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High performance sorting of circulating tumor cells (CTCs) from peripheral blood is key to liquid biopsies. Size-based deterministic lateral displacement (DLD) technique is widely used in cell sorting. But conventional microcolumns have poor fluid regulation ability, which limits the sorting performance of DLD. When the size difference between CTCs and leukocytes is small (e.g., less than 3 μm), not only DLD, many size-based separation techniques fail due to low specificity. CTCs have been confirmed to be softer than leukocytes, which could serve as a basis for sorting. In this study, we presented a multistage microfluidic CTCs sorting method, first sorting CTCs using a size-based two-array DLD chip, then purifying CTCs mixed by leukocytes using a stiffness-based cone channel chip, and finally identifying cell types using Raman techniques. The entire CTCs sorting and analysis process was label free, highly pure, high-throughput and efficient. The two-array DLD chip employed a droplet-shaped microcolumn (DMC) developed by optimization design rather than empirical design. Attributed to the excellent fluid regulation capability of DMC, the CTCs sorter system developed by parallelizing four DMC two-array DLD chips was able to process a sample of 2.5 mL per minute with a recovery efficiency of 96.30 ± 2.10% and a purity of 98.25 ± 2.48%. To isolate CTCs mixed dimensionally by leukocytes, a cone channel sorting method and chip were developed based on solid and hydrodynamic coupled analysis. The cone channel chip allowed CTCs to pass through the channel and entrap leukocytes, improving the purity of CTCs mixed by leukocytes by 1.8-fold. • The multistage microfluidic cell sorting method is the only reported whole microfluidic label free sorting analysis method. • The droplet-shaped microcolumn improved the sorting performance of the deterministic lateral displacement technique. • Developed a CTCs sorter system, and the performance of this system surpasses most microfluidic label free technologies. • Developed a cone channel sorting method and chip, improving the purity of CTCs mixed by leukocytes 1.8-fold. [ABSTRACT FROM AUTHOR]

Graphene is widely recognized as a promising nanomaterial for the construction of high-performance electrochemical biosensors. However, the lack of strong interfacial forces between graphene and conductive substrates is a bottleneck in the fabrication of highly stable graphene electrodes. In this work, few-layer graphene was directly formed on a high pressure high temperature (HPHT) diamond substrate via sp 3 -to-sp 2 conversion by catalytic thermal treatment and using diamond itself as the carbon source. The hybrid electrode prototype was also highly conductive and had a linear electrochemical response to dopamine in the concentration range of 5 μM – 2 mM, with a low detection limit of 200 nM. After prolonged and repeated exposure to dopamine, electrode fouling was observed which led to sensitivity degradation. Based on the strong interfacial bonding between graphene and HPHT diamond, regeneration of the fouled electrode and full performance recovery would be easily achieved by ultrasonic cleaning. The hybrid electrode is highly robust, and shows potential in its application to the detection of biofouling molecules, food processing and wastewater treatment. [ABSTRACT FROM AUTHOR]

Copper (II) is one of the most of important cofactors for numerous enzymes and has captured broad attention due to its role as a neurotransmitters for physiological and pathological functions. In this article, we present a reaction-based fluorescent sensor for Cu 2+ detection ( NIR-Cu ) with near-infrared excitation and emission, including probe design, structure characterization, optical property test and biological imaging application. NIR-Cu is equipped with a functional group, 2-picolinic ester, which hydrolyzes in the presence of Cu 2+ with high selectivity over completed cations. With the experimental conditions optimized, NIR-Cu ( 5 μM ) exhibits linear response for Cu 2+ range from 0.1 to 5 μM, with a detection limit of 29 nM. NIR-Cu also shows excellent water solubility and are highly responsive, both desirable properties for Cu 2+ detection in water samples. In addition, due to its near-infrared excitation and emission properties, NIR-Cu demonstrates outstanding fluorescent imaging in living cells and tissues. [ABSTRACT FROM AUTHOR]

Hydrogen sulfide (H 2 S), one of the biologically important gaseous signal molecules, plays an essential role in diverse normal biochemical functions and pathological processes. Herein, an efficient two-photon in and near-infrared out mitochondria-targeting dye has been designed, synthesized and characterized. It is easily synthesized by the condensation reaction (C˭C) of 4-hydroxybenzaldehyde and 6-(diethylamino)−1,2,3,4-tetrahydroxanthylium (mitochondria-targeting), which possesses large two-photon action absorption cross-section ~160 g and high fluorescence quantum yield ~0.15. Encouraged by the results, we proceeded to conjugate this new dye with a H 2 S recognition moiety (4-dinitrobenzene-ether, DNB), on the basis of the intramolecular charge transfer (ICT) strategy, to construct a novel H 2 S fluorescent probe (TP-NIR-HS), which shows a targeting ability with high sensitivity and selectivity, and low cytotoxicity. This new probe was then applied for two-photon imaging of living cells and tissues and showed high imaging resolution and a deep-tissue imaging depth of ~350 µm, thus demonstrating its practical application in biological systems, and providing a valuable theoretical basis and technical support for the study of physiological and pathological functions of H 2 S. [ABSTRACT FROM AUTHOR]

Abstract: In this study, a microfluidic platform was developed to generate single layer, linear array of microbeads for multiplexed high-throughput analysis of biomolecules. The microfluidic device is comprised of eight microbead-trapping units, where microbeads were immobilized in a linear array format by the exertion of a negative pressure in the control channel connected to each sieving microstructure. Multiplexed assays were achieved by using a mixture of different spectrally-encoded microbeads functionalized with specific probes, followed by on-chip reaction and detection. The microfluidic-based microbeads array platform was employed for multiplexed analysis of DNA and proteins, as demonstrated by the simultaneous discrimination of four HPV genotypes and the parallel detection of six different proteins. Compared with the off-chip protocols, the on-chip analysis exhibited better reaction efficiency, higher sensitivity and wider linear detection range. Visual inspection and identification of functionalized microbeads were facilitated by the single layer arrangement of microbeads so that accurate data acquisition can be performed during the detection process. [Copyright &y& Elsevier]