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  • Cox Rossen posted an update 2 days, 7 hours ago

    Circulating tumor cell (CTC) clusters, which are multicellular groups of CTCs, were recently suggested to had the greater potential of forming distal metastasis than single CTCs. However, our understanding of the forming of CTC clusters is still limited since there are few existing methods to study cancer cells aggregation kinetics, especially for a small number of cells. Herein we report a high-throughput miniaturized microwell-based cell aggregation-chip (AG-chip) to enable better characterize of the tumor cells clustering process. We successfully demonstrated the capability of the AG-chip in determining cell aggregation, and found that (1) high metastatic breast cancer cells (MDA-MB-231 & MDA-MB-436) have stronger aggregation capacities than those low metastatic breast cancer cells (MCF-7 & SK-BR-3); (2) cells with similar aggregation ability were distinguished through the analysis of aggregation kinetics; (3) the detected aggregation ability can be used to indicate the metastatic potential of the cells; (4) the inhibition of integrins could regulate the cell clustering via blockage of cell adhesion or/and cell migration. This newly developed microdevice may promote further study of CTC clusters and metastasis.Herein, we report a colorimetric sensing system for the detection of highly virulent bacteria, Escherichiacoli O157H7, in sausage by utilizing magnetic separation and enzyme-mediated signal amplification on paper disc. For magnetic separation, Poly-l-lysine coated starch magnetic particles (PLL@SMPs) were synthesized and utilized for the separation and concentration of the bacteria in sample suspension. Horseradish peroxidase-conjugated antibody (HRP-Antibody) and 3,3′,5,5′- tetramethylbenzidine (TMB) were employed for the specific signal amplification in the presence of target bacteria. The synthesized PLL@SMPs showed an excellent capture efficiency (>90%) for the pathogenic bacteria in large volume sample suspension. The intrinsic problems associated with the non-specific binding of sensing components that lead to the high background signal and low sensitivity in colorimetric detection was successfully resolved by employing hyaluronic acid as a blocking agent. The effective separation and concentration of target bacteria by PLL@SMPs and target-specific signal amplification with exceptionally high signal to noise ratio enabled the detection of target bacteria with a detection limit in the single digit regime. The sensing system proposed in this study was successfully used for the detection of the target pathogenic bacteria, E. coli O157H7, in sausage sample with the limit of detection (LOD) as low as 30.8 CFU/mL with 95% probability. The simple nature of paper-based detection system with a great sensitivity and specificity would provide an effective means of evaluating the safety of food and environmental samples.A novel bifunctional oligonucleotide (OND) probe with single fluorescent group HEX labelled at 5′-end was designed for detecting trace Ag(I) and Pb(II) in real samples. In the presence of Ag(I), the hairpin structure originating from Ag(I) induced cytosine-Ag(I)-cytosine mismatches causes the proximity of the HEX to the consecutive guanine bases (G)4 at 3′-terminal, resulting in the fluorescence quenching of the HEX. While in the presence of Pb(II), the G-quadruplex structure originating from two G-quartet planes by the intramolecular hydrogen bond with Pb(II) also causes the HEX approaching the (G)4 terminal and consequently the fluorescence quenching. The results showed the quantitative detection of trace Ag(I) and Pb(II) both in the linear response ranges of 1.0-20.0 × 10-9 mol L-1 with no visible interferences of other 11 metal ions observed. And the detection limits were 82 × 10-12 mol L-1 for Ag(I), 92 × 10-12 mol L-1 for Pb(II), respectively. The fluorescence quenching mechanism of the (G)4 to HEX was verified to be the photoinduced electron transfer in the aspect of thermodynamics. This method provided a feasible application for sensitive and selective detection of Pb(II) and Ag(I) in water and Chinese traditional herbs with convenient operation.Assembly and bonding are major obstacles in manufacturing of functionally integrated fluidic devices. Here we demonstrate a single-material 3D printed device with an integrated porous structure capable of filtering particulate matter for the colourimetric detection of iron from soil and natural waters. Selecting a PolyJet 3D printer for its throughput, integrated filters were created exploiting a phenomenon occurring at the interface between the commercially available build material (Veroclear-RGD810) and water-soluble support material (SUP707). The porous properties were tuneable by varying the orientation of the print head relative to the channel and by varying the width of the build material. Porous structures ranging from 100 to 200 μm in thickness separated the sample and reagent chambers, filtering particles larger than 15 μm in diameter. Maintaining the manufacturing throughput of the Polyjet printer, 221 devices could be printed in 1.5 h (∼25 s per device). Including the 12 h post-processing soak in sodium hydroxide to remove the solid support material, the total time to print and process 221 devices was 13.5 h (3.6 min per device), with a material cost of $2.50 each. The applicability of the fluidic device for point of collection analysis was evaluated using colourimetric determination of iron from soil slurry and environmental samples. Following the reduction of Fe3+ to Fe2+ using hydroxylammonium chloride, samples were introduced to the fluidic device where particulate matter was retained by the filter, allowing for particulate-free imaging of the red complex formed with 1,10-phenanthroline using a smartphone camera. The calibration curve ranged from of 1-100 mg L-1 Fe2+ and good agreement (95%) was obtained between the point of collection device and Sector Field ICP-MS.The CRISPR/Cas12a system has displayed remarkable potential in the development of new methods for nucleic acid detection owing to the trans-cleavage activity of Cas12a. Despite the tremendous development in recent years, existing CRISPR/Cas12a-based methods have several limitations such as the time-consuming process, which takes up to 2 h, and the risk of aerosol contamination during DNA amplicon transfer. AZD9291 research buy Herein, we propose a CRISPR/Cas12a-based fluorescence detection platform named “Cas12aFDet” for rapid nucleic acid detection that overcomes these limitations. By integrating PCR or recombinase-aided amplification (RAA) methods with Cas12a-mediated cleavage in a sealed reaction tube, Cas12aFDet-based detection of amplified products could be accomplished within 15 min, while avoiding amplicon contamination. The detection limits of PCR-based Cas12aFDet and RAA-based Cas12aFDet were determined to be 3.37 × 101 cfu/mL and 1.35 × 102 cfu/mL of Listeria monocytogenes serotype 4c in pure culture, respectively. Most importantly, RAA-based Cas12aFDet exhibited 0.

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