Gasdermin A Is Required for Epidermal Cornification during Skin Barrier Regeneration and in an Atopic Dermatitis-Like Model.

Atopic dermatitis is featured with impaired skin barrier. The stratum corneum and the intercellular tight junctions constitute the permeability barrier, which is essential to protect water loss in the host and prevent pathogen entry. The epidermal barrier is constantly renewed by differentiating keratinocytes through cornification, during which autophagy contributes to elimination of organelles and nucleus. The human GSDMA and its mouse homologs Gsdma1-3 are expressed in the suprabasal epidermis. Although a pyroptotic role of GSDMA/Gsdma1 in host defense against Streptococcus pyogenes has been reported, the physiological function of Gsdma1/a2/a3 in epidermal homeostasis remains elusive. Here, through repeated epidermal barrier disruption, we found that tight junction formation and stratum corneum maturation were defective in the Gsdma1/a3-deficient epidermis. Using comparative gene profiling analysis, mitochondrial respiration measurement, and in vivo tracing of mitophagy, our data indicate that Gsdma1/a3 activation leads to mitochondrial dysfunction and subsequently facilitates mitochondrial turnover and epidermal cornification. In calcipotriol (MC903)-induced atopic dermatitis-like animal model, we showed that Gsdma1/a3-deficiency selectively enhanced the T helper type 2 response. Remarkably, the GSDMA expression is reduced in the epidermis of patients with atopic dermatitis compared with that of normal individuals. Gsdma1/a3-deficiency might be involved in atopic dermatitis pathogenesis, likely through GSDMA-mediated epidermal differentiation and cornification.

A Low-Power CMOS Microfluidic Pump Based on Travelling-Wave Electroosmosis for Diluted Serum Pumping.

Microfluidic pump is an essential component in lab-on-chip applications. It is of importance to develop an active microfluidic pump with low-power and low-cost characteristics for portable and miniaturized diagnostic systems. Taking advantages of CMOS technologies, in this work, we report a low-power microfluidic pump based on travelling-wave electroosmosis (TWEO). Utilizing an integrated driving circuit, this monolithic CMOS microfluidic pump can be operated at 1.5 V driving voltage with a power consumption of 1.74 mW. The integrated driving circuit consist of a resistor-capacitor (RC) oscillator, a 90-degrees phase-shift square wave generator, and buffer amplifiers. Moreover, capabilities of the developed CMOS TWEO pump to drive diluted human serum are characterized. The flow rate of diluted human serum with dilution ratio of 1:1000 can achieve 51 µm/s. This is the first time demonstrating an in-situ CMOS-based microfluidic pump to drive the clinical diluted serum sample. As a consequence, this work demonstrates an essential component of CMOS biotechnologies for potential applications of portable in vitro diagnosis (IVD) systems.

Local Chemical Stimulation of Neurons with the Fluidic Force Microscope (FluidFM).

Physiological communication between neurons is dependent on the exchange of neurotransmitters at the synapses. Although this chemical signal transmission targets specific receptors and allows for subtle adaptation of the action potential, in vitro neuroscience typically relies on electrical currents and potentials to stimulate neurons. The electric stimulus is unspecific and the confinement of the stimuli within the media is technically difficult to control and introduces large artifacts in electric recordings of the activity. Here, we present a local chemical stimulation platform that resembles in vivo physiological conditions and can be used to target specific receptors of synapses. Neurotransmitters were dispensed using the force-controlled fluidic force microscope (FluidFM) nanopipette, which provides exact positioning and precise liquid delivery. We show that controlled release of the excitatory neurotransmitter glutamate induces spiking activity in primary rat hippocampal neurons, as measured by concurrent electrical and optical recordings using a microelectrode array and a calcium-sensitive dye, respectively. Furthermore, we characterized the glutamate dose response of neurons by applying stimulation pulses of glutamate with concentrations from 0 to 0.5 mm. This new stimulation approach, which combines FluidFM for gentle and precise positioning with a microelectrode array read-out, makes it possible to modulate the activity of individual neurons chemically and simultaneously record their induced activity across the entire neuronal network. The presented platform not only offers a more physiological alternative compared with electrical stimulation, but also provides the possibility to study the effects of the local application of neuromodulators and other drugs.

A low sample volume particle separation device with electrokinetic pumping based on circular travelling-wave electroosmosis.

Particle separation is a crucial step in sample preparation processes. The preparation of low volume samples is especially important for clinical diagnosis and chemical analysis. The advantages of microfluidic techniques have lead them to become potential candidates for particle separation. However, existing microfluidic devices require external pumping sources and extensive geometric patterns to attain high separation efficiency, which is disadvantageous when handling low volume samples. This paper presents a low sample volume particle separation microfluidic device with low voltage electrokinetic pumping based on circular travelling-wave electroosmosis (TWEO). Computational numerical software was utilized to simulate two electrokinetic mechanisms: circular TWEO and dielectrophoresis (DEP). The circular TWEO shear flow generates a velocity gradient in the radial direction which causes a shear stress-induced force to drag particles into the center region of the device. In contrast, the non-parallel electrodes induce negative DEP forces which push polystyrene beads towards the peripheral regions; the magnitude of the DEP forces are dependent on the sizes of the polystyrene beads. We used particles of various sizes to experimentally prove the concept of particle separation. Our experiments show that 15 µm beads are dragged into the center region due to the shear stress-induced force, and 1 µm beads move towards the outer region because of the large negative DEP force. The results show a separation purity of 94.4% and 80.0% for 15 µm and 1 µm beads respectively. We further demonstrated particle isolation from a sample of containing a small proportion of 6 µm beads mixed with 1 µm beads at a concentration ratio of 1 : 300. Therefore, the innovative device developed in this paper provides a promising solution to allow particle separation in sample volumes as low as 50 nL.

A microfluidic device for uniform-sized cell spheroids formation, culture, harvesting and flow cytometry analysis.

Culture of cells as three-dimensional (3D) aggregates, named spheroids, possesses great potential to improve in vitro cell models for basic biomedical research. However, such cell spheroid models are often complicated, cumbersome, and expensive compared to conventional Petri-dish cell cultures. In this work, we developed a simple microfluidic device for cell spheroid formation, culture, and harvesting. Using this device, cells could form uniformly sized spheroids due to strong cell-cell interactions and the spatial confinement of microfluidic culture chambers. We demonstrated cell spheroid formation and culture in the designed devices using embryonic stem cells, carcinoma cells, and fibroblasts. We further scaled up the device capable of simultaneously forming and culturing 5000 spheroids in a single chip. Finally, we demonstrated harvesting of the cultured spheroids from the device with a simple setup. The harvested spheroids possess great integrity, and the cells can be exploited for further flow cytometry assays due to the ample cell numbers.

Single channel layer, single sheath-flow inlet microfluidic flow cytometer with three-dimensional hydrodynamic focusing.

Flow cytometry is a technique capable of optically characterizing biological particles in a high-throughput manner. In flow cytometry, three dimensional (3D) hydrodynamic focusing is critical for accurate and consistent measurements. Due to the advantages of microfluidic techniques, a number of microfluidic flow cytometers with 3D hydrodynamic focusing have been developed in recent decades. However, the existing devices consist of multiple layers of microfluidic channels and tedious fluidic interconnections. As a result, these devices often require complicated fabrication and professional operation. Consequently, the development of a robust and reliable microfluidic flow cytometer for practical biological applications is desired. This paper develops a microfluidic device with a single channel layer and single sheath-flow inlet capable of achieving 3D hydrodynamic focusing for flow cytometry. The sheath-flow stream is introduced perpendicular to the microfluidic channel to encircle the sample flow. In this paper, the flow fields are simulated using a computational fluidic dynamic (CFD) software, and the results show that the 3D hydrodynamic focusing can be successfully formed in the designed microfluidic device under proper flow conditions. The developed device is further characterized experimentally. First, confocal microscopy is exploited to investigate the flow fields. The resultant Z-stack confocal images show the cross-sectional view of 3D hydrodynamic with flow conditions that agree with the simulated ones. Furthermore, the flow cytometric detections of fluorescence beads are performed using the developed device with various flow rate combinations. The measurement results demonstrate that the device can achieve great detection performances, which are comparable to the conventional flow cytometer. In addition, the enumeration of fluorescence-labelled cells is also performed to show its practicality for biological applications. Consequently, the microfluidic flow cytometer developed in this paper provides a practical platform that can be used for routine analysis in biological laboratories. Additionally, the 3D hydrodynamic focusing channel design can also be applied to various applications that can advance the lab on a chip research.

Room temperature current injection polariton light emitting diode with a hybrid microcavity.

The strong light-matter interaction within a semiconductor high-Q microcavity has been used to produce half-matter/half-light quasiparticles, exciton-polaritons. The exciton-polaritons have very small effective mass and controllable energy-momentum dispersion relation. These unique properties of polaritons provide the possibility to investigate the fundamental physics including solid-state cavity quantum electrodynamics, and dynamical Bose-Einstein condensates (BECs). Thus far the polariton BEC has been demonstrated using optical excitation. However, from a practical viewpoint, the current injection polariton devices operating at room temperature would be most desirable. Here we report the first realization of a current injection microcavity GaN exciton-polariton light emitting diode (LED) operating under room temperature. The exciton-polariton emission from the LED at photon energy 3.02 eV under strong coupling condition is confirmed through temperature-dependent and angle-resolved electroluminescence spectra.

Characteristics of exciton-polaritons in ZnO-based hybrid microcavities.

Wide bandgap semiconductors are promising materials for the development of polariton-based optoelectronic devices operating at room temperature (RT). We report the characteristics of ZnO-based microcavities (MCs) in the strong coupling regime at RT with a vacuum Rabi splitting of 72 meV. The impact of scattering states of excitons on polariton dispersion is investigated. Only the lower polariton branches (LPBs) can be clearly observed in ZnO MCs since the large vacuum Rabi splitting pushes the upper polariton branches (UPBs) into the scattering absorption states in the ZnO bulk active region. In addition, we systematically investigate the polariton relaxation bottleneck in bulk ZnO-based MCs. Angle-resolved photoluminescence measurements are performed from 100 to 300 K for different cavity-exciton detunings. A clear polariton relaxation bottleneck is observed at low temperature and large negative cavity detuning conditions. The bottleneck is suppressed with increasing temperature and decreasing detuning, due to more efficient phonon-assisted relaxation and a longer radiative lifetime of the polaritons.