Human stem cell-derived cells and tissues hold considerable potential for applications in regenerative medicine, disease modeling and drug discovery. The generation, culture and differentiation of stem cells in low-volume, automated and parallelized microfluidic chips hold great promise to accelerate the research in this domain. Here, we show that we can differentiate human embryonic stem cells (hESCs) to early cardiac mesodermal cells in microfluidic chambers that have a volume of only 30 nanoliters, using discontinuous medium perfusion. 64 of these chambers were parallelized on a chip which contained integrated valves to spatiotemporally isolate the chambers and automate cell culture medium exchanges. To confirm cell pluripotency, we tracked hESC proliferation and immunostained the cells for pluripotency markers SOX2 and OCT3/4. During differentiation, we investigated the effect of different medium perfusion frequencies on cell reorganization and the expression of the early cardiac mesoderm reporter MESP1mCherry by live-cell imaging. Our study demonstrates that microfluidic technology can be used to automatically culture, differentiate and study hESC in very low-volume culture chambers even without continuous medium perfusion. This result is an important step towards further automation and parallelization in stem cell technology.

In order to improve the reliability, safety and whole digestive applicability of the gastrointestinal microrobot (GMR), a novel inchworm-like GMR is proposed in this paper. The expanding mechanism of the robot adopts an overlapping expanding arm structure. This structure increases the variable diameter ratio (ratio of fully expanded diameter to fully folded diameter) of the robot to 3.3, making the robot more applicable to the intestines in various parts of the human body. The mechanical model of the expanding arm is established, and the expanding force at different expanding radii is obtained. And then the expanding force is tested by a force test platform. The force test results: the maximum expanding force is 6.5N, and the minimum expanding force is 1.3N. The trend of the experimental and theoretical values is the same, and the experimental value is less than the theoretical value. A position limiting device based on Hall sensor is designed, which detects whether the mechanism reaches the limit position by non-contact method. This device alleviates the problem of sharp voltage drop caused by motor stall and improves the stability of the control circuit. The results of the Hall-type position limiting device (HPLD) testing show that the working currents of the expanding mechanism and the telescoping mechanism with HPLD are respectively 0.066A and 0.110A, and the robot control circuit works stably. Finally, the robot is tested in the intestine of the living pig, and the safety and reliability of the robot are verified. However, due to the decrease of the efficiency of wireless power transmission in vivo experiments and the change of the position of the receiving coil relative to the transmitting coil, sometimes the power supply is insufficient.

The main objective of this work was to assess the cytotoxic activity of Au/Pt/ZnO nanoparticles synthesized using Arctium lappa extract against leukemia. The Au/Pt/ZnO nanoparticles obtained as a result of biological synthesis were characterized by UV-Vis, Scanning (SEM) and Transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), and Atomic Force Microscopy (AFM). The applied methods showed that the size of nanoparticles ranged from 10 to 40 nm. This work also assessed the cytotoxicity of Au/Pt/ZnO nanoparticles by means of MTT assay, and analyzed apoptosis as well as the influence of the cultivation time and concentration of Au/Pt/ZnO nanoparticles on the percentage of dead cells. The studies showed that the percentage of dead leukemia cells increased with the cultivation time and concentration of Au/Pt/ZnO nanoparticles. There was observed an increase in the percentage of cells in the G2/M phase, which suggests the stoppage of G2/M leading to cell death. The cytotoxicity of Au/Pt/ZnO nanoparticles determined by means of the MTT test indicated that the viability of leukemia cells practically disappeared when the concentration of the tested nanoparticles was 10 mol.

We present a disposable polymeric microfluidic device capable of reversibly binding and purifying Salmonella DNA through solid phase extraction (SPE). The microfluidic channels are first oxygen plasma treated and simultaneously micro-nanotextured, and then functionalized with amine groups via modification with L-histidine or poly-L-histidine. L-Histidine and poly-L-histidine bind on the plasma treated chip surface, and are not detached when rinsing with DNA purification protocol buffers. A pH-dependent protocol is applied on-chip to purify Salmonella DNA, which is first bound on the protonated amines at a pH (5.0) lower than their pKa of surface amine-groups which is 6.0 and then released at a pH higher than the pKa value (10.5). It was found that modification with poly-L-histidine resulted in higher surface density of amine groups onto microfluidic channel. Using the chip modified with poly-L-histidine, high recovery efficiency of at least 550ng of isolated Salmonella DNA as well as DNA purification from Salmonella cell lysates corresponding to less than 5000 cells or 0.026ng of Salmonella DNA was achieved. The protocol developed does not require ethanol or chaotropic solutions typically used in DNA purification, which are known inhibitors for downstream operations such as polymerase chain reactions (PCR) and which can also attack some polymeric microfluidic materials. Therefore, the microfluidic device and the related protocol hold promise for facile incorporation in microfluidics and Lab-on-a-chip (LOC) platforms for pathogen detection or in general for DNA purification.

Deterministic lateral displacement (DLD) is a microfluidic technique for size fractionation of particles/cells in continuous flow with a great potential for biological and clinical applications. Growing interest of DLD devices in enabling high-throughput operation for practical applications, such as circulating tumor cell (CTC) separation, necessitates employing higher flow rates, leading to operation at moderate to high Reynolds number (Re) regimes. Recently, it has been shown that symmetric airfoil shaped pillars with neutral angle-of-attack (AoA) can be used for high-throughput design of DLD devices due to their mitigation of vortex effects and preservation of flow symmetry under high Re conditions. While high-Re operation with symmetric airfoil shaped pillars has been established, the effect of AoAs on the DLD performance has not been investigated. In this paper, we have characterized the airfoil DLD device with various AoAs. The transport behavior of microparticles has been observed and analyzed with various AoAs in realistic high-Re. Furthermore, we have modeled the flow fields and anisotropy in a representative airfoil pillar array, for both positive and negative AoA configurations. Unlike the conventional DLD device, lateral displacement has been suppressed with +5° and + 15° AoA configurations regardless of particle sizes. On the other hand, stronger lateral displacement has been seen with -5° and - 15° AoAs. This can be attributed to growing flow anisotropy as Re climbs, and significant expansion or compression of streamlines between airfoils with AoAs. The findings in this study can be utilized for the design and optimization of airfoil DLD microfluidic devices with various AoAs.

The quantitative studies of cell proliferation and migration under different chemical environments are important for both scientists and clinicians searching for new therapeutics. In this study, we developed a new device to pattern several types of cells in 24-well-plate and demonstrated its' application in cancer cell proliferation and migration assay. The new device combined 3D-printed-silica-part for multi cell types loading with PDMS-through-hole-layer-part for cell micro-patterning which was matched with commercial 24-well-plate. This 24-well-plate based device is flexible and feasible in many applications and can be used in one piece or multi pieces. Besides the application for two types of cells proliferation and migration assay in one chemical condition, as a demonstration, the migration behaviors of four types of cells under 24 types of EGF + bFGF combinatorial conditions were studied. We believed this device could be widely used in clinical searching for new anti-cancer therapeutics and other related studies.

We present a chip design allowing rapid and robust lipid bilayer (LBL) membrane formation using a Parylene coated thin silicon nitride aperture. After bilayer formation, single membrane channels can be reconstituted and characterized by electrophysiology. The ability for robust reconstitution will allow parallelization and enhanced screening of small molecule drugs acting on or permeating across the membrane channel. The aperture was realized on a microfabricated silicon nitride membrane by using standard clean-room fabrication processes. To ensure the lipid bilayer formation, the nitride membrane was coated with a hydrophobic and biocompatible Parylene layer. We tested both Parylene-C and Parylene-AF4. The contact angle measurements on both Parylene types showed very good hydrophobic properties and affinity to lipids. No precoating of the Parylene with an organic solvent is needed to make the aperture lipophilic, in contradiction to Teflon membranes. The chips can be easily placed in an array utilizing a 3D printed platform. Experiments show repetitive LBL formation and destruction (more than 6 times) within a very short time (few seconds). Through measurements we have established that the LBL layers are very thin. This allows the investigation of the fusion process of membrane proteins i.e. outer membrane protein (OmpF) in the LBL within a few minutes.

Glucose and urea enzymatic biosensors were fabricated. One-step electrochemical immobilization process was used to produce thin polyaniline films with entrapped enzymes. Chronopotentiometric analysis, scanning electron microscopy, electrochemical impedance spectroscopy and optical reflectance spectroscopy were used to determine the structure-property relationship of the functionalized polymeric thin films. The device has a recognition stage connected to a potentiometric field-effect-transistor stage and is based on the measurement of microenvironment pH variation or locally produced ions. Optimization of biosensor fabrication and effective measurement conditions were performed. The optimized films presented sensitivity, linearity and detection range to glucose of 14.6 ± 0.4mV/decade, 99.8% and from 10-4M to 10-1mol/L. Two different biosensors were produced based on the enzymatic reaction of urea with selectivity to ammonium or hydroxyl ions. For ammonium ion selective film, the sensor's parameters were 14.7 ± 0.9mV/decade, 98.2% and from 10-5 to 10-1mol/L. For the hydroxyl ion selective film, the same parameters were 7.4 ± 0.5mV/decade, 98.1% and from 10-5 to 10-1mol/L. The change in the oxidation state of the polymeric matrix explains: i) the large loss of functionality of glucose biosensor in time, ii) the conservation of functionality to the hydroxyl ions for urea biosensor and iii) the selectivity variation of the ammonium ion selective urea biosensor. The results indicate that the polymeric matrix has indeed changeable selectivity, what can be applied in different situations for biosensors production.

Overdrainage in upright position is one of the most prevalent issues in treating hydrocephalus with a cerebrospinal fluid (CSF) shunt. Anti-siphon devices (ASDs) are employed to reduce this problem. A novel microelectromechanical system (MEMS)-based valve, termed Chronoflow device, aims to regulate CSF drainage indifferently of the body posture. With this study, the suitability of this MEMS-based valve is evaluated regarding its use for the treatment of hydrocephalus, particularly for the prevention of overdrainage and blockage. In total, four Chronoflow devices were tested. An established in-vitro hardware-in-the-loop (HIL) test bed was used to investigate the valves regarding their pressure-flow characteristics, their behaviors towards CSF dynamics, and their capabilities to prevent CSF overdrainage in upright position. Additionally, a contamination test was conducted to evaluate the susceptibility of the device to blockage due to particles. All valves tested regulated the drainage rate at similar nominal flows and independently of posture. The pressure-flow relation measured, however, was notably higher than numerically calculated. Regarding the CSF dynamics, the first three valves tested led to a decreased steady-state intracranial pressure in supine position and showed stable drainage rate in upright position. During the transitional phase from supine to upright and vice versa, the valves continuously adjusted the outflow resistance, which resulted in a stable transitional phase preventing overdrainage. Yet, the fourth valve showed continuous overdrainage in upright position due to an increased nominal flow. However, after several test iterations the nominal flow decreased and stabilized at a level similar to that of the first three valves tested. The contamination test showed that most particles initially adhere to the pillars and spread throughout the cavity of the valve as the concentration of particles increases, thereby affecting the displacement of the membrane. The devices generally provide a stable flow regulation and prevent overdrainage in upright position. Specifically, their drainage behaviors during the posture changes are very effective. However, they also showed high hysteresis and sensitivity towards particle contamination, which resulted in initial increased and altering nominal flows after many test iterations. This result suggests that the MEMS design presented lacks robustness. Yet, an upstream filter and specific coatings on the fluid pathway may increase significantly its reliability.

In the current study, an electrochemical biosensing signal amplification system was utilized with thionine-chitosan-gold nanoparticles (Chit-GNPs) that absorbed horseradish peroxidase (HRP) and anti-His tagged protein monoclonal antibody derived from Balb/c mice. In addition, transmission electron microscopy (TEM) was used to characterize the nanogold solution and atomic force microscopy (AFM) was used to characterize the sensor assembly. To evaluate the quality of the immunosensor, the amperometric I-t curve method was applied to determine His-IL23 in PBS. The results indicated that the response current exhibited an optimal linear correlation with the His-IL23 concentration that ranged from 0.01 to 103ng/ml. The lowest detection limit was noted at 3.3pg/ml (S/N = 3). The linear equation was deduced as follows: △I = 0.02lgC + 0.037 (R2 = 0.9628). Moreover, it was validated with high sensitivity, reproducibility and rapid response. Apparently, the immunosensor may be a very useful tool for the detection and quantification of His-tagged proteins. In addition, the signal amplification system can be used for the preparation of other immunosensors and to assist in bioassays.