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A fluorescent PEBBLE nanosensor for intracellular free zinc 总被引:2,自引:0,他引:2
The development and characterisation of a fluorescent optical PEBBLE (Probe Encapsulated By Biologically Localised Embedding) nanosensor for the detection of zinc is detailed. A ratiometric sensor has been fabricated that incorporates two fluorescent dyes; one is sensitive to zinc and the other acts as a reference. The sensing components are entrapped within a polymer matrix by a microemulsion polymerisation process that produces spherical sensors that are in the size region of 20 to 200 nm. Cellular measurements are made possible by the small sensor size and the biocompatibility of the matrix. The effects of reversibility, photobleaching and leaching have been examined, as well as the selectivity towards zinc over other cellular ions such as Na+, Ca2+, K+, and Mg2+. The dynamic range of these sensors was found to be 4 to 50 microM Zn2+ with a linear range from 15 to 40 microM. The response time for the PEBBLE is less than 4 s and the sensor is reversible. In addition, the nanosensors are photostable and leaching from the matrix, determined using a novel method, is minimal. These sensors are capable of real-time inter- and intra-cellular imaging and are insensitive to interference from proteins. 相似文献
83.
Molecular Probes' Alexa Fluor dyes are generally used for biological labeling because of their ideal fluorescent properties, but here we detail Alexa Fluor 488's nanomolar sensitivity to free iron. Furthermore, the dye has been encapsulated into a polymer nanosphere by a microemulsion method, producing <100 nm particles. These nanosensors, PEBBLEs (Probe Encapsulated By Biologically Localized Embedding) have micromolar sensitivity and are non-responsive to other metal ions of biological interest. 相似文献
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Ricci F Lai RY Heeger AJ Plaxco KW Sumner JJ 《Langmuir : the ACS journal of surfaces and colloids》2007,23(12):6827-6834
E-DNA sensors, the electrochemical equivalent of molecular beacons, appear to be a promising means of detecting oligonucleotides. E-DNA sensors are comprised of a redox-modified (here, methylene blue or ferrocene) DNA stem-loop covalently attached to an interrogating electrode. Because E-DNA signaling arises due to binding-induced changes in the conformation of the stem-loop probe, it is likely sensitive to the nature of the molecular packing on the electrode surface. Here we detail the effects of probe density, target length, and other aspects of molecular crowding on the signaling properties, specificity, and response time of a model E-DNA sensor. We find that the highest signal suppression is obtained at the highest probe densities investigated, and that greater suppression is observed with longer and bulkier targets. In contrast, sensor equilibration time slows monotonically with increasing probe density, and the specificity of hybridization is not significantly affected. In addition to providing insight into the optimization of electrochemical DNA sensors, these results suggest that E-DNA signaling arises due to hybridization-linked changes in the rate, and thus efficiency, with which the redox moiety collides with the electrode and transfers electrons. 相似文献
87.
The anaerobic oxidation of methyl p-toluate by cobalt(III) in acetic acid was investigated. Observed products were 4-carbomethoxybenzaldehyde (2), 4-carbomethoxybenzoic acid (3), 4-carbomethoxybenzyl acetate (1), 4,4'-dicarbomethoxybibenzyl (6), methyl 2,4-dimethylbenzoate (8), and methyl 3,4-dimethylbenzoate (9). Deuterium isotope labeling showed that 2 was not formed from 1, but appeared to be formed directly from methyl p-toluate via 4-carbomethoxybenzyl alcohol (5). The ratio of (2 + 3) to 1 was 0.5 with [py3Co3O(OAc)5OH[PF6] and 1.0 with cobaltic acetate. Cobaltic acetate was generated in situ by the reaction of cobaltous acetate and peracetic acid. When the oxidation was carried out in the presence of chromium (0.05 equiv based on cobalt), the ratio increased dramatically and no 6 was observed. Other transition metals such as vanadium, molybdenum, and manganese had a similar effect, but were not as effective as chromium. Chromium was observed to form a mixed-metal cluster complex with cobalt. Treatment of an acetic acid solution of cobaltous acetate and methyl isonicotinate with K2CrO4 produced a solid tentatively identified as [(MIN)3Co2CrO(OAc)6][CrO4H] (MIN = methyl isonicotinate). The selectivity for the oxidation of methyl p-toluate exhibited by the mixed-metal cluster complex was similar to that observed by the addition of chromium to oxidations using [py3Co3O(OAc)5OH[PF6]. 相似文献
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The effect of the jet-to-cross-flow velocity ratio, R, on the turbulent wake and Kármán vortex shedding for a cylindrical stack of aspect ratio AR=9 was investigated in a low-speed wind tunnel using thermal anemometry. The cross-flow Reynolds number was ReD=2.3×104, the jet Reynolds number ranged from Red=7.6×103 to 4.7×104, and R was varied from 0 to 3. The stack was partially immersed in a flat-plate turbulent boundary layer, with a boundary layer thickness-to-stack-height ratio of δ/H=0.5 at the location of the stack. From the behaviour of the turbulent wake and the vortex shedding, the flow around the stack could be classified into three regimes depending on the value of R, which were the downwash (R<0.7), cross-wind-dominated (0.7R<1.5), and jet-dominated (R1.5) flow regimes. Each flow regime had a distinct structure to the mean velocity (streamwise and wall-normal directions), turbulence intensity (streamwise and wall-normal directions), and Reynolds shear stress fields, as well as the variation of the Strouhal number and the power spectrum along the stack height. 相似文献