the presence of the mean flow at the corresponding acoustic parameters using an acoustofluidic device. By combining well-developed theories that connect bubble oscillations to the acoustic actuation, we derive the expression for the critical input voltage that leads to particle release into the flow, in good agreement with the experiments.