Bifunctional Fe3O4@Ag nanoparticles with both superparamagnetic and antibacterial properties were prepared by reducing silver nitrate on the surface of Fe3O4 nanoparticles using the water-in-oil microemulsion method. Formation of well-dispersed nanoparticles with sizes of 60 ± 20 nm was confirmed by transmission electron microscopy and dynamic light scattering. X-ray diffraction patterns and UV–visible spectroscopy indicated that both Fe3O4 and silver are present in the same particle. The superparamagnetism of Fe3O4@Ag nanoparticles was confirmed with a vibrating sample magnetometer. Their antibacterial activity was evaluated by means of minimum inhibitory concentration value, flow cytometry, and antibacterial rate assays. The results showed that Fe3O4@Ag nanoparticles presented good antibacterial performance against Escherichia coli (gram-negative bacteria), Staphylococcus epidermidis (gram-positive bacteria) and Bacillus subtilis (spore bacteria). Furthermore, Fe3O4@Ag nanoparticles can be easily removed from water by using a magnetic field to avoid contamination of surroundings. Reclaimed Fe3O4@Ag nanoparticles can still have antibacterial capability and can be reused.
A new class of intracellular nanoprobe, termed fluorescence resonance energy transfer (FRET) nanoflares, was developed to sense mRNA in living cells. It consists of a gold nanoparticle (AuNP), recognition sequences, and flares. Briefly, the AuNP functionalized with recognition sequences hybridized to flares, which are designed as hairpin structures and fluorescently labeled donors and acceptors at two ends, respectively. In the absence of targets, the flares are captured by binding with the recognition sequences, separating of the donor and acceptor, and inducing low FRET efficiency. However, in the presence of targets, the flares are gradually displaced from the recognition sequences by the targets, subsequently forming hairpin structures that bring the donor and acceptor into close proximity and result in high FRET efficiency. Compared to the conventional single-dye nanoflares, the upgraded FRET nanoflares can avoid false positive signals by chemical interferences (such as nuclease and GSH) and thermodynamic fluctuations. Moreover, the signal generation in FRET nanoflares can be easily made with ratiometric measurement, minimizing the effect of system fluctuations.
China Scholarship Council (CSC); ACS; US NIH; China NSFC[20805038]; National Basic Research Program of China[2007CB935603, 2010CB732402]; China National Grand Program on Key Infectious Disease[2009ZX10004-312]; Key Project of Natural Science Foundation of China[90606003]; International Science & Technology Cooperation Program of China[2010DFB30300]; Hunan Provincial Natural Science Foundation of China[10JJ7002
Aptamers have emerged as promising molecular probes for in vivo cancer imaging, but the reported "always-on" aptamer probes remain problematic because of high background and limited contrast. To address this problem, we designed an activatable aptamer probe (AAP) targeting membrane proteins of living cancer cells and achieved contrast-enhanced cancer visualization inside mice. The AAP displayed a quenched fluorescence in its free state and underwent a conformational alteration upon binding to target cancer cells with an activated fluorescence. As proof of concept, in vitro analysis and in vivo imaging of CCRF-CEM cancer cells were performed by using the specific aptamer, sgc8, as a demonstration. It was confirmed that the AAP could be specifically activated by target cancer cells with a dramatic fluorescence enhancement and exhibit improved sensitivity for CCRF-CEM cell analysis with the cell number of 118 detected in 200 μl binding buffer. In vivo studies demonstrated that activated fluorescence signals were obviously achieved in the CCRF-CEM tumor sites in mice. Compared to always-on aptamer probes, the AAP could substantially minimize the background signal originating from nontarget tissues, thus resulting in significantly enhanced image contrast and shortened diagnosis time to 15 min. Furthermore, because of the specific affinity of sgc8 to target cancer cells, the AAP also showed desirable specificity in differentiating CCRF-CEM tumors from Ramos tumors and nontumor areas. The design concept can be widely adapted to other cancer cell-specific aptamer probes for in vivo molecular imaging of cancer.switchable aptamer probe | in vivo imaging | activatable fluorescent molecular imaging | cancer detection | cell surface protein
A novel, high sensitive, and specific DNA assay based on gold nanoparticle (AuNP) colorimetric detection and hybridization chain reaction (HCR) amplification has been demonstrated in this article. Two hairpin auxiliary probes were designed with single-stranded DNA (ssDNA) sticky ends which stabilize AuNPs and effectively prevent them from salt-induced aggregation. The target DNA hybridized with the hairpin auxiliary probes and triggered the formation of extended double-stranded DNA (dsDNA) polymers through HCR. As a result, the formed dsDNA polymers provide less stabilization without ssDNA sticky ends, and AuNPs undergo aggregation when salt concentration is increased. Subsequently, a pale purple-to-blue color variation is observed in the colloid solution. The system is simple in design and convenient in operation. The novel strategy eliminates the need for enzymatic reactions, separation processes, chemical modifications, and sophisticated instrumentation. The detection and discrimination process can be seen with the naked eye. The detection limit of this method is lower than or at least comparable to previous AuNP-based methods. Importantly, the protocol offers high selectivity for the determination between perfectly matched target oligonucleotides and targets with single base-pair mismatches.
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