Cheng Zhi Huang

Highly selective detection of phosphate in very complicated matrixes with an off–on fluorescent probe of europium-adjusted carbon dots

A simple method for phosphate (Pi) detection is established by developing an off-on fluorescence probe of europium-adjusted carbon dots (CDs), which has been successfully applied to the detection of Pi in very complicated matrixes such as artificial wetlands system.

One-pot hydrothermal synthesis of highly luminescent nitrogen-doped amphoteric carbon dots for bioimaging from Bombyx mori silk – natural proteins

Nitrogen-doped carbon dots (CDs) have attracted great interest due to their extraordinary properties, especially their enhanced emission efficiency, and thus a facile synthesis of nitrogen-doped CDs with high emission efficiency is critical for practical applications. To improve the emission efficiency of CDs, herein we employed Bombyx mori silk, which has high nitrogen content, as a raw material to prepare photoluminescent nitrogen-doped carbon dots through one-pot hydrothermal synthesis, and found that the as-prepared CDs have a photoluminescence (PL) quantum yield of 13.9%, and display amphoteric properties depending on the pH, are highly photostable, have low toxicity and are suitable for bioimaging.

One-Step Label-Free Optical Genosensing System for Sequence-Specific DNA Related to the Human Immunodeficiency Virus Based on the Measurements of Light Scattering Signals of Gold Nanorods

A one-step label-free optical genosensing method has been developed in this contribution by taking short DNA target with its sequence related to the human immunodeficiency virus type 1 (HIV-1) as an example. By employing anisotropic nonspherical and positively charged gold nanorods (Au-NRs) as the recognition platform, which show high stability against aggregation under high ionic strength conditions without any additional stable reagent, we found that the addition of target DNA to the mixture of nonmodified Au-NRs suspension and label-free probe DNA in high ionic strength buffer leads to a color change from red to light purple in less than 5 min, displaying strong plasmon resonance light scattering (PRLS) signals. Mechanism investigations showed that the strong PRLS signals should be ascribed to the aggregation of Au-NRs induced by the formed double-stranded oligonucleotides (dsDNA) from the hybridization of target DNA with probe DNA. With the PRLS signals, we monitored the hybridization process of a 21-mer single-stranded oligonucleotide (ssDNA) from the HIV-1 U5 long terminal repeat (LTR) sequence with its complementary oligonucleotide and detected the effect of single-base-pair mismatches. Two polymerase chain reaction (PCR) amplicon artificial samples derived from Mycobacterium tuberculosis glmS and genes encoding for Bacillus glucanase and an HIV-1 LTR sample isolated from HIV-1-positive blood were detected with satisfactory results, showing that the present method has simplicity, sensitivity, specificity, and reliability for sequence-specific DNA detection related to the HIV gene.

Carbon Nanotubes as a Low Background Signal Platform for a Molecular Aptamer Beacon on the Basis of Long-Range Resonance Energy Transfer

Although holding the advantages of both an aptamer and a molecular beacon (MB), a molecular aptamer beacon (MAB) needs complicated and expensive modifications at both of its ends and usually has a high background signal because of the low energy transfer efficiency between the donor and the acceptor. To overcome these shortcomings, in this study, we develop a long-range resonance energy transfer (LrRET) system by separating the donor from the acceptor, wherein only one end of the MAB is fluorescently labeled and acts as the energy donor and multiwalled carbon nanotubes (MWCNTs) are introduced as the energy acceptor. To test the feasibility of the newly designed MAB system, adenosine triphosphate (ATP) has been employed as a proof-of-concept target. It is found that the fluorescence of the designed MAB is completely quenched by MWCNTs, supplying a very low background signal. Then the quenched fluorescence is recovered significantly with the addition of ATP, so that ATP can be detected in the range of 0.8-80 μM with a limit of detection of 0.5 μM (3σ). Compared with the conventional fluorescence resonance energy transfer, the efficiency of LrRET between the dye and MWCNTs is much higher. Since only one end of the MAB needs the modification, the present strategy is simple and cost-effective. Furthermore, the use of MWCNTs can greatly reduce the fluorescence background of the MAB and supply a high sensitivity, showing its generality for detection of a variety of targets.

End-to-end assembly of gold nanorods by means of oligonucleotide–mercury(II) molecular recognition

We report a novel, simple, highly selective and versatile approach for the end-to-end assembly of gold nanorods (GNRs) by means of the specific molecular recognition between thymine-rich (T-rich) oligonucleotides and mercury(II).

Rapid and selective detection of cysteine based on its induced aggregates of cetyltrimethylammonium bromide capped gold nanoparticles

A detection method of cysteine is reported in this contribution with water-soluble positively charged gold nanoparticles (Au-NPs) that were prepared by seed-mediated method and capped with cetyltrimethylammonium bromide (CTAB). In aqueous medium of pH 4.2, the CTAB-capped Au-NPs display greatly different features from those of generally prepared citrate-coated Au-NPs. It was found that in a medium of high salt concentration, the presence of cysteine could induce aggregation of CTAB-capped Au-NPs, while citrate-coated Au-NPs could get aggregation soon even if without the presence of cysteine. The cysteine-induced aggregates of CTAB-capped Au-NPs display strong plasmon resonance light scattering (PRLS) signals characterized at 566.0nm when excited by a light beam, and the PRLS intensities of the aggregates are in proportion to the concentration of cysteine in the range of 0.01-0.40microgmL(-1) with the limit of detection (3sigma) being 2.9ngmL(-1). No amino acids in the samples interfere with the detection, and cysteine in artificial samples could be detected with the recovery between 95.3% and 105.9%, and R.S.D. is less than 3.6%.

Aptamer-Based Silver Nanoparticles Used for Intracellular Protein Imaging and Single Nanoparticle Spectral Analysis

Aptamer-adapted silver nanoparticles (Apt-AgNPs) were developed as a novel optical probe for simultaneous intracellular protein imaging and single nanoparticle spectral analysis, wherein AgNPs act as an illuminophore and the aptamer as a biomolecule specific recognition unit, respectively. It was found that streptavidin-conjugated and aptamer-functionalized AgNPs show satisfactory biocompatibility and stability in cell culture medium, and thus not only can act as a high contrast imaging agent for both dark-field light scattering microscope and TEM imaging but also can inspire supersensitive single nanoparticle spectra for potential intercellular microenvironment analysis. Further investigations showed that caveolae-related endocytosis is likely a necessary pathway for Apt-AgNPs labeled PrP(c) internalization in human bone marrow neuroblastoma cells (SK-N-SH cells). The integrated capability of Apt-AgNPs to be used as light scattering and TEM imaging agents, along with their potential use for single nanoparticle spectral analysis, makes them a great promise for future biomedical imaging and disease diagnosis.

A gold nanoparticles-based colorimetric assay for alkaline phosphatase detection with tunable dynamic range

In this report, a simple and label-free colorimetric assay was developed for detecting alkaline phosphatase (ALP). Based on the conjugated gold nanoparticle/adenosine triphosphate (AuNP/ATP) sensing system, this assay is highly sensitive and selective. In this system, ATP induces the aggregation of cetyltrimethylammonium bromide (CTAB)-capped AuNPs and ALP stimulates the disaggregation of AuNPs by converting ATP into adenosine through an enzymatic dephosphorylation reaction. Hence, the presence of ALP can be visually observed (gray-to-red color change) and monitored by the shift of the surface plasmon resonance (SPR) absorption band of AuNPs. Furthermore, the dynamic range of the method can be varied by addition of different metal ions (e.g. 100-600unit/L to 5.0-100unit/L and 0.2-20unit/L in the presence of Ca(2+) and Pb(2+), respectively). The feasibility of this sensitive and specific assay with a tunable dynamic range was demonstrated to be consistent even in human serum samples.

A visual detection of hydrogen peroxide on the basis of Fenton reaction with gold nanoparticles.

Nowadays, hydrogen peroxide (H(2)O(2)) has attracted more and more attentions in biochemical fields owing to its important role in biological systems. In this contribution, we propose a novel assay for the detection of H(2)O(2) based on the cleavage of ssDNA on gold nanoparticles (AuNPs). It was known that AuNPs could be stable in the presence of single-stranded DNA (ssDNA) which prevents the salt-induced aggregation of AuNPs in solution owing to the electrostatic repulsion. However, hydroxyl radical (HO*) generated from Fenton reaction could cleave the ssDNA and induce the aggregation of AuNPs. Therefore, color change from red to blue owing to the plasmon resonance absorption (PRA) of AuNPs can be observed by the naked eyes and enhancement of plasmon resonance light scattering could be measured with a common spectrofluorometer. The values of A(650)/A(520) of the PRA band were found to be linearly proportional to the concentration of H(2)O(2) in the range of 2.0 x 10(-7) to 8.0 x 10(-6) mol L(-1) with the limit of determination (LOD) being 40 nmol L(-1) (S/N=3), and thus the content of H(2)O(2) in rat encephalon extraction could be successfully detected with the recovery in the range of 98-103%.

Sensitive Discrimination and Detection of Prion Disease-Associated Isoform with a Dual-Aptamer Strategy by Developing a Sandwich Structure of Magnetic Microparticles and Quantum Dots

The major challenge of prion disease diagnosis at the presymptomatic stage is how to sensitively or selectively discriminate and detect the minute quantity of disease-associated prion protein isoform (PrP(Res)) in complex biological systems such as serum and brain homogenate. In this contribution, we developed a dual-aptamer strategy by taking the advantages of aptamers, the excellent separation ability of magnetic microparticles (MMPs), and the high fluorescence emission features of quantum dots (QDs). Two aptamers (Apt1 and Apt2), which can recognize their two corresponding distinct epitopes of prion proteins (PrP), were coupled to the surfaces of MMPs and QDs, respectively, to make MMPs-Apt1 and QDs-Apt2 ready at first, which then could be coassociated together through the specific recognitions of the two aptamers with their two corresponding distinct epitopes of PrP, forming a sandwich structure of MMPs-Apt1-PrP-Apt2-QDs and displaying the strong fluorescence of QDs. Owing to the different binding affinities of the two aptamers with PrP(Res) and cellular prion protein (PrP(C)), both of which have distinct denaturing detergent resistance, our dual-aptamer strategy could be applied to discriminate PrP(Res) and PrP(C) successfully in serum. Further identifications showed that the present dual-aptamer assay could be successfully applied to the detection of PrP in 0.01% brain homogenate, about 1000-fold lower than that of commonly applied antibody-mediated assays, which can detect PrP just in 10% brain homogenate, indicating that the present designed dual-aptamer assay is highly sensitive and adequate for clinical diagnosis without isolation of target protein prior to assay.