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The selective cell separation is a critical step in fundamental life sciences, translational medicine, biotechnology, and energy harvesting. Conventional cell separation methods are fluorescent activated cell sorting and magnetic-activated cell sorting based on fluorescent probes and magnetic particles on cell surfaces. Label-free cell separation methods such as Raman-activated cell sorting, electro-physiologically activated cell sorting, dielectric-activated cell sorting, or inertial microfluidic cell sorting are, however, limited when separating cells of the same kind or cells with similar sizes and dielectric properties, as well as similar electrophysiological phenotypes. Here we report a label-free density difference amplification-based cell sorting (dDACS) without using any external optical, magnetic, electrical forces, or fluidic activations. The conceptual microfluidic design consists of an inlet, hydraulic jump cavity, and multiple outlets. Incoming particles experience gravity, buoyancy, and drag forces in the separation chamber. The height and distance that each particle can reach in the chamber are different and depend on its density, thus allowing for the separation of particles into multiple outlets. The separation behavior of the particles, based on the ratio of the channel heights of the inlet and chamber and Reynolds number has been systematically studied. Numerical simulation reveals that the difference between the heights of only lighter particles with densities close to that of water increases with increasing the ratio of the channel heights, while decreasing Reynolds number can amplify the difference in the heights between the particles considered irrespective of their densities. [ABSTRACT FROM AUTHOR]

Just as the Petri dish has been invaluable to the evolution of biomedical science in the last 100 years, microfluidic cell assay platforms have the potential to change significantly the way modern biology and clinical science are performed. However, an evolutionary process of creating an efficient microfluidic array for many different bioassays is necessary. Specifically for a complete view of a cell response it is essential to incorporate cytotoxic, protein and gene analysis on a single system. Here we present a novel cellular and molecular analysis platform, which allows access to gene expression, protein immunoassay, and cytotoxicity information in parallel. It is realized by an integrated microfluidic array plate (iMAP). The iMAP enables sample processing of cells, perfusion based cell culture, effective perturbation of biologic molecules or drugs, and simultaneous, real-time optical analysis for different bioassays. The key features of the iMAP design are the interface of on-board gravity driven flow, the open access input fluid exchange and the highly efficient sedimentation based cell capture mechanism (∼100% capture rates). The operation of the device is straightforward (tube and pump free) and capable of handling dilute samples (5-cells per experiment), low reagent volumes (50 nL per reaction), and performing single cell protein and gene expression measurements. We believe that the unique low cell number and triple analysis capabilities of the iMAP platform can enable novel dynamic studies of scarce cells. [ABSTRACT FROM AUTHOR]

Highly selective and sensitive optical methods for the detection of metal ions have had a substantial impact on molecular biology, environmental monitoring and other areas of research. Here we demonstrate a new method for detecting metal ions that is based on selective plasmonic resonance energy transfer (PRET) between conjugated metal–ligand complexes and a single gold nanoplasmonic probe. In addition to offering high spatial resolution due to the small size of the probe, our method is 100 to 1,000 times more sensitive than organic reporter-based methods. Moreover, it can achieve high selectivity owing to the selective formation of Cu2+ complexes and selective resonant quenching of the gold nanoplasmonic probe by the conjugated complexes. We expect that PRET-based metal ion sensing could have applications in cellular imaging, systems biology and environmental monitoring. [ABSTRACT FROM AUTHOR]

In this paper, signal amplifications of surface-enhanced Raman scattering (SERS) are realized by an optofluidic compact disc (CD)-based preconcentration method for effective label-free environmental and biomolecular detections. The preconcentration of target molecules is accomplished through the accumulation of adsorbed molecules on SERS-active sites by repeating a ‘filling–drying’ cycle of the assay solution in the optofluidic CD platform. After 30 cycles, the clear and high SERS signal of rhodamine 6G of 1 nM is readily detected. In addition to the preconcentration-based signal amplification by the optofluidic SERS system on the CD platform, we introduce a controlled precipitation of gold nanoparticles by CuSO4for SERS substrates. This method provides high-throughput, high-sensitive and large-area uniform SERS substrates on the optofluidic CD platform. The uniform SERS signals from different positions in spots of 3 mm2on the different CDs gives us confidence in the reliability and stability of our SERS substrates. [ABSTRACT FROM AUTHOR]

We describe the development of innovative plasmon resonance energy transfer (PRET)-based molecular imaging of biomolecules in living cells. Our strategy of invivo PRET imaging relies on the resonant plasmonic energy transfer from a gold nanoplasmonic probe to conjugated target molecules, which creates “quantized quenching dips” within the Rayleigh scattering spectrum of the probe. The positions of these quantized quenching dips exactly match with the absorption peaks of the target molecule since we intentionally design nanoantennas (i.e., nanoplasmonic probes) to overlap the electronic dipoles of the molecule and the plasmonic resonance dipole of nanoantennas. Such the quenching dips allow quantitative and long-term dynamic imaging of the target molecule without the drawbacks of photobleaching and blinking inherent to fluorescent markers, which cannot provide chemical fingerprints. Compared with other imaging methods, our PRET spectroscopic imaging method allows us to generate nanoscale specific wavelengths of local light sources in living systems via nanoantennas and transmit back the nanospectroscopic imaging data of biochemical activities in living cells. As a first demonstration of invivo PRET imaging, we performed a visualization of the dynamics of intracellular cytochrome cin HepG2 cells under ethanol-induced apoptosis. [ABSTRACT FROM AUTHOR]

Mesoporous materials have attracted considerable attention for use as a catalyst or a catalyst support due to their remarkable textural properties such as high surface area and large pore volume with a narrow pore size distribution. Many efforts have been made to design mesoporous materials for use in heterogeneous catalyst systems. Recent progress and results regarding the preparation of nickel-mesoporous materials and their application to the hydrodechlorination of chlorinated organic compounds were discussed in this review. Mesoporous materials were used as a support for nickel catalyst or a nickel-incorporated mesoporous catalytic material in this work. Two research areas were described and discussed in this review. One is the preparation of mesoporous alumina-supported nickel catalysts and their application to the hydrodechlorination of 1,2-dichloropropane and o-dichlorobenzene. The other is the preparation of mesoporous silica-supported nickel catalysts and their application to the hydrodechlorination of 1,1,2-trichloroethane and chlorobenzene. [ABSTRACT FROM AUTHOR]

Mitochondrial damage represents a dramatic change in cellular homeostasis. One rapid response is perimitochondrial actin polymerization, termed acute damage-induced actin (ADA). The consequences of ADA are not understood. In this study, we show evidence suggesting that ADA is linked to rapid glycolytic activation upon mitochondrial damage in multiple cells, including mouse embryonic fibroblasts and effector CD8+ T lymphocytes. ADA-inducing treatments include CCCP, antimycin, rotenone, oligomycin, and hypoxia. The Arp2/3 complex inhibitor CK666 or the mitochondrial sodium-calcium exchanger (NCLX) inhibitor CGP37157 inhibits both ADA and the glycolytic increase within 5 min, supporting ADA's role in glycolytic stimulation. Two situations causing chronic reductions in mitochondrial ATP production, mitochondrial DNA depletion and mutation to the NDUFS4 subunit of complex 1 of the electron transport chain, cause persistent perimitochondrial actin filaments similar to ADA. CK666 treatment causes rapid mitochondrial actin loss and a drop in ATP in NDUFS4 knock-out cells. We propose that ADA is necessary for rapid glycolytic activation upon mitochondrial impairment, to re-establish ATP production. [ABSTRACT FROM AUTHOR]

A highly specific interaction between a metal-deficient metalloenzyme and metal ion has been utilized in the selective detection of the metal ion by surface plasmon resonance spectroscopy (SPRS). The use of SPRS and Cu-demetallated superoxide dismutase (E,Zn-SOD1) as a sensing actuator allows one to selectively and in situ detect Cu2+ without any interference that other spectroscopic methods may have. [ABSTRACT FROM AUTHOR]

Interactions between protein aggregates and a cellular membrane have been strongly implicated in many protein conformational diseases. However, such interactions for the case of Cu/Zn superoxide dismutase (SOD1) protein, which is related to fatal neurodegenerative disorder amyotrophic lateral sclerosis (ALS), have not been explored yet. For the first time, we report the direct observation of defect formation and increased ion permeability of a membrane induced by SOD1 aggregates using a supported lipid bilayer and membrane patches of human embryonic kidney cells as model membranes. We observed that aggregated SOD1 significantly induced the formation of defects within lipid membranes and caused the perturbation of membrane permeability, based on surface plasmon resonance spectroscopy, atomic force microscopy and electrophysiology. In the case of apo SOD1 with an unfolded structure, we found that it bound to the lipid membrane surface and slightly perturbed membrane permeability, compared to other folded proteins (holo SOD1 and bovine serum albumin). The changes in membrane integrity and permeability were found to be strongly dependent on the type of proteins and the amount of aggregates present. We expect that the findings presented herein will advance our understanding of the pathway by which structurally disordered SOD1 aggregates exert toxicity in vivo. [ABSTRACT FROM AUTHOR]