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Conventional photoelectrochemical (PEC) analysis mostly utilizes photoactive material modified planar indium tin oxides (ITOs) to obtain photocurrent responses for the measurement of analytes in solution. In this work, a CdS quantum dot (QD) modified nanopipette was prepared for the PEC analysis of the alkaline phosphatase (ALP) activity in single MCF-7 cells. The nanopipette was filled with ascorbic acid 2-phosphate (AAP) that was egressed outside the nanopipette by electrochemical pumping. Next, AAP was catalyzed by ALP to generate ascorbic acid (AA), which is an efficient electron donor for CdS QDs under illumination. Based on the result that the nanopipette showed a linear photocurrent response to AA, a nearly linear correlation between the photocurrent and the activity of ALP was established. Accordingly, using these CdS QD modified nanopipettes, the ALP activity in single MCF-7 cells was determined to be 0.12 U mL−1 by PEC analysis. This work does not expand the application of PEC bioanalysis, but offers a new strategy for single cell analysis. [ABSTRACT FROM AUTHOR]

Drop-on-demand microkits with a diameter of ∼20 μm are used to measure the activity of acetylcholinesterase (AChE) in a brain slice with single-cell resolution. The relative standard deviation from 25 cellular regions reached 73.3% exhibiting the difference of enzyme activity in the brain slice. Therefore, this approach utilizing the well-established kits provides an alternative single-cell-resolved strategy for the elucidation of enzymatic heterogeneity at the tissue level. [ABSTRACT FROM AUTHOR]

Potassium-selective polymer dots (K-Pdots) containing potassium ionophores were for the first time used for photoelectrochemical (PEC) analysis and yielded sensitive and specific detection of potassium ions. The successful PEC analysis using ion-selective Pdots underscored the effectiveness of the strategy deployed and suggested the potential universality of this strategy for the detection of metal ions, which should advance the development of PEC sensors in ion analysis. [ABSTRACT FROM AUTHOR]

Upconverting ion-selective nanoparticles that emit light at the near-infrared region are prepared here. The transport of calcium ions induces the deprotonation of the incorporated chromoionophore (P6) through ion exchange resulting in an increase in the emission of UCNPs for the detection of intracellular calcium ions. [ABSTRACT FROM AUTHOR]

The molecular profiling of single axons and dendrites in living neurons could provide important information for the better understanding of neuron function. Here, electrosyringe-assisted electrospray mass spectrometry (MS) is established for the first time to achieve intracellular sampling from one axon or dendrite in living neurons for mass spectrometric analysis. The key is the insertion of a ∼130 nm capillary tip into one axon or dendrite to load the cytosol through electro-osmotic flow. The ionization efficiency from the nano-capillary is enhanced to guarantee mass spectrometric analysis of multiple components from the axon and dendrite. Higher levels of pyroglutamic acid and glutamic acid are revealed in the axon compared to in the body and dendrite. This uneven distribution is in accordance with the accumulation of neurotransmitters in the axon for information delivery. The achievement of electrosyringe-assisted electrospray MS is to unveil the molecular distribution in the whole living neuron, which offers the feasibility to deeply investigate molecular communication between the axon/dendrite and the body inside neurons. [ABSTRACT FROM AUTHOR]

Proximity induced intramolecular nucleotide strand displacement, which can be simply performed in a single tube or in a complex cellular environment, is one of the key mechanisms for the detection of biological targets, especially for significant genetic molecules. The host factor for RNA phage Qb replication (Hfq), with two distinct single stranded RNA binding sites, has excellent properties as an affinity ligand in a proximity induced reaction. In this research, a versatile RNA chaperone-Hfq assisted RNA annealing strategy for the sensitive detection of the intermediate product, oligouridylated RNA, in a genetic regulation process was developed. Benefiting from the high binding affinity of Hfq for the probe and the target, the sensitive determination of oligouridylated RNA in cell lysis and human cervical cancer (HeLa) cells was successfully achieved. This study has also revealed that the Hfq assisted RNA annealing strategy can be further extended and applied in specific microRNA analysis, and RNA related tumorigenicity and disease diagnosis. [ABSTRACT FROM AUTHOR]

A novel strategy for the preparation of an aptamer based organic–inorganic hybrid affinity monolithic column was developed successfully using gold nanoparticles (GNPs) as an intermediary for a sandwich structure to realize the functional modification of the surface of the monolithic matrix. This monolithic matrix was facilely pre-synthesized via one-step co-condensation. Due to the high surface-to-volume ratio of GNPs and the large specific surface area of the hybrid matrix, the average coverage density of aptamers on the hybrid monolith reached 342 pmol μL−1. With the combination of an aptamer based hybrid affinity monolithic column and enzymatic chromogenic assay, the quantitation and detection limits of thrombin were as low as 5 nM and 2 nM, respectively. These results indicated that the GNPs attached monolith provided a novel technique to immobilize aptamers on an organic–inorganic hybrid monolith and it could be used to achieve highly selective recognition and determination of trace proteins. [ABSTRACT FROM AUTHOR]

Depending on the situation, metal ions may either play beneficial roles or be harmful to human health and ecosystems. Sensitive and accurate detection of metal ions is thus a critical issue in the field of analytical sciences and great efforts have been devoted to the development of various metal ion sensors. Photoelectrochemical (PEC) detection is an emerging technique for the bio/chemical detection of metal ions, and features a fast response, low cost and high sensitivity. Using representative examples, this review will first introduce the fundamentals and summarize recent progress in the PEC detection of metal ions. In addition, interesting strategies for the design of particular PEC metal ion sensors are discussed. Challenges and opportunities in this field are also presented. [ABSTRACT FROM AUTHOR]

A novel three-dimensionally structured DNA probe is reported to realize in situ“off–on” imaging of intracellular telomerase activity. The probe consists of a DNA tetrahedron and a hairpin DNA on one of the vertices of the DNA tetrahedron. It is composed of four modified DNA segments: S1–Au nanoparticle (NP) inserting a telomerase strand primer (TSP) and S2–S4, three Cy5 dye modified DNA segments. Fluorescence of Cy5 at three vertices of the DNA tetrahedron is quenched by the Au NP at the other vertex due to the effective fluorescence resonance energy transfer (FRET) (“off” state). When the probe meets telomerase, the hairpin structure changes to rod-like through complementary hybridization with the telomerase-triggered stem elongation product, resulting in a large distance between the Au NP and Cy5 and the recovery of Cy5 fluorescence (“on” state). The molar ratio of 3 : 1 between the reporter (Cy5) and the target related TSP makes the probe show high sensitivity and recovery efficiency of Cy5 in the presence of telomerase extracted from HeLa cells. Given the functional and compact nanostructure, the mechanically stable and noncytotoxic nature of the DNA tetrahedron, this FRET-based probe provides more opportunities for biosensing, molecular imaging and drug delivery. [ABSTRACT FROM AUTHOR]

Proximity induced intramolecular nucleotide strand displacement, which can be simply performed in a single tube or in a complex cellular environment, is one of the key mechanisms for the detection of biological targets, especially for significant genetic molecules. The host factor for RNA phage Qb replication (Hfq), with two distinct single stranded RNA binding sites, has excellent properties as an affinity ligand in a proximity induced reaction. In this research, a versatile RNA chaperone-Hfq assisted RNA annealing strategy for the sensitive detection of the intermediate product, oligouridylated RNA, in a genetic regulation process was developed. Benefiting from the high binding affinity of Hfq for the probe and the target, the sensitive determination of oligouridylated RNA in cell lysis and human cervical cancer (HeLa) cells was successfully achieved. This study has also revealed that the Hfq assisted RNA annealing strategy can be further extended and applied in specific microRNA analysis, and RNA related tumorigenicity and disease diagnosis.

A novel three-dimensionally structured DNA probe is reported to realize in situ"off-on" imaging of intracellular telomerase activity. The probe consists of a DNA tetrahedron and a hairpin DNA on one of the vertices of the DNA tetrahedron. It is composed of four modified DNA segments: S1-Au nanoparticle (NP) inserting a telomerase strand primer (TSP) and S2-S4, three Cy5 dye modified DNA segments. Fluorescence of Cy5 at three vertices of the DNA tetrahedron is quenched by the Au NP at the other vertex due to the effective fluorescence resonance energy transfer (FRET) ("off" state). When the probe meets telomerase, the hairpin structure changes to rod-like through complementary hybridization with the telomerase-triggered stem elongation product, resulting in a large distance between the Au NP and Cy5 and the recovery of Cy5 fluorescence ("on" state). The molar ratio of 3 : 1 between the reporter (Cy5) and the target related TSP makes the probe show high sensitivity and recovery efficiency of Cy5 in the presence of telomerase extracted from HeLa cells. Given the functional and compact nanostructure, the mechanically stable and noncytotoxic nature of the DNA tetrahedron, this FRET-based probe provides more opportunities for biosensing, molecular imaging and drug delivery.