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A novel strategy for the enhancement of electrochemiluminescence (ECL) was developed by combining CdSe nanocrystals (NCs), carbon nanotube-chitosan (CNT-CHIT), and 3-aminopropyl-triethoxysilane (APS). A label-free ECL immunosensor for the sensitive detection of human IgG (HIgG) was fabricated. The colloidal solution containing CdSe NCs/CNT-CHIT composite was first covered on the Au electrode surface to form a robust film, which showed high ECL intensity and good biocompatibility. After APS as a cross-linker was covalently conjugated to the CdSe NCs/CNT-CHIT film, the ECL intensity was greatly enhanced. And, an intensity about 20-fold higher than that of the CdSe NCs/CNT-CHIT film was observed. After antibody was bound to the functionalized film via glutaric dialdehyde (GLD), the modified electrode could be used as an ECL immunosensor for the detection of HIgG. The specific immunoreaction between HIgG and antibody resulted in the decrease in ECL intensity. The ECL intensity decreased linearly with HIgG concentration in the range of 0.02-200 ng mL(-1), and the detection limit was 0.001 ng mL(-1). The immunosensor has the advantages of high sensitivity, speed, specificity, and stability and could become a promising technique for protein detection.

Summary Recent advances in nanoscale science and technology have generated nanomaterials with unique optical properties. Over the past decade, numerous fluorescent nanoprobes have been developed for highly sensitive and selective sensing and imaging of metal ions, both in vitro and in vivo. In this review, we provide an overview of the recent development of the design and optical properties of the different classes of fluorescent nanoprobes based on noble metal nanomaterials, upconversion nanoparticles, semiconductor quantum dots, and carbon-based nanomaterials. We further detail their application in the detection and quantification of metal ions for environmental monitoring, food safety, medical diagnostics, as well as their use in biomedical imaging in living cells and animals.

CONSPECTUS: Metal ions can be beneficial or toxic depending on their identity, oxidation state, and concentration. Therefore, the ability to detect and quantify different types of metal ions using portable sensors or in situ imaging agents is important for better environmental monitoring, in vitro medical diagnostics, and imaging of biological systems. While numerous metal ions in different oxidation states are present in the environment and biological systems, only a limited number of them can be detected effectively using current methods. In this Account, we summarize research results from our group that overcome this limitation by the development of a novel class of activity-based sensors based on metal-dependent DNAzymes, which are DNA molecules with enzymatic activity. First, we have developed an in vitro selection method to obtain DNAzymes from a large DNA library of up to 10 15 sequences that can carry out cleavage of an oligonucleotide substrate only in the presence of a specific metal ion with high selectivity. Negative selection steps can further be used to improve the selectivity against potentially competing targets by removing sequences that recognize the competing metal ions. Second, we have developed a patented catalytic beacon method to transform the metal-dependent DNAzyme cleavage reaction into a turn-on fluorescent signal by attaching a fluorophore and quenchers to the DNAzyme complex. Because of the difference in the melting temperatures of DNA hybridization before and after metal-ion-dependent cleavage of the DNAzyme substrate, the fluorophore on the DNA cleavage product can be released from its quenchers to create a turn-on fluorescent signal. Because DNAzymes are easy to conjugate with other signaling moieties, such as gold nanoparticles, lanthanide-doped upconversion nanoparticles, electrochemical agents, and gadolinium complexes, these DNAzymes can also readily be converted into colorimetric sensors, upconversion luminescence sensors, electrochemical sensors, or magnetic resonance contrast agents. In addition to describing recent progress in developing and applying these metal ion sensors for environmental monitoring, point-of-care diagnostics, cellular imaging, and in vivo imaging in zebrafish, we summarize major advantages of this class of activity-based sensors. In addition to advantages common to most activity-based sensors, such as enzymatic turnovers that allow for signal amplification and the use of initial rates instead of absolute signals for quantification to avoid interferences from sample matrices, the DNAzyme-based sensors allow for in vitro selection to expand the method to almost any metal ion under a variety of conditions, negative selection to improve the selectivity against competing targets, and reselection of DNAzymes and combination of active and inactive variants to fine-tune the dynamic range of detection. The use of melting temperature differences to separate target binding from signaling moieties in the catalytic beacon method allows the use of different fluoropho...

Gelatin-stabilized gold nanoparticles (AuNPs-gelatin) with hydrophilic and biocompatible were prepared with a simple and "green" route by reducing in situ tetrachloroauric acid in gelatin. The nanoparticles showed the excellent colloidal stability. UV-vis spectra, transmission electron microscopy (TEM), and atomic force microscopy revealed the formation of well-dispersed AuNPs with different sizes. By combination of the biocompatibility of AuNPs and excellent conductivity of carboxylic single-walled carbon nanotubes (c-SWNTs), a novel nanocomposite was designed for the immobilization and cytosensing of HL-60 cells at electrodes. The immobilized cells showed sensitive voltammetric response, good activity, and increased electron-transfer resistance. It can be used as a highly sensitive impedance sensor for HL-60 cells ranging from 1 x 10(4) to 1 x 10(7) cell mL(-1) with a limit of detection of 5 x 10(3) cell mL(-1). Moreover, the nanocomposite could effectively facilitate the interaction of adriamycin (ADR) with HL-60 cells and remarkably enhance the permeation and drug uptake of anticancer agents in the cancer cells, which could readily lead to the induction of the cell death of leukemia cells.