Single Cell Metabolite Detection Using Inertial Microfluidics-Assisted Ion Mobility Mass Spectrometry.

Single-cell metabolite measurement remains highly challenging due to difficulties related to single cell isolation, metabolite detection, and identification of low levels of metabolites. Here, as a first step of the technological development, we propose a novel strategy integrating spiral inertial microfluidics and ion mobility mass spectrometry (IM-MS) for single-cell metabolite detection and identification. Cells in methanol suspension are inertially focused into a single stream in the spiral microchannel. This stream of separated cells is delivered to the nanoelectrospray needle to be lysed and ionized and subsequently analyzed in real time by IM-MS. This analytical system enables six to eight single-cell metabolic fingerprints to be collected per minute, including gas-phase collisional cross section (CCS) measurements as an additional molecular descriptor, giving increased confidence in metabolite identification. As a proof of concept, the metabolic profiles of three types of cancer cells (U2OS, HepG2, and HepG2.215) were successfully screened, and 19 distinct lipids species were identified with CCS value filtering. Furthermore, principal component analysis (PCA) showed differentiation of the three cancer cell lines, mainly due to cellular surface phospholipids. Taken together, our technology platform offers a simple and efficient method for single-cell lipid profiling, with additional ion mobility separation of lipids significantly improving the confidence toward identification of metabolites.

Paraffin Oil Based Soft-Template Approach to Fabricate Reusable Porous PDMS Sponge for Effective Oil/Water Separation.

Three-dimensional porous material holds enormous potential in the field of life science and environmental protection. In this work, we proposed a facile route for the large-scale synthesis of porous poly(dimethylsiloxane) (PDMS) sponge via paraffin oil based emulsion technique. A stable emulsion could be formed by emulsifying water in the PDMS solution with the aid of paraffin oil. Moreover, the amount of emulsified water in 5 g of PDMS solution is as high as 35 g, and the skeleton of the prepared PDMS sponge is still in intact. This method is cost-effective, rapid, and easily scaled up. The water contact angle of the obtained PDMS sponge is 141.9 ± 1°, and the absorption capacities of the sponges are 13.5-33.3 g g-1 for various organic solvents. The PDMS sponge only needed 0.069 MPa force to realize the compression ratio of 90%, which it still maintained after over 50 cycles of compression. In addition, the porous PDMS sponge exhibited an excellent oil absorption and an outstanding reusability, which were potentially useful in water purification.

A High-Capacity Polyethylene Oxide-Based All-Solid-State Battery Using a Metal-Organic Framework Hosted Silicon Anode.

Polyethylene oxide (PEO)-based solid electrolytes have been widely studied in all-solid-state lithium (Li) metal batteries due to their favorable interfacial contact with electrodes, facile fabrication, and low cost, but their inferior Li dendrite suppression capability renders low actual areal capacities of Li metal anodes. Here, we develop a high-capacity all-solid-state battery using a metal-organic framework hosted silicon (Si@MOF) anode and a fiber-supported PEO/garnet composite electrolyte. Si nanoparticles are embedded in the micro-sized MOF-derived carbon host, which efficiently accommodates the repeated deformation of Si over cycles while providing sufficient charge transfer pathways. As a result, the Si@MOF anode shows excellent interfacial stability toward the composite polymer electrolyte for over 1000 h and achieves a high reversible areal capacity of 3 mAh cm-2. The full cell using the LiFePO4 (LFP) cathode is able to deliver 135 mAh g-1 initially and maintains 73.1% of the capacity after 500 cycles at 0.5 C and 60 °C. More remarkably, the full cells with high LFP loadings achieve areal capacities of more than 2 mAh cm-2, exceeding most PEO-based ASSBs using metallic Li. Finally, the pouch cell using the proposed design exhibits decent electrochemical performance and high safety.

A high-energy and long-cycling lithium-sulfur pouch cell via a macroporous catalytic cathode with double-end binding sites.

Lithium-sulfur batteries are attractive alternatives to lithium-ion batteries because of their high theoretical specific energy and natural abundance of sulfur. However, the practical specific energy and cycle life of Li-S pouch cells are significantly limited by the use of thin sulfur electrodes, flooded electrolytes and Li metal degradation. Here we propose a cathode design concept to achieve good Li-S pouch cell performances. The cathode is composed of uniformly embedded ZnS nanoparticles and Co-N-C single-atom catalyst to form double-end binding sites inside a highly oriented macroporous host, which can effectively immobilize and catalytically convert polysulfide intermediates during cycling, thus eliminating the shuttle effect and lithium metal corrosion. The ordered macropores enhance ionic transport under high sulfur loading by forming sufficient triple-phase boundaries between catalyst, conductive support and electrolyte. This design prevents the formation of inactive sulfur (dead sulfur). Our cathode structure shows improved performances in a pouch cell configuration under high sulfur loading and lean electrolyte operation. A 1-A-h-level pouch cell with only 100% lithium excess can deliver a cell specific energy of >300 W h kg-1 with a Coulombic efficiency >95% for 80 cycles.

Agarose-based microwell array chip for high-throughput screening of functional microorganisms.

Natural bacterial communities provide a rich source of biocatalysts, however, high-throughput screening for the functional bacteria remains a major challenge. Here, we present an agarose-based microwells array chip for functionally screening and isolating novel microorganisms with merits of high-throughput, high specificity and sensitivity. In this approach, the bacterial cells were loaded with single cell per a microwell mode and were incubated in the arrayed agarose microwells. The growths of single cells were then monitored in real time and the enzyme reaction activities were assessed at the individual microwell resolution. To validate the reliability of the method, we subsequently applied it to screen lipase-producing bacteria from the pond water based on lipase hydrolysis of the substrate in the presence of a fluorescent dye (rhodamine B), which emitted yellow-orange fluorescent halos upon UV-light irradiation. A total amount of more than 310,000 isolates from pond water could be screened at a time in only 13 h with reduced consumption of reagents. As a result, 12 microcolonies were identified out with the desired phenotype, in which two new species were discovered based on 16S rRNA sequencing. We expect the developed method to be potentially useful to high-throughput analysis, microbiology and synthetic biology.

Droplet Jumping: Effects of Droplet Size, Surface Structure, Pinning, and Liquid Properties.

Coalescence-induced droplet jumping has the potential to enhance the efficiency of a plethora of applications. Although binary droplet jumping is quantitatively understood from energy and hydrodynamic perspectives, multiple aspects that affect jumping behavior, including droplet size mismatch, droplet-surface interaction, and condensate thermophysical properties, remain poorly understood. Here, we develop a visualization technique utilizing microdroplet dispensing to study droplet jumping dynamics on nanostructured superhydrophobic, hierarchical superhydrophobic, and hierarchical biphilic surfaces. We show that on the nanostructured superhydrophobic surface the jumping velocity follows inertial-capillary scaling with a dimensionless velocity of 0.26 and a jumping direction perpendicular to the substrate. A droplet mismatch phase diagram was developed showing that jumping is possible for droplet size mismatch up to 70%. On the hierarchical superhydrophobic surface, jumping behavior was dependent on the ratio between the droplet radius Ri and surface structure length scale L. For small droplets ( Ri ≤ 5 L), the jumping velocity was highly scattered, with a deviation of the jumping direction from the substrate normal as high as 80°. Surface structure length scale effects were shown to vanish for large droplets ( Ri > 5 L). On the hierarchical biphilic surface, similar but more significant scattering of the jumping velocity and direction was observed. Droplet-size-dependent surface adhesion and pinning-mediated droplet rotation were responsible for the reduced jumping velocity and scattered jumping direction. Furthermore, droplet jumping studies of liquids with surface tensions as low as 38 mN/m were performed, further confirming the validity of inertial-capillary scaling for varying condensate fluids. Our work not only demonstrates a powerful platform to study droplet-droplet and droplet-surface interactions but provides insights into the role of fluid-substrate coupling as well as condensate properties during droplet jumping.

Chemically Edited Exosomes with Dual Ligand Purified by Microfluidic Device for Active Targeted Drug Delivery to Tumor Cells.

Exosomes, which are lipid membrane-bound nanovesicles (50-150 nm in diameter), have aroused extensive attention for their potential applications in invasive molecular and stand for a new therapeutic delivery system. However, they are limited by poor targeting ability and a lack of efficient isolation techniques. Here, we present a three-dimensional nanostructured microfluidic chip, in which arrays of micropillars were functionalized with crisscrossed multiwall carbon nanotubes by chemical deposition, to capture exosomes with high efficiency through a combination of a specific recognition molecule (CD63) and the unique topography of the nanomaterials. As is proven, this nanostructured interface substantially made the immuno capturing of exosomes more efficient. A high percentage of intact vesicles <150 nm were readily purified. As a further application, we added functionality to the exosomes by a chemical editing approach for targeted drug delivery. Donor cells were labeled chemically with dual ligands (biotin and avidin) in the phospholipid membrane and encapsulated drugs in the cytosol. Though the engineered donor cells secreted exosomes, the dual ligands, together with the drugs, were inherited by the exosomes, which were then isolated with the microfluidic chip. Then, the isolated exosomes were used as drug delivery vehicles and showed strong targeting abilities to tumor cells and highly efficient receptor-mediated cellular uptake when exposed to recipient cells. Thus, the anticancer effect of chemotherapeutic drugs was improved significantly. It suggested that this platform could provide a useful tool for isolating intact exosomes with high efficiency and exploiting their natural carrier function to deliver chemotherapeutic drugs to tumor cells with increased efficacy and targeting capacity.

The use of RGD-engineered exosomes for enhanced targeting ability and synergistic therapy toward angiogenesis.

Promoted therapeutic angiogenesis is a major objective in the area of regenerative medicine, and sufficient vascularization of artificial tissues or organs is one of the main difficulties for the realization of tissue engineering methods. The identification of new kinds of pro-angiogenic materials will greatly profit developments in regenerative medicine. The use of exosomes for this intention is a considerably new idea developed in recent years. However, several limitations need to be addressed before their use as clinical therapeutics, including the lack of efficient exosome enrichment and methods to endow exosomes with targeting ability. Herein, we pioneered biomimetic particles with topographic structures for exosome isolation. Using this system, nearly 80% of exosomes were isolated in 30 min. Through a donor cell-assisted membrane modification strategy, the isolated exosomes exhibited increased targeting to blood vessels due to the modified Arg-Gly-Asp (RGD) peptide on the exosome membrane, and simultaneously possessed a synergistic therapeutic angiogenesis effect and angiogenesis imaging attributed to metabolic labeling by click chemistry both in vitro and in vivo. The engineered exosomes represent a potential new therapeutic tool for angiogenesis therapy and imaging in a bio-friendly manner.