Hybrid Soft-Rigid Active Prosthetics Laboratory Exercise for Hands-On Biomechanical and Biomedical Engineering Education.

This paper introduces a hands-on laboratory exercise focused on assembling and testing a hybrid soft-rigid active finger prosthetic for biomechanical and biomedical engineering (BME) education. This hands-on laboratory activity focuses on the design of a myoelectric finger prosthesis, integrating mechanical, electrical, sensor (i.e., inertial measurement units (IMUs), electromyography (EMG)), pneumatics, and embedded software concepts. We expose students to a hybrid soft-rigid robotic system, offering a flexible, modifiable lab activity that can be tailored to instructors' needs and curriculum requirements. All necessary files are made available in an open-access format for implementation. Off-the-shelf components are all purchasable through global vendors (e.g., DigiKey Electronics, McMaster-Carr, Amazon), costing approximately USD 100 per kit, largely with reusable elements. We piloted this lab with 40 undergraduate engineering students in a neural and rehabilitation engineering upper year elective course, receiving excellent positive feedback. Rooted in real-world applications, the lab is an engaging pedagogical platform, as students are eager to learn about systems with tangible impacts. Extensions to the lab, such as follow-up clinical (e.g., prosthetist) and/or technical (e.g., user-device interface design) discussion, are a natural means to deepen and promote interdisciplinary hands-on learning experiences. In conclusion, the lab session provides an engaging journey through the lifecycle of the prosthetic finger research and design process, spanning conceptualization and creation to the final assembly and testing phases.

Investigating peak dispersion in free-flow counterflow gradient focusing due to electroosmotic flow.

Free-flow electrophoresis (FFE) has the ability to continuously separate charged solutes from complex biological mixtures. Recently, a free-flow counterflow gradient focusing mechanism has been introduced to FFE, and it offers the potential for improved resolution and versatility. However, further investigation is needed to understand the solute dispersion at the focal position. Therefore, the goal of this work is to model the impact of electroosmotic flow, which is found to produce a pressure-driven backflow to maintain the fixed counterflow inputs. Like the counterflow, this backflow has a parabolic velocity profile that must be considered when predicting the concentration distribution of a given solute. After the model is established, preliminary experimental results are presented for a qualitative comparison. Results demonstrate a reasonable agreement at low applied voltages and provide a strong framework for future experimental validation.

Investigating peak dispersion in free-flow counterflow gradient focusing.

Free-flow electrophoresis (FFE) enables the continuous separation and collection of charged solutes, and as a result, it has drawn interest as both a preparative and an analytical tool for biological applications. Recently, a free-flow counterflow gradient focusing (FF-CGF) mechanism has been proposed with the goal of improving the resolution and versatility of FFE. To realize this potential, the factors that influence solute dispersion deserve further attention, including the gradient strength and the parabolic profile of the counterflow. Therefore, the goal of this work is to develop a theoretical model to study the interplay between these factors and molecular diffusion. Overall, an asymmetric solute distribution emerges for a wide range of parameters, and this behavior can be characterized with an exponentially modified Gaussian function. Results show that FF-CGF can achieve high-resolution separations, with the potential for high-throughput protein purification. Moreover, this work provides a practical guide for optimizing experimental conditions, as well as a strong framework for understanding and developing FF-CGF further.

Microwave Heating Induced On-Demand Droplet Generation in Microfluidic Systems.

In this note, we report a simple, new method for droplet generation in microfluidic systems using integrated microwave heating. This method enables droplet generation on-demand by using microwave heating to induce Laplace pressure change at the interface of the two fluids. The distance between the interface and junction and microwave excitation power have been found to influence droplet generation. Although this method is limited in generating droplets with a high rate, the fact that it can be integrated with microwave sensing that can be used as the feedback to tune the supply flow of materials presents unique advantages for applications that require dynamic tuning of material properties in droplets.

Counterflow Gradient Focusing in Free-Flow Electrophoresis for Protein Fractionation.

With its ability to continuously separate and collect charged analytes, free-flow electrophoresis (FFE) has become a useful tool for the purification and real-time analysis of biological mixtures. This work presents a new free-flow counterflow gradient focusing (FF-CGF) mechanism that uses a novel velocity gradient to counterbalance electrophoretic migration. This counterflow gradient is created by simply introducing fluid flow through the sidewalls of the FFE chamber. The theoretical foundation and device design for FF-CGF are provided in this work, followed by implementation and validation, with a detailed discussion on future opportunities and challenges. Initial results show promise, with the potential to improve FFE resolution and offer versatility. Compared to existing focusing techniques, such as free-flow isotachophoresis and isoelectric focusing, no complex buffer compositions are required.

Counterflow gradient electrophoresis for focusing and elution.

Counterflow gradient electrofocusing uses the bulk flow of a liquid solution to counterbalance the electrophoretic migration of an analyte. When either the bulk velocity or the electrophoretic velocity of the analyte is made to vary across the length of the channel, there exists a unique zero-velocity point for the analyte. This focusing method enables simultaneous separation and concentration of different analytes. The high resolution and sensitivity achieved are similar to that of isoelectric focusing, which separates analytes based on their isoelectric points, but the key difference is that analytes will instead focus based on their electrophoretic mobility. Dynamically changing the applied voltage or the counterflow rate over time will shift the zero-velocity point, and therefore allows the focused analytes to pass through a fixed detection point, or elute from the separation channel. Throughout the review, a number of different counterflow gradient techniques will be discussed, along with their recent advancements and potential applications.

Effective Thermo-Capillary Mixing in Droplet Microfluidics Integrated with a Microwave Heater.

In this study, we present a microwave-based microfluidic mixer that allows rapid mixing within individual droplets efficiently. The designed microwave mixer is a coplanar design with a small footprint, which is fabricated on a glass substrate and integrated with a microfluidic chip. The mixer works essentially as a resonator that accumulates an intensive electromagnetic field into a spiral capacitive gap (around 200 µm), which provides sufficient energy to heat-up droplets that pass through the capacitive gap. This microwave actuation induces nonuniform Marangoni stresses on the interface, which results in three-dimensional motion inside the droplet and thus fast mixing. In order to evaluate the performance of the microwave mixer, droplets with highly viscous fluid, 75% (w/w) glycerol solution, were generated, half of which were seeded with fluorescent dye for imaging purposes. The relative importance of different driving forces for mixing was evaluated qualitatively using magnitude analysis, and the effect of the applied power on mixing performance was also investigated. Mixing efficiency was quantified using the mixing index, which shows as high as 97% mixing efficiency was achieved within the range of milliseconds. This work demonstrates a very unique approach of utilizing microwave technology to facilitate mixing in droplet microfluidics systems, which can potentially open up areas for biochemical synthesis applications.

Droplet Microfluidic System with On-Demand Trapping and Releasing of Droplet for Drug Screening Applications.

96-Well plate has been the traditional method used for screening drug compounds libraries for potential bioactivity. Although this method has been proven successful in testing dose-response analysis, the microliter consumption of expensive reagents and hours of reaction and analysis time call for innovative methods for improvements. This work demonstrates a droplet microfluidic platform that has the potential to significantly reduce the reagent consumption and shorten the reaction and analysis time by utilizing nanoliter-sized droplets as a replacement of wells. This platform is evaluated by applying it to screen drug compounds that inhibit the tau-peptide aggregation, a phenomena related to Alzheimer's disease. In this platform, sample reagents are first dispersed into nanolitre-sized droplets by an immiscible carrier oil and then these droplets are trapped on-demand in the downstream of the microfluidic device. The relative decrease in fluorescence through drug inhibition is characterized using an inverted epifluorescence microscope. Finally, the trapped droplets are released on-demand after each test by manipulating the applied pressures to the channel network which allows continuous processing. The testing results agree well with that obtained from 96-well plates with much lower sample consumption (∼200 times lower than 96-well plate) and reduced reaction time due to increased surface volume ratio (2.5 min vs 2 h).

Concurrent Modeling of Hydrodynamics and Interaction Forces Improves Particle Deposition Predictions.

It is widely believed that media surface roughness enhances particle deposition-numerous, but inconsistent, examples of this effect have been reported. Here, a new mathematical framework describing the effects of hydrodynamics and interaction forces on particle deposition on rough spherical collectors in absence of an energy barrier was developed and validated. In addition to quantifying DLVO force, the model includes improved descriptions of flow field profiles and hydrodynamic retardation functions. This work demonstrates that hydrodynamic effects can significantly alter particle deposition relative to expectations when only the DLVO force is considered. Moreover, the combined effects of hydrodynamics and interaction forces on particle deposition on rough, spherical media are not additive, but synergistic. Notably, the developed model's particle deposition predictions are in closer agreement with experimental observations than those from current models, demonstrating the importance of inclusion of roughness impacts in particle deposition description/simulation. Consideration of hydrodynamic contributions to particle deposition may help to explain discrepancies between model-based expectations and experimental outcomes and improve descriptions of particle deposition during physicochemical filtration in systems with nonsmooth collector surfaces.

Microfluidic two-dimensional separation of proteins combining temperature gradient focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

A two-dimensional separation system is presented combining scanning temperature gradient focusing (TGF) and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) in a PDMS/glass microfluidic chip. Denatured proteins are first focused and separated in a 15 mm long channel via TGF with a temperature range of 16-47 °C and a pressure scanning rate of -0.5 Pa/s and then further separated via SDS-PAGE in a 25 mm long channel. A side channel is designed at the intersection between the two dimensions to continuously inject SDS into the gel, allowing SDS molecules to be compiled within the focused bands. Separation experiments are performed using several fluorescently labeled proteins with single point detection. Experimental results show a dramatic improvement in peak capacity over one-dimensional separation techniques.

Model of separation performance of bilinear gradients in scanning format counter-flow gradient electrofocusing techniques.

Counter-flow gradient electrofocusing allows the simultaneous concentration and separation of analytes by generating a gradient in the total velocity of each analyte that is the sum of its electrophoretic velocity and the bulk counter-flow velocity. In the scanning format, the bulk counter-flow velocity is varying with time so that a number of analytes with large differences in electrophoretic mobility can be sequentially focused and passed by a single detection point. Studies have shown that nonlinear (such as a bilinear) velocity gradients along the separation channel can improve both peak capacity and separation resolution simultaneously, which cannot be realized by using a single linear gradient. Developing an effective separation system based on the scanning counter-flow nonlinear gradient electrofocusing technique usually requires extensive experimental and numerical efforts, which can be reduced significantly with the help of analytical models for design optimization and guiding experimental studies. Therefore, this study focuses on developing an analytical model to evaluate the separation performance of scanning counter-flow bilinear gradient electrofocusing methods. In particular, this model allows a bilinear gradient and a scanning rate to be optimized for the desired separation performance. The results based on this model indicate that any bilinear gradient provides a higher separation resolution (up to 100%) compared to the linear case. This model is validated by numerical studies.

Microchip with an open tubular immobilized ph gradient for UV whole column imaging detection.

This study reports a new method for establishing an open tubular IPG in a microchip coupled with a whole column image detection (WCID) system for protein separation applications. This method allows a wider range of immobilized pH (2.6-9.5) to be established in a PDMS/quartz channel by controlling the diffusion of acidic and basic polymer solutions into the channel through well-designed channel dimensions. The developed pH gradient was experimentally validated by performing the separation of a mixture of standard pI markers. It was further validated by the separation of the hemoglobin control AFSC sample. This method is advantageous over existing IPG methods because it has a wider range of pH and maintains the open tubular feature that matches the UV WCID to improve the sensitivity.

Soft robotics-inspired sensing system for detecting downward movement and pistoning in prosthetic sockets: A proof-of-concept study.

BACKGROUND: Vertical displacement of the residual limb within transtibial prosthetic socket, often known as "pistoning" or downward movement, may lead to skin breakdowns and ulcers. Downward movement is particularly difficult to self-manage for diabetic individuals living with amputation because of diminished sensation in the residual limb from peripheral neuropathy. Therefore, a customizable sensor at the distal end that can alert the users when high-risk downward movement and pistoning occurs is urgently needed. OBJECTIVES: Presented herein for the first time is a lightweight, inexpensive sensing system inspired by soft robotics that can detect the occurrence and severity of downward movement at the distal end. METHODS: The sensing system consists of a multilayered torus-shaped balloon, allowing easy integration with pin-lock socket systems. The design allows sensing of vertical displacement without imparting high reaction forces back onto the distal end. A benchtop compression tester was used to characterize system performance. Systematic and parametric benchtop tests were conducted to examine the sensor's physical characteristics. Long-term (24-h) stability of the sensor was also recorded. RESULTS: Compared with water, air was determined to be a better medium with a higher linear full-scale span (FSS) because of its compressible nature. Repeatable 0.5-mm vertical displacements yielded a linear (>0.99 R2) FSS of 4.5 mm and a sensitivity of 0.8 kPa/mm. The sensing system is highly precise, with as low as 1% FSS total error band and average hysteresis of 2.84% of FSS. Over 24 h, a 4% FSS drift was observed. CONCLUSION: Sensing system characteristics, coupled with low-cost, customizable fabrication, indicates promising performance for daily use to notify and alert transtibial prosthetic users of downward movement and/or pistoning.

Bilinear temperature gradient focusing in a hybrid PDMS/glass microfluidic chip integrated with planar heaters for generating temperature gradients.

Temperature gradient focusing (TGF) is a counterflow gradient focusing technique, which utilizes a temperature gradient across a microchannel or capillary to separate analytes. With an appropriate buffer, the temperature gradient creates a gradient in both the electric field and electrophoretic velocity. Combined with a bulk counter flow, ionic species concentrate at a unique point where the total velocity sums to zero and separate from each other. Scanning TGF uses varying bulk flow so that a large number of analytes that have large differences in electrophoretic mobility can be sequentially focused and passed by a single detection point. Up to now, scanning TGF examples have been performed using a linear temperature gradient which has limitations in improving peak capacity and resolution at the same time. In this work, we develop a bilinear temperature gradient along the separation channel that improves both peak capacity and separation resolution simultaneously. The temperature profile along the channel consists of a very sharp gradient used to preconcentrate the sample followed by a shallow gradient that increases separation resolution. A specialized design is developed for the heaters to achieve the bilinear profile using both analytical and numerical modeling. The heaters are integrated onto a hybrid PDMS/glass chip fabricated using conventional sputtering and soft-lithography techniques. Separation performance is characterized by separating several different dyes and amino acids that have close electrophoretic mobilities. Experiments show a dramatic improvement in peak capacity and resolution in comparison to the standard linear temperature gradient.

Micellar affinity gradient focusing in a microfluidic chip with integrated bilinear temperature gradients.

Micellar affinity gradient focusing (MAGF) is a microfluidic counterflow gradient focusing technique that combines the favorable features of MEKC and temperature gradient focusing. MAGF separates analytes on the basis of a combination of electrophoretic mobility and partitioning with the micellar phase. A temperature gradient is produced along the separation channel containing an analyte/micellar system to create a gradient in interaction strength (retention factor) between the analytes and micelles. Combined with a bulk counterflow, species concentrate at a unique point where their total velocity sums to zero. MAGF can be used in scanning mode by varying the bulk flow so that a large number of analytes can be sequentially focused and passed by a single detection point. In this work, we develop a bilinear temperature gradient along the separation channel that improves separation performance over the conventional linear designs. The temperature profile along the channel consists of a very sharp gradient used to preconcentrate the sample followed by a shallow gradient that increases resolution. We fabricated a hybrid PDMS/glass microfluidic chip with integrated micro heaters that generate the bilinear profile. Performance is characterized by separating several different samples including fluorescent dyes using SDS surfactant and pI markers using both SDS and poly-SUS surfactants as the micellar phase. The new design shows a nearly two times improvement in peak capacity and resolution in comparison to the standard linear temperature gradient.

A new soft lithographic route for the facile fabrication of hydrophilic sandwich microchips.

Manufacturing materials are an essential element for the fabrication of microfluidic chips. PDMS, the most widely used polymeric material, is associated with apparent disadvantages such as hydrophobic nature, while other materials also suffer from some limitations. In this paper, a new soft lithographic route was proposed for the facile manufacturing of hydrophilic sandwich microchips, using bisphenol A based epoxy acrylate (BABEA) as a new patterning material. The BABEA copolymers are hydrophilic, highly transparent in visible range while highly untransparent when the wavelength is less than 290 nm, and of high replication fidelity. By combining with appropriate monomers, including glycidyl methacrylate, methylmethacrylate, and acrylic acid, the copolymers contain active functional groups, which allows for easy postmodification for desirable functional units. A fabrication procedure was proposed for manufacturing hybrid quartz/BABEA copolymer/quartz microchips. In the procedure, no micromachining equipments, wet etching, or imprinting techniques were involved, making the fabrication approach applicable in ordinary chemistry laboratories. The performance of the prepared microchips was demonstrated in terms of CIEF with UV-whole channel imaging detection. The hydrophilic microchannel ensures stable focusing while the polymeric middle layer acts as a perfectly aligned optical slit for whole channel UV absorbance detection.

Droplet formation of biological non-Newtonian fluid in T-junction generators. I. Experimental investigation.

The extension of microfluidics to many bioassay applications requires the ability to work with non-Newtonian fluids. One case in point is the use of microfluidics with blood having different hematocrit levels. This work is the first part of a two-part study and presents the formation dynamics of blood droplets in a T-junction generator under the squeezing regime. In this regime, droplet formation with Newtonian fluids depends on T-junction geometry; however, we found that in the presence of the non-Newtonian fluid such as red blood cells, the formation depends on not only to the channel geometry, but also the flow rate ratio of fluids, and the viscosity of the phases. In addition, we analyzed the impact of the red blood cell concentration on the formation cycle. In this study, we presented the experimental data of the blood droplet evolution through the analysis of videos that are captured by a high-speed camera. During this analysis, we tracked several parameters such as droplet volume, spacing between droplets, droplet generation frequency, flow conditions, and geometrical designs of the T junction. Our analysis revealed that, unlike other non-Newtonian fluids, where the fourth stage exists (stretching stage), the formation cycle consists of only three stages: lag, filling, and necking stages. Because of the detailed analysis of each stage, a mathematical model can be generated to predict the final volume of the blood droplet and can be utilized as a guide in the operation of the microfluidic device for biochemical assay applications; this is the focus of the second part of this study [Phys. Rev. E 105, 025106 (2022)10.1103/PhysRevE.105.025106].

Droplet formation of biological non-Newtonian fluid in T-junction generators. II. Model for final droplet volume prediction.

This work represents the second part of a two-part series on the dynamics of droplet formation in a T-junction generator under the squeezing regime when using solutions of red blood cells as the dispersed phase. Solutions containing red blood cells are non-Newtonian; however, these solutions do not behave in the same way as other non-Newtonian fluids currently described in the literature. Hence, available models do not capture nor predict important features useful for the design of T-junction microfluidic systems, including droplet volume. The formation of a red blood cell-containing droplet consists of three stages: a lag stage, a filling stage, and a necking stage, with the lag stage only observed in narrow dispersed phase channel setups. Unlike other shear-thinning fluids, thread elongation into the main channel at the end of the necking stage is not observed for red blood cell solutions. In this work, a model that predicts the final droplet volume of a red blood cell containing droplets in T-junction generators is presented. The model combines a detailed analysis of the geometrical shape of the droplet during the formation process, with force and Laplace pressure balances to obtain the penetration depth (b_{fill}^{*}) and the critical neck thickness (2r_{pinch}^{*}) of the droplet. The performance of the model was validated by comparing the operational parameters (droplet volume, the spacing between the droplet, and the generation frequency) with the experimental data across a range of the dimensionless parameters (flow rate ratios, continuous phase viscosities, and channel geometries).

Reagent free detection of SARS-CoV-2 using an antibody-based microwave sensor in a microfluidic platform.

The global COVID-19 pandemic caused by SARS-CoV-2 has resulted in an unprecedented economic and societal impact. Developing simple and accurate testing methods for point-of-care (POC) diagnosis is crucial not only for the control of COVID-19, but also for better response to similar outbreaks in the future. In this work, we present a novel proof-of-concept of a microfluidic microwave sensing method for POC diagnosis of the SARS-CoV-2 virus. This method relies on the antibody immobilized on the microwave sensor to selectively capture and concentrate the SARS-CoV-2 antigen or virus present in a buffer solution flowing through the sensor region in a microchannel. The capturing of the SARS-CoV-2 antigen or virus results in a change in the permittivity of the medium near the sensor region reflected by the resonance frequency shift which is used for detection. The use of microchannels offers precise control of the sample volume and the continuous flow nature also offers the potential to monitor the dynamic capturing process. The microwave-microfluidic device shows a good sensitivity of 0.1 ng ml-1 for the SARS-CoV-2 antigen and 4000 copies per ml for the SARS-CoV-2 virus. The resonance frequency shift presents a linear relationship with the logarithm of antigen or virus concentration, respectively. This detection method is able to distinguish SARS-CoV-2 from the antigen of human CD4 and two human coronaviruses (MERS and HKU1), which presents a new pathway towards POC diagnosis of the COVID-19 at the community level. It presents the potential to detect other viruses by functionalizing the microwave sensor with respective antibodies.

Shear-Thinning and Temperature-Dependent Viscosity Relationships of Contemporary Ocular Lubricants.

PURPOSE: To evaluate the shear viscosity of contemporary, commercially available ocular lubricants at various shear rates and temperatures and to derive relevant mathematical viscosity models that are impactful for prescribing and developing eye drops to treat dry eye disease. METHODS: The shear viscosity of 12 ocular lubricants was measured using a rheometer and a temperature-controlled bath at clinically relevant temperatures at which users may experience exposure to the drops (out of the refrigerator [4.3°C]; room temperature [24.6°C]; ocular surface temperature [34.5°C]). Three replicates for each sample at each temperature were obtained using a standard volume (0.5 mL) of each sample. The viscosity of each ocular lubricant was measured over the full range of shear rates allowed by the rheometer. RESULTS: The shear viscosity of the same ocular lubricant varied significantly among the three temperatures. In general, a higher temperature resulted in smaller viscosities than a lower temperature (an average of -48% relative change from 4.3°C to 24.6°C and -21% from 24.6°C to 34.5°C). At a constant temperature, the viscosity of an ocular lubricant over the studied shear rates can be well approximated by a power-law model. CONCLUSIONS: Rheological analysis revealed that the ocular lubricants exhibited shear-thinning behavior at the measured temperatures. Differences in the ocular lubricants' formulations and measured temperatures resulted in different viscosities. TRANSLATIONAL RELEVANCE: When prescribing eye drops, eye care professionals can select the optimal one for their patients by considering a variety of factors, including its rheological property at physiologically relevant shear rates and temperatures, which can improve residence time on the ocular surface, while ensuring appropriate comfort and vision. However, care must be taken when using the derived mathematical models in this study because the in vivo shear behavior of the ocular lubricants has not been examined and might show deviations from those reported when placed on the ocular surface.