Zhi-Xiong Xie

Ultrasmall Near-Infrared Ag2Se Quantum Dots with Tunable Fluorescence for in Vivo Imaging

A strategy is presented that involes coupling Na(2)SeO(3) reduction with the binding of silver ions and alanine in a quasi-biosystem to obtain ultrasmall, near-infrared Ag(2)Se quantum dots (QDs) with tunable fluorescence at 90 °C in aqueous solution. This strategy avoids high temperatures, high pressures, and organic solvents so that water-dispersible sub-3 nm Ag(2)Se QDs can be directly obtained. The photoluminescence of the Ag(2)Se QDs was size-dependent over a wavelength range from 700 to 820 nm, corresponding to sizes from 1.5 ± 0.4 to 2.4 ± 0.5 nm, with good monodispersity. The Ag(2)Se QDs are less cytotoxic than other nanomaterials used for similar applications. Furthermore, the NIR fluorescence of the Ag(2)Se QDs could penetrate through the abdominal cavity of a living nude mouse and could be detected on its back side, demonstrating the potential applications of these less toxic NIR Ag(2)Se QDs in bioimaging.

Near-Infrared Electrogenerated Chemiluminescence of Ultrasmall Ag2Se Quantum Dots for the Detection of Dopamine

The near-infrared (NIR) electrogenerated chemiluminescence (ECL) of water-dispersed Ag(2)Se quantum dots (QDs) with ultrasmall size was presented for the first time. The Ag(2)Se QDs have shown a strong and efficient cathodic ECL signal with K(2)S(2)O(8) as coreactant on the glassy carbon electrode (GCE) in aqueous solution. The ECL spectrum exhibited a peak at 695 nm, consistent with the peak in photoluminescence (PL) spectrum of the Ag(2)Se QDs solution, indicating that the Ag(2)Se QDs had no deep surface traps. Dopamine was chosen as a model analyte to study the potential of Ag(2)Se QDs in the ECL analytical application. The ECL signal of Ag(2)Se QDs can also be used for the detection of the dopamine concentration in the practical drug (dopamine hydrochloride injection) containing several adjuvants such as edetate disodium, sodium bisulfite, sodium chloride and so on. The Ag(2)Se QDs could be a promising candidate emitter of ECL biosensors in the future due to their fantastic features, such as ultrasmall size, low toxicity, good water solubility, and near infrared (NIR) fluorescent emission.

Uniform fluorescent nanobioprobes for pathogen detection.

Manipulating biochemical reactions in living cells to synthesize nanomaterials is an attractive strategy to realize their synthesis that cannot take place in nature. Yeast cells have been skillfully utilized to produce desired nanoparticles through spatiotemporal coupling of intracellular nonrelated biochemical reaction pathways for formation of fluorescent CdSe quantum dots. Here, we have successfully transformed Staphylococcus aureus cells into cellular beacons (fluorescing cells), all of which are highly fluorescent and photostable with perfect uniformity. Importantly, on the basis of such cells, we efficiently fabricated fluorescent nanobioprobes by a specific interaction between the protein A expressed on the S. aureus surface and the Fc fragment domain of antibodies, avoiding the use of other common methods for cell surface modifications, such as molecular covalent connection or more difficult genetic and metabolic engineering. Coupled with immunomagnetic beads, the resulting fluorescent-biotargeting bifunctional cells, i.e., biotargeting cellular beacons, can be employed as nanobioprobes for detection of viruses, bacteria, and tumor cells. With this method, H9N2 AIV can be detected specifically with a limit of 8.94 ng/mL (based on protein content). Furthermore, diverse probes for detection of different pathogens or for other biomedical applications can be easily obtained by simply changing the antibody conjugated to the cell surface.

Mechanism-Oriented Controllability of Intracellular Quantum Dots Formation: The Role of Glutathione Metabolic Pathway

Microbial cells have shown a great potential to biosynthesize inorganic nanoparticles within their orderly regulated intracellular environment. However, very little is known about the mechanism of nanoparticle biosynthesis. Therefore, it is difficult to control intracellular synthesis through the manipulation of biological processes. Here, we present a mechanism-oriented strategy for controlling the biosynthesis of fluorescent CdSe quantum dots (QDs) by means of metabolic engineering in yeast cells. Using genetic techniques, we demonstrated that the glutathione metabolic pathway controls the intracellular CdSe QD formation. Inspired from this mechanism, the controllability of CdSe QD yield was realized through engineering the glutathione metabolism in genetically modified yeast cells. The yeast cells were homogeneously transformed into more efficient cell-factories at the single-cell level, providing a specific way to direct the cellular metabolism toward CdSe QD formation. This work could provide the foundation for the future development of nanomaterial biosynthesis.

Role of DNA in Bacterial Aggregation

The role of DNA in bacterial aggregation was determined using various types of DNA and Escherichia coli, a good model for investigating the correlation between added polymer and bacterial aggregation and adsorption of polymer to bacterial surfaces. The results of the aggregation assay suggest that extracellular DNA indeed increased the aggregation percentage of E. coli, but this effect was dependent on DNA concentration and length. Moreover, DNA promoted bacterial aggregation in a type-nonspecific way. The combined results of the aggregation assay and the adsorption assay show further that the promotion of E. coli aggregation by DNA occurred along with adsorption of DNA to E. coli. Consequently, the possible mechanisms for DNA-promoted bacterial aggregation are discussed. Using fluorescent-labeled DNA, we mapped DNA within the E. coli aggregates. Subsequently, introduction of DNase I broke up the DNA-involved E. coli aggregates. These results suggest that DNA functions as a molecular bridge to promote E. coli aggregation.

CdSe/ZnS-labeled carboxymethyl chitosan as a bioprobe for live cell imaging

A simple and convenient method for the construction of CdSe/ZnS-labeled polysaccharides as bioprobes were developed, which are highly biocompatible and photostable, and have been proven to be suitable for live cell imaging.

Exploring the mechanism of competence development in Escherichia coli using quantum dots as fluorescent probes.

The mechanism of divalent Ca2+ cation induction of Escherichia coli competence is still not fully understood, though it is a common method for introducing recombinant DNA into bacterial cells in gene engineering. Quantum dots (QDs), as a new fluorescent probe of being applied in biology research, have aroused great interest. In this paper, we explored the mechanism of E. coli competence development using QDs for the first time. Results showed that water-soluble QDs of diameter 3-4 nm could go into competent cells, but could not enter noncompetent cells. This result was further confirmed using atomic force microscopy and DNA transforming experiments, suggesting that nonphysiological, high concentrations of Ca2+ enhanced the penetrability of cell membranes so that QDs, which cannot enter cells normally due to their greater diameter (3-4 nm), can do so easily into competent cells. Therefore, we believe that, at least for E. coli, the mechanism of Ca2+-induced competence development is mediated physicochemically rather than physiologically.

Direct fluorescence in situ hybridization (FISH) in Escherichia coli with a target-specific quantum dot-based molecular beacon

Quantum dots (QDs) are inorganic fluorescent nanocrystals with excellent properties such as tunable emission spectra and photo-bleaching resistance compared with organic dyes, which make them appropriate for applications in molecular beacons. In this work, quantum dot-based molecular beacons (QD-based MBs) were fabricated to specifically detect β-lactamase genes located in pUC18 which were responsible for antibiotic resistance in bacteria Escherichia coli (E. coli) DH5α. QD-based MBs were constructed by conjugating mercaptoacetic acid-quantum dots (MAA-QDs) with black hole quencher 2 (BHQ2) labeled thiol DNA vial metal-thiol bonds. Two types of molecular beacons, double-strands beacons and hairpin beacons, were observed in product characterization by gel electrophoresis. Using QD-based MBs, one-step FISH in tiny bacteria DH5α was realized for the first time. QD-based MBs retained their bioactivity when hybridizing with complementary target DNA, which showed excellent advantages of eliminating background noise caused by adsorption of non-specific bioprobes and achieving clearer focus of genes in plasmids pUC18, and capability of bacterial cell penetration and signal specificity in one-step in situ hybridization.

Visualized investigation of yeast transformation induced with Li+ and polyethylene glycol

The effects of Li(+) and polyethylene glycol (PEG) on the genetic transformation of Saccharomyces cerevisiae were investigated by using fluorescence microscopy (FM) to visualize the binding of plasmid DNA labeled with YOYO-1 to the surface of yeast cells, scanning electron microscopy (SEM) and atomic force microscopy (AFM) to image the change in surface topography of yeast cells, coupled with transformation frequency experiments. The results showed that under the same conditions, the transformation frequencies of yeast protoplasts were much higher than those of intact yeast cells. PEG was absolutely required for the binding of DNA to the surface of intact yeast cells or yeast protoplasts, and had no effect on the surface topography of intact yeast cells or yeast protoplasts. In the presence of PEG, Li(+) could greatly enhance the binding of plasmid DNA to the surface of intact yeast cells, increase their transformation frequency, and affect their surface topography. On the other hand, no effect on the DNA binding to the surface of protoplasts and no increase in the number of transformants and no surface topography changes were found upon the treatment with Li(+) to protoplasts. In the present work, the effects of Li(+) and PEG on yeast genetic transformation were directly visualized, rather than those deduced from the results of transformation frequencies. These results indicate that cell wall might be a barrier for the uptake of plasmid DNA. Li(+) could increase the permeability of yeast cell wall, then increase the exposed sites of DNA binding on intact yeast cells. The main role of PEG was to induce DNA binding to cell surface.

Controllable synthesis of PbSe nanocubes in aqueous phase using a quasi-biosystem

By coupling two biochemical processes of reduction of Na2SeO3 with detoxification of Pb2+ in a quasi-biosystem, water-dispersed PbSe nanocubes with good monodispersity were controllably synthesized with different sizes at low temperature (90 °C) under mild conditions. The crystallization mechanism and the nature of bio-molecules influenced on the crystallization process were investigated.