This paper proposes a simple and robust method for spontaneously digitizing aqueous samples into a high-density microwell array. The method is based on an oil-triggered template-confined dewetting phenomenon. To realize the dewetting-induced sample digitization, an aqueous sample is first infused into a networked microwell array (NMA) through a pre-degassing-based self-pumping mechanism, and an immiscible oil phase is then applied over the surface of NMA chip to induce the templated dewetting. Due to periodic interfacial tension heterogeneity, such dewetting ruptures the sample at the thinnest parts (i.e., connection channels) and spontaneously splits the sample into droplets in individual microwells. Without requiring any complex pumping or valving systems, this method can discretize a sample into tens of thousands of addressable droplets in a matter of minutes with nearly 98% usage. To demonstrate the utility and universality of this self-digitization method, we exploited it to discretize samples into 40 233 wells for a digital PCR assay, the digital quantification of bacteria, the self-assembly of spherical colloidal photonic crystals, and the spherical crystallization of drugs. We believe this facile technique will provide a substantial benefit to many compartmentalized assays or syntheses where it is necessary to partition samples into a large number of small individual volumes.
Angiogenesis serves a key role in tumor growth and metastasis. VX-680, a potent inhibitor targeting the Aurora kinase family, is widely used in the inhibition of tumor progression. However, the effect of VX-680 on angiogenesis remains unknown. The present study identified that VX-680 inhibited human umbilical vein endothelial cell (HUVEC) proliferation and promoted HUVEC apoptosis by inducing the cleavage of PARP and caspase-3. VX-680 also markedly decreased the migration and tube formation of HUVECs in a dose-dependent manner. In addition, VX-680 significantly suppressed the formation of blood vessels in a dose-dependent manner confirmed by a chicken embryo chorioallantoic membrane assay in vivo. Furthermore, VX-680 inhibited the expression levels of vascular endothelial growth factor and phosphorylated RAC-α serine/threonine-protein kinase in HUVECs. These results suggested that VX-680 suppressed angiogenesis and may be a potential novel anti-angiogenic agent.
Mycoplasma pneumonia (MP) is a common respiratory infection generally treated with macrolides, but resistance mutations against macrolides are often detected in mycoplasma pneumoniae in China. Rapid and accurate identification of mycoplasma pneumoniae and its mutant type is necessary for precise medication. This paper presents a 3D-printed microfluidic device to achieve this. By 3D printing, the stereoscopic structures such as microvalves, reservoirs, drainage tubes, and connectors were fabricated in one step. The device integrated commercial polymerase chain reaction (PCR) tubes as PCR chambers. The detection was a sample-to-answer procedure. First, the sample, a PCR mix, and mineral oil were respectively added to the reservoirs on the device. Next, the device automatically mixed the sample with the PCR mix and evenly dispensed the mixed solution and mineral oil into the PCR chambers, which were preloaded with the specified primers and probes. Subsequently, quantitative real-time PCR (qPCR) was carried out with the homemade instrument. Within 80 min, mycoplasma pneumoniae and its mutation type in the clinical samples were determined, which was verified by DNA sequencing. The easy-to-make and easy-to-use device provides a rapid and integrated detection approach for pathogens and antibiotic resistance mutations, which is urgently needed on the infection scene and in hospital emergency departments.
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