Detection of Interleukin-1 ß (IL-1ß) in Single Human Blastocyst-Conditioned Medium Using Ultrasensitive Bead-Based Digital Microfluidic Chip and Its Relationship with Embryonic Implantation Potential.

The implantation of human embryos is a complex process involving various cytokines and receptors expressed by both endometrium and embryos. However, the role of cytokines produced by a single embryo in successful implantation is largely unknown. This study aimed to investigate the role of IL-1ß expressed in a single-embryo-conditioned medium (ECM) in embryo implantation. Seventy samples of single ECM were analyzed by a specially designed magnetic-beads-based microfluidic chip from 15 women. We discovered that IL-1ß level increased as the embryo developed, and the difference was significant. In addition, receiver operator characteristic (ROC) curves analysis showed a higher chance of pregnancy when the IL-1ß level on day 5 ECM was below 79.37 pg/mL and the difference between day 5 and day 3 was below 24.90 pg/mL. Our study discovered a possible association between embryonic proteomic expression and successful implantation, which might facilitate single-embryo transfer in the future by helping clinicians identify the embryo with the greatest implantation potential.

Microfluidic Microalgae System: A Review.

Microalgae that have recently captivated interest worldwide are a great source of renewable, sustainable and economical biofuels. The extensive potential application in the renewable energy, biopharmaceutical and nutraceutical industries have made them necessary resources for green energy. Microalgae can substitute liquid fossil fuels based on cost, renewability and environmental concern. Microfluidic-based systems outperform their competitors by executing many functions, such as sorting and analysing small volumes of samples (nanolitre to picolitre) with better sensitivities. In this review, we consider the developing uses of microfluidic technology on microalgal processes such as cell sorting, cultivation, harvesting and applications in biofuels and biosensing.

A Review on Microfluidics: An Aid to Assisted Reproductive Technology.

Infertility is a state of the male or female reproductive system that is defined as the failure to achieve pregnancy even after 12 or more months of regular unprotected sexual intercourse. Assisted reproductive technology (ART) plays a crucial role in addressing infertility. Various ART are now available for infertile couples. Fertilization in vitro (IVF), intracytoplasmic sperm injection (ICSI) and intrauterine insemination (IUI) are the most common techniques in this regard. Various microfluidic technologies can incorporate various ART procedures such as embryo and gamete (sperm and oocyte) analysis, sorting, manipulation, culture and monitoring. Hence, this review intends to summarize the current knowledge about the application of this approach towards cell biology to enhance ART.

Synthesis and characterization of magnetic nanoparticles coated with polystyrene sulfonic acid for biomedical applications.

The development of novel magnetic nanoparticles (MNPs) with satisfactory biocompatibility for biomedical applications has been the subject of extensive exploration over the past two decades. In this work, we synthesized superparamagnetic iron oxide MNPs coated with polystyrene sulfonic acid (PSS-MNPs) and with a conventional co-precipitation method. The core size and hydrodynamic diameter of the PSS-MNPs were determined as 8-18 nm and 50-200 nm with a transmission electron microscopy and dynamic light scattering, respectively. The saturation magnetization of the particles was measured as 60 emu g-1 with a superconducting quantum-interference-device magnetometer. The PSS content in the PSS-MNPs was 17% of the entire PSS-MNPs according to thermogravimetric analysis. Fourier-transform infrared spectra were recorded to detect the presence of SO3 - groups, which confirmed a successful PSS coating. The structural properties of the PSS-MNPs, including the crystalline lattice, composition and phases, were characterized with an X-ray powder diffractometer and 3D nanometer-scale Raman microspectrometer. MTT assay and Prussian-blue staining showed that, although PSS-MNPs caused no cytotoxicity in both NIH-3T3 mouse fibroblasts and SK-HEP1 human liver-cancer cells up to 1000 µg mL-1, SK-HEP1 cells exhibited significantly greater uptake of PSS-MNPs than NIH-3T3 cells. The low cytotoxicity and high biocompatibility of PSS-MNPs in human cancer cells demonstrated in the present work might have prospective applications for drug delivery.

Get to Understand More from Single-Cells: Current Studies of Microfluidic-Based Techniques for Single-Cell Analysis.

This review describes the microfluidic techniques developed for the analysis of a single cell. The characteristics of microfluidic (e.g., little sample amount required, high-throughput performance) make this tool suitable to answer and to solve biological questions of interest about a single cell. This review aims to introduce microfluidic related techniques for the isolation, trapping and manipulation of a single cell. The major approaches for detection in single-cell analysis are introduced; the applications of single-cell analysis are then summarized. The review concludes with discussions of the future directions and opportunities of microfluidic systems applied in analysis of a single cell.

Investigation of C-terminal domain of SARS nucleocapsid protein-Duplex DNA interaction using transistors and binding-site models.

AlGaN/GaN high electron mobility transistors (HEMTs) were used to sense the binding between double stranded DNA (dsDNA) and the severe acute respiratory syndrome coronavirus (SARS-CoV) nucleocapsid protein (N protein). The sensing signals were the drain current change of the HEMTs induced by the protein-dsDNA binding. Binding-site models using surface coverage ratios were utilized to analyze the signals from the HEMT-based sensors to extract the dissociation constants and predict the number of binding sites. Two dissociation constants, K D1 = 0.0955 nM, K D2 = 51.23 nM, were obtained by fitting the experimental results into the two-binding-site model. The result shows that this technique is more competitive than isotope-labeling electrophoretic mobility shift assay (EMSA). We demonstrated that AlGaN/GaN HEMTs were highly potential in constructing a semiconductor-based-sensor binding assay to extract the dissociation constants of nucleotide-protein interaction.

A Novel EWOD Platform for Freely Transporting Droplets in Double and Single-Plate Structures.

This study developed a novel dielectric wetting microfluidic operation platform combining parallel-plate and coplanar-plate regions with a curved surface structure as the connection structure. With the new electrowetting on dielectric (EWOD) platform, "droplet pull-out" has been successfully achieved and viewed as an essential new operation for microfluidics with the dielectric wetting technique. The EWOD system is divided into a PDMS substrate top plate and an indium tin oxide (ITO) glass substrate as a bottom layer on this chip. In the parallel-plate region, the droplets can be generated and transported through the square parallel electrodes; in the single-plate area, the droplets can be pulled out from the parallel structure, transported and mixed through the common grounded coplanar electrodes. In dielectric wetting performance testing, coplanar electrodes can apply a maximum driving force of 31.22 µN to DI water and 13.38 µN to propylene carbonate (PC). This driving force is sufficient to detach the sample from the top cover and pull the sub-droplet from the parallel plate structure for DI water, PC and polyethylene glycol diacrylate (PEGDA) buffer. The novel EWOD system also possesses the advantage of precise volume control for liquid samples; the volume error of the generated droplet can be controlled within 0.1% to 2%.

Pioneering Data Processing for Convolutional Neural Networks to Enhance the Diagnostic Accuracy of Traditional Chinese Medicine Pulse Diagnosis for Diabetes.

Traditional Chinese medicine (TCM) has relied on pulse diagnosis as a cornerstone of healthcare assessment for thousands of years. Despite its long history and widespread use, TCM pulse diagnosis has faced challenges in terms of diagnostic accuracy and consistency due to its dependence on subjective interpretation and theoretical analysis. This study introduces an approach to enhance the accuracy of TCM pulse diagnosis for diabetes by leveraging the power of deep learning algorithms, specifically LeNet and ResNet models, for pulse waveform analysis. LeNet and ResNet models were applied to analyze TCM pulse waveforms using a diverse dataset comprising both healthy individuals and patients with diabetes. The integration of these advanced algorithms with modern TCM pulse measurement instruments shows great promise in reducing practitioner-dependent variability and improving the reliability of diagnoses. This research bridges the gap between ancient wisdom and cutting-edge technology in healthcare. LeNet-F, incorporating special feature extraction of a pulse based on TMC, showed improved training and test accuracies (73% and 67%, respectively, compared with LeNet's 70% and 65%). Moreover, ResNet models consistently outperformed LeNet, with ResNet18-F achieving the highest accuracy (82%) in training and 74% in testing. The advanced preprocessing techniques and additional features contribute significantly to ResNet18-F's superior performance, indicating the importance of feature engineering strategies. Furthermore, the study identifies potential avenues for future research, including optimizing preprocessing techniques to handle pulse waveform variations and noise levels, integrating additional time-frequency domain features, developing domain-specific feature selection algorithms, and expanding the scope to other diseases. These advancements aim to refine traditional Chinese medicine pulse diagnosis, enhancing its accuracy and reliability while integrating it into modern technology for more effective healthcare approaches.

A flexible hydrophilic-modified graphene microprobe for neural and cardiac recording.

A graphene-based flexible microprobe developed by microelectromechanical system technology shows high resolution for the detection of electrophysiological signals from various bio-objects. The hydrophilization post-treatment using steam plasma was performed on the graphene surface to decrease the interfacial impedance between graphene and electrolyte, and thus improve the signal-to-noise ratio during neural and cardiac recording. The signal-to-noise ratio of the action potentials from axons of a crayfish measured by hydrophilic-modified graphene microprobe (27.8±4.0dB) is higher than that of untreated device (20.3±3.3dB). Also, the form of the QRS complex and T wave in the electrocardiogram of the zebrafish heart can be clearly distinguished using the modified device. The total measured noise levels of the overall stability of the system were 4.2µVrms (hydrophilic graphene) and 7.64µVrms (hydrophobic graphene). The graphene-based implant can be further used for in vivo, long-term recording and retina prosthesis. FROM THE CLINICAL EDITOR: In this study a graphene-based flexible microprobe developed using microelectromechanical system technology was demonstrated to enable high resolution detection of electrophysiological signals, including EKG in zebrafish models. Both hydrophilic and hydrophobic graphene were studied, paving the way to potential future clinical applications of this new technology.

Programmable UV-Curable Resin by Dielectric Force.

In this study, UV-curable resin was formed into different patterns through the programmable control of dielectric force. The dielectric force is mainly generated by the dielectric chip formed by the interdigitated electrodes. This study observed that of the control factors affecting the size of the UV resin driving area, current played an important role. We maintained the same voltage-controlled condition, changing the current from 0.1 A to 0.5 A as 0.1 A intervals. The area of droplets was significantly different at each current condition. On the other hand, we maintained the same current condition, and changed the voltage from 100 V to 300 V at 50 V intervals. The area of droplets for each voltage condition was not obviously different. The applied frequency of the AC (Alternating Current) electric field increased from 10 kHz to 50 kHz. After driving the UV resin, the pattern line width of the UV resin could be finely controlled from 224 um to 137 um. In order to form a specific pattern, controlling the current and frequency could achieved a more accurate shape. In this article, UV resin with different patterns was formed through the action of this dielectric force, and after UV curing, tiny structural parts could be successfully demonstrated.

Polymer/Ordered mesoporous carbon nanocomposite platelets as superior sensing materials for gas detection with surface acoustic wave devices.

We have prepared nanocomposites of polymers and platelet CMK-5-like carbon and have demonstrated their superior performance for gravimetric gas detection. The zirconium-containing platelet SBA-15 was used as hard template to prepare CMK-5-like carbon, which was then applied as a lightweight and high-surface-area scaffold for the growth of polymers by radical polymerization. Mesoporous nanocomposites composed of four different polymers were used as sensing materials for surface acoustic wave devices to detect ppm-level ammonia gas. The sensors showed much better sensitivity and reversibility than those coated with dense polymer films, and the sensor array could still generate a characteristic pattern for the analyte with a concentration of 16 ppm. The results show that the nanocomposite sensing materials are promising for highly sensitive gravimetric-type electronic nose applications.

Contactless Micro-Droplet Manipulation of Liquid Released from a Parallel Plate to an Open Region in Electrowetting-on-Dielectric Platform.

In electrowetting-on-dielectric (EWOD) platform, the transfer of droplets from the EWOD boundary region (top plate and bottom plate) to the open region is challenging. The challenge is due to the resistance-like surface tension, friction from the top-plate edge, and the so-called boundary. For this purpose, we designed the top plate to minimize the friction resistance at the boundary. The experiment focused on Gibb's formula and successfully transferred the liquid droplet between the top plate and bottom plate boundary region under a high voltage environment. The threshold voltage for the successful transportation of the droplet between the boundary is 250 V which provides strong pressure to drive the droplet.

Identification of Microorganisms Using an EWOD System.

Among the advantages of an electrowetting-on-dielectric (EWOD) chip are its uncomplicated fabrication and low cost; one of its greatest strengths that might be applied in the field of biomedical technology is that it can accurately control volume and reduces the amount of samples and reagents. We present an EWOD for the biochemical identification of microorganisms, which is required to confirm the source of microbial contamination or quality inspection of product-added bacteria, etc. The traditional kit we used existed in the market; the detection results are judged by the pattern of color change after incubation. After a preliminary study, we confirmed that an image-processing tool (ImageJ) provides a suitable method of analysis, and that, when the concentration of the sugar reagent is 38 µg/µL, the best operating parameters for the EWOD chip in silicone oil are 40 V and 1.5 kHz. Additionally, we completed the biochemical identification of five bacterial species on the EWOD chip at the required concentration of the kit. Next, we found a decreased duration of reaction and that the least number of bacteria that were identifiable on the chip lies between 100 and 1000 CFU per droplet. Because the number of bacteria required on the chip is much smaller than for the kit, we tested whether a single colony can be used for identification, which provided a positive result. Finally, we designed an experimental flow to simulate an actual sample in an unclean environment, in which we divided the various processed samples into four groups to conduct experiments on the chip.

Detection of the Freshness of Kiwifruit With a TD-GC-MS and a Gas-Sensing Array Based on the Surface-Acoustic-Wave Technique.

An electronic nose is an arrayed gas sensor mimicking the human olfactory system that can analyze and identify a flavour on collecting an odour from an environment. In our experiments, an electronic-nose system based on a surface acoustic wave (SAW) was used to measure the freshness of kiwifruit. 128° YX-LiNbO3 acted as a piezoelectric material; Au was deposited as an electrode and sensing area. With a polymer coating of various types on the sensing area and a connection to an oscillator circuit, a 113-114 MHz SAW was obtained. Depending on the properties of varied polymers, the frequency shift varied due to absorbed volatile organic compounds (VOC). In this way, with four surface-acoustic-wave sensors coated with varied polymers we built a kiwi-flavour database according to results from a TD-GC-MS system. When the concentration of esters increased, the kiwifruit began to ripen, accompanied by increased concentrations and types of VOC. As a result, polystyrene (PS) and fluoropolymer (CYTOP) polymers, which played the role of sensing materials, served as major materials to determine the ester aroma profile. Polyvinyl alcohol (PVA), polyvinyl butyral (PVB) and poly-N-vinylpyrrolidone (PNVP) were used to trap the alcohols and acids during a kiwifruit ripening period. This research proved that discrimination of differences is feasible from an unripe stage to a ripe stage and from a ripe stage to an over-ripe stage.

A High-Voltage TENG-Based Droplet Energy Generator With Ultralow Liquid Consumption.

A solid-liquid triboelectric nanogenerator (TENG) has attracted increasing research interest in relation to the development of regeneration energy based on water resources. The output of solid-liquid TENG remains unsolved, however, because of the low voltage output that impedes wide applications. To this end, in this work we developed a miniaturized microfluidic channel-based TENG device for highly efficient conversion of energy from the transport of a water droplet to an electrical output. We investigated an optimized design in a triboelectric material, the droplet transport and the electrostatic induction layer to provide a high voltage output and stable energy harvesting. The optimized device demonstrated maximum voltage amplitude 102 mV with an ultralow liquid consumption, 0.36 [Formula: see text], resulting in sample-energy conversion 283.33 mV/ [Formula: see text]. This novel device is expected potentially to address the limitations imposed by sample consumption in energy harvesting in the future.

Sensitivity Enhancement and Probiotic Detection of Microfluidic Chips Based on Terahertz Radiation Combined with Metamaterial Technology.

Terahertz (THz) radiation has attracted wide attention in recent years due to its non-destructive properties and ability to sense molecular structures. In applications combining terahertz radiation with metamaterial technology, the interaction between the terahertz radiation and the metamaterials causes resonance reactions; different analytes have different resonance performances in the frequency domain. In addition, a microfluidic system is able to provide low volume reagents for detection, reduce noise from the environment, and concentrate the sample on the detection area. Through simulation, a cruciform metamaterial pattern was designed; the proportion, periodicity, and width of the metamaterial were adjusted to improve the sensing capability of the chip. In the experiments, the sensing capabilities of Type A, B, and C chips were compared. The Type C chip had the most significant resonant effect; its maximum shift could be increased to 89 GHz. In the probiotic experiment, the cruciform chip could have a 0.72 GHz shift at a concentration of 0.025 mg/50 µL, confirming that terahertz radiation combined with a metamaterial microfluidic chip can perform low-concentration detection.

Terahertz Combined with Metamaterial Microfluidic Chip for Troponin Antigen Detection.

In this paper, we use terahertz combined with metamaterial technology as a powerful tool to identify analytes at different concentrations. Combined with the microfluidic chip, the experimental measurement can be performed with a small amount of analyte. In detecting the troponin antigen, surface modification is carried out by biochemical binding. Through the observation of fluorescent antibodies, the average number of fluorescent dots per unit of cruciform metamaterial is 25.60, and then, by adjusting the binding temperature and soaking time, the average number of fluorescent dots per unit of cruciform metamaterial can be increased to 181.02. Through the observation of fluorescent antibodies, it is confirmed that the antibodies can be successfully stabilized on the metamaterial and then bound to the target antigen. The minimum detectable concentration is between 0.05~0.1 µg/100 µL, and the concentration and ΔY show a positive correlation of R2 = 0.9909.

DNA Sequencing from Subcritical Concentration of Cell-Free DNA Extracted from Electrowetting-on-Dielectric Platform.

Electro-Wetting-On-Dielectric (EWOD) based digital operations have demonstrated outstanding potential in actuating and manipulating liquid droplets. Here, we adapted the EWOD for extracting femtogram quantities of cell-free DNA (cf-DNA) from 1 µL of KSOM mouse embryo culture medium. Our group extracted the femtogram quantity of cf-DNA from 1 µL of mouse embryo culture medium in our previous work. Here, we initially explain a modification from our previous extraction protocol, which improves the extraction percentage to 36.74%. Though the modified extraction protocol improves the extraction percentage from our previously reported work, the quantity is still in the femtogram range. The cf-DNA in femtogram quantity is in subcritical/subthreshold concentration for any further analysis, such as sequencing. To the best of our knowledge, we need a minimum of picogram/nanogram DNA quantities for further analysis. We demonstrated a ground-breaking mechanism of this subcritical concentration of cf-DNA amplification to the nanogram range and performed DNA sequencing. Basic Local Alignment Search Tool (BLAST) is used as a sequence similarity search program to confirm the identity percentage between query and subject. More than 97% of nucleotide identities between query and subject sequences have been obtained from the sequencing result. Hence, we can use the methodology to amplify the subcritical concentration of extracted DNA for further analytics. Moreover, as we extract the cf-DNA from the embryo culture medium, the natural growth of the embryo has not been disrupted. This entire mechanism will pave a new path towards the lab-on-a-chip (LOC) concept.

Virtual Stencil for Patterning and Modeling in a Quantitative Volume Using EWOD and DEP Devices for Microfluidics.

Applying microfluidic patterning, droplets were precisely generated on an electrowetting-on-dielectric (EWOD) chip considering these parameters: number of generating electrodes, number of cutting electrodes, voltage, frequency and gap between upper and lower plates of the electrode array on the EWOD chip. In a subsequent patterning experiment, an environment with three generating electrodes, one cutting electrode and a gap height 10 µm, we obtained a quantitative volume for patterning. Propylene carbonate liquid and a mixed colloid of polyphthalate carbonate (PPC) and photosensitive polymer material were manipulated into varied patterns. With support from a Z-axis lifting platform and a UV lamp, a cured 3D structure was stacked. Using an EWOD system, a multi-layer three-dimensional structure was produced for the patterning. A two-plate EWOD system patterned propylene carbonate in a quantitative volume at 140 Vpp/20 kHz with automatic patterning.

Utilization of a Gas-Sensing System to Discriminate Smell and to Monitor Fermentation during the Manufacture of Oolong Tea Leaves.

The operational duration of shaking tea leaves is a critical factor in the manufacture of oolong tea; this duration influences the formation of its flavor and fragrance. The current method to control the duration of fermentation relies on the olfactory sense of tea masters; they monitor the entire process through their olfactory sense, and their experience decides the duration of shaking and setting. Because of this human factor and olfactory fatigue, it is difficult to define an optimum duration of shaking and setting; an inappropriate duration of shaking and setting deteriorates the quality of the tea. In this study, we used metal-oxide-semiconductor gas sensors to establish an electronic nose (E-nose) system and tested its feasibility. This research was divided into two experiments: distinguishing samples at various stages and an on-line experiment. The samples of tea leaves at various stages exhibited large differences in the level of grassy smell. From the experience of practitioners and from previous research, the samples could be categorized into three groups: before the first shaking (BS1), before the shaking group, and after the shaking group. We input the experimental results into a linear discriminant analysis to decrease the dimensions and to classify the samples into various groups. The results show that the smell can also be categorized into three groups. After distinguishing the samples with large differences, we conducted an on-line experiment in a tea factory and tried to monitor the smell variation during the manufacturing process. The results from the E-nose were similar to those of the sense of practitioners, which means that an E-nose has the possibility to replace the sensory function of practitioners in the future.