Evaluating an extraction-free sample preparation method for multiplex detection of SARS-Cov-2, influenza A/B, and RSV with implementation on a microfluidic chip.

Following the relaxation of COVID-19 restrictions, other respiratory viruses such as influenza and respiratory syncytial virus (RSV), whose transmission were decreased due to COVID-19 precautions, are rising again. Because of similar clinical features and reported co-infections, multiplex detection of SARS-CoV-2, influenza A/B, and RSV is required to use specific treatments. This study assessed an extraction-free sample preparation (heat treatment at 95°C for 3 minutes) for multiplex detection using rRT-PCR. Despite an observed Ct-delay (∆Ct) averageing 1.26 compared to the standard method, an acceptable total sensitivity of 92 % and a negative predictive value (NPV) of 96 % were obtained. Moreover, Implementation on a microfluidic chip demonstrated efficiency, maintaining an excellent correlation (R2=0.983) with the standard method. Combining this extraction-free procedure with rRT-PCR on a microfluidic chip seems promising, because it simplifies the design and reduces the cost and complexity of the integrated assay for multiplex detection of SARS-CoV-2, influenza A/B, and RSV.

Multivariate analysis of nanoparticle translocation through a nanopore to improve the accuracy of resistive pulse sensing.

The advent of nanopore-based sensors based on resistive pulse sensing gave rise to a remarkable breakthrough in the detection and characterization of nanoscale species. Some strong correlations have been reported between the resistive pulse characteristics and the particle's geometrical and physical properties. These correlations are commonly used to obtain information about the particles in commercial devices and research setups. The correlations, however, do not consider the simultaneous effect of influential factors such as particle shape and off-axis translocation, which complicates the extraction of accurate information from the resistive pulses. In this paper, we numerically studied the impact of the shape and position of particles on pulse characteristics in order to estimate the errors that arise from neglecting the influence of multiple factors on resistive pulses. We considered the sphere, oblate, and prolate particles to investigate the nanoparticle shape effect. Moreover, the trajectory dependency was examined by considering the translocation of nanoparticles away from the nanopore axis. Meanwhile, the shape effect was studied for different trajectories. We observed that the simultaneous effects of influential parameters could lead to significant errors in estimating particle properties if the coupled effects are neglected. Based on the results, we introduce the "pulse waveshape" as a novel characteristic of the resistive pulse that can be utilized as a decoupling parameter in the analysis of resistive pulses.

Development of a novel cervix-inspired tortuous microfluidic system for efficient, high-quality sperm selection.

Microfluidic systems have been extensively studied in recent years as potential alternatives for problematic conventional methods of sperm selection. However, despite the widespread use of simple straight channels in these systems, the impact of channel geometry on selected sperm quality has not been thoroughly investigated. To explore this further, we designed and fabricated serpentine microchannels with different radii of curvature, inspired by the tortuous structure of the cervix. Our results showed that in the presence of gentle backflow, microfluidic channels with a 150 µm radius of curvature significantly enhanced the quality of selected sperms when compared to straight channels. Specifically, we observed significant improvements of 7% and 9% in total motility and progressive motility, respectively, as well as 13%, 18%, and 19% improvements in VCL, VAP, and VSL, respectively. Through careful observation of the process, we discovered a unique near-wall sperm migration pattern named boundary detachment-reattachment (BDR), that was observed exclusively in curved microchannels. This pattern, which is a direct consequence of the special serpentine geometry and sperm boundary-following characteristic, contributed to the superior selection performance when combined with a fluid backflow. After determining the best channel design, we fabricated a parallelized chip consisting of 85 microchannels capable of processing 0.5 ml of raw semen within 20 minutes. This chip outperformed conventional methods of swim-up and density gradient centrifugation (DGC) in terms of motility (9% and 25% improvements, respectively), reactive oxygen species (18% and 15% improvements, respectively), and DNA fragmentation index (14% improvement to DGC). Outstanding performance and advantages such as user-friendliness, rapid selection, and independence from centrifugation make our microfluidic system a prospective sperm selection tool in clinical applications.

Experimental and Theoretical Investigation on the Dynamic Response of Ferrofluid Liquid Marbles to Steady and Pulsating Magnetic Fields.

Liquid marbles are droplets enwrapped by a layer of hydrophobic micro/nanoparticles. Due to the isolation of fluid from its environment, reduction in evaporation rate, low friction with the surfaces, and capability of manipulation even on hydrophilic surfaces, liquid marbles have attracted the attention of researchers in digital microfluidics. This study investigates the manipulation of ferrofluid liquid marbles (FLMs) under DC and pulse width-modulated (PWM) magnetic fields generated by an electromagnet for the first time. At first, the threshold of the magnetic field for manipulating these FLMs is studied. Afterward, the dynamic response of the FLMs to the DC magnetic field for different FLM volumes, coil currents, and initial distances of FLM from the coil is studied, and a theoretical model is proposed. By applying the PWM magnetic field, it is possible to gain more control over the manipulation of the FLMs on the surface and adjust their position more accurately. Results indicate that with a decrease in FLM volume, coil current, and duty cycle, the FLM step length decreases; hence, FLM manipulation is more precise. Under the PWM magnetic field, it is observed that FLM movement is not synchronous with the generated pulse, and even after the coil is turned off, FLMs keep their motion. In the end, with proper adjustment of the electromagnet pulse width, launching of FLMs at a distance farther than the coil is observed.

An integrated microfluidic device for stem cell differentiation based on cell-imprinted substrate designed for cartilage regeneration in a rabbit model.

Separating cells from the body and cultivating them in vitro will alter the function of cells. Therefore, for optimal cell culture in the laboratory, conditions similar to those of their natural growth should be provided. In previous studies, it has been shown that the use of cellular shape at the culture surface can regulate cellular function. In this work, the efficiency of the imprinting method increased by using microfluidic chip design and fabrication. In this method, first, a cell-imprinted substrate of chondrocytes was made using a microfluidic chip. Afterwards, stem cells were cultured on a cell-imprinted substrate using a second microfluidic chip aligned with the substrate. Therefore, stem cells were precisely placed on the chondrocyte patterns on the substrate and their fibroblast-like morphology was changed to chondrocyte's spherical morphology after 14-days culture in the chip without using any chemical growth factor. After chondrogenic differentiation and in vitro assessments (real-time PCR and immunocytotoxicity), differentiated stem cells were transferred on a collagen-hyaluronic acid scaffold and transplanted in articular cartilage defect of the rabbit. After 6 months, the post-transplantation analysis showed that the articular cartilage defect had been successfully regenerated in differentiated stem cell groups in comparison with the controls. In conclusion, this study showed the potency of the imprinting method for inducing chondrogenicity in stem cells, which can be used in clinical trials due to the safety of the procedure.

Effects of cone angle and length of nanopores on the resistive pulse quality.

Resistive pulse sensing (RPS) has proved to be a viable method for the detection and characterization of micro and nano particles. Modern fabrication methods have introduced different nanopore geometries for resistive pulse sensors. In this paper, we have numerically studied the effects of membrane thickness and the pore's cone angle, as the main geometrical parameters, on the sensing performance of the nanopores used for nanoparticle detection in the resistive pulse sensing method. To compare the sensing performance, three resistive pulse quality parameters were investigated - sensitivity, pulse duration and pulse amplitude. The thorough investigation on the relations between the geometrical parameters and the pulse quality parameters produced several interesting results, which were categorized and summarized for different nanopore structures (as different nanopore platforms) enabling the readers to more effectively compare them with one another. The results revealed that large cone angle and low aspect ratio nanopores have higher pulse amplitude and sensitivity, but their low duration could be a challenge in the process of detecting the resistive pulse. In addition, our results show small variation in sensitivity and duration of large cone angle nanopores with respect to pore length change, which is explained using the effective length concept and the definition of electric field strength and length. The findings of the present work can be used in practical applications where choosing the optimal pore geometry is of crucial significance. Furthermore, the results provide several possible ways to improve the resistive pulse quality for better sensing performance.

Cell-Imprinted Substrates Modulate Differentiation, Redifferentiation, and Transdifferentiation.

Differentiation of stem cells into mature cells through the use of physical approaches is of great interest. Here, we prepared smart nanoenvironments by cell-imprinted substrates based on chondrocytes, tenocytes, and semifibroblasts as templates and demonstrated their potential for differentiation, redifferentiation, and transdifferentiation. Analysis of shape and upregulation/downregulation of specific genes of stem cells, which were seeded on these cell-imprinted substrates, confirmed that imprinted substrates have the capability to induce specific shapes and molecular characteristics of the cell types that were used as templates for cell-imprinting. Interestingly, immunofluorescent staining of a specific protein in chondrocytes (i.e., collagen type II) confirmed that adipose-derived stem cells, semifibroblasts, and tenocytes can acquire the chondrocyte phenotype after a 14 day culture on chondrocyte-imprinted substrates. In summary, we propose that common polystyrene tissue culture plates can be replaced by this imprinting technique as an effective and promising way to regulate any cell phenotype in vitro with significant potential applications in regenerative medicine and cell-based therapies.

Thermal control of ionic transport and fluid flow in nanofluidic channels.

In this work, we report a nanofluidic gating mechanism that uses the thermal effect for modulating the ionic transport inside nanofluidic channels. The control of the ionic transport inside a nanochannel is demonstrated using electrical conductivity. A thermal gate controls the ionic transport more effectively than most of the other gating mechanisms previously described in the scientific literature. Gating in both bulk and overlapping electric double layer regimes can be obtained. The relatively short response time of opening and closing processes makes it a good candidate for manipulating small molecules in micro- and nanoscale devices.

Temperature sensitivity of nanochannel electrical conductance.

Electrical measurement is a widely used technique for the characterization of nanofluidic devices. The electrical conductivity of electrolytes is known to be dependent on temperature. However, the similarity of the temperature sensitivity of the electrical conductivity for bulk and nanochannels has not been validated. In this work, we present the results from experimental measurements as well as analytical modeling that show the significant difference between bulk and nanoscale. The temperature sensitivity of the electrical conductance of nanochannel is higher at low ionic concentration where the nanofluidic transport is governed by the electrostatic effects from the wall. Neglecting this effect can result in significant errors for high temperature measurements. Additionally, the temperature sensitivity of the nanochannel conductance allows to measure the enthalpy change of surface reactions at low ionic concentrations.

Microfluidic synthesis of chitosan-based nanoparticles for fuel cell applications.

A microfluidic platform is developed for the synthesis of monodisperse, 100 nm, chitosan based nanoparticles using nanogelation with ATP. The resulting nanoparticles tuned and enhanced transport and electrochemical properties of Nafion based nanocomposite membranes, which is highly favorable for fuel cell applications.