A water-soluble fluorescence resonance energy transfer (FRET) probe for hypochlorous acid (HOCl), dansyl rhodamine B piperazinoacetohydrazide, was designed, synthesized and characterized. The dansyl moiety in the probe acted as a FRET donor and the rhodamine moiety acted as a FRET acceptor. The two moieties were connected by a HOCl-cleavable active bond, and cleavage of this linker decreased the FRET efficiency and increased the fluorescence intensity of the donor at 501 nm. The water solubility of the probe was improved compared with other probes by introduction of the cationic rhodamine fluorophore. As a result, the probe could be used to detect HOCl in aqueous biosystems with a linear range of 2-10 mol/L and a detection limit of 80 nmol/L (signal- to-noise = 3). The probe was successfully applied to fluorescence imaging of HOCl in HeLa cells.
This study demonstrates an on-line method for continuous measurements of cerebral hypoxanthine in the freely moving rats with integration of selective electrochemical biosensing with in vivo microdialysis sampling. The selective electrochemical biosensing is achieved by using xanthine oxidase (XOD) as the specific sensing element and Prussian blue (PB) as the electrocatalyst for the reduction of H2O2 generated from the oxidase-catalyzed reaction. The method is virtually interference-free from the co-existing electroactive species in the brain and exhibits a good stability and reproducibility. Upon integrated with in vivo microdialysis, the on-line method is well suitable for continuous measurements of cerebral hypoxanthine of freely moving rats, which is illustrated by the measurements of the microdi-alysates after the hypoxanthine standard was externally infused into the rat brain. This study essentially offers a facile on-line electrochemical approach to continuous measurements of cerebral hypoxanthine and could find some interesting applications in physiological and pathological investigations associated with hypoxanthine.
Vertigo is one of the most common clinical symptoms. However, the chemical processes involved in the pathological mechanism of vertigo remain to be fully understood. In this study, we investigate the dynamic changes in the magnesium (Mg 2+ ) concentration in medial vestibular nucleus (MVN) of guinea pigs following vertigo induced by vestibular ice water stimulation with an electrochemical detection method consisting of in vivo microdialysis and on-line selective electrochemical detection. Electrochemical detection of Mg 2+ was accomplished based on the current enhancement of Mg 2+ towards the electrocatalytic oxidation of NADH at the electrodes modified with the polymerized film of toluidine blue O (TBO). Selectivity for the on-line electrochemical detection against Ca 2+ was achieved by using ethyleneglcol-bis(2-aminoethylether) tetraacetic acid (EGTA) as the selective masking agent for Ca 2+ . The basal level of the extracellular Mg 2+ in the MVN of guinea pigs was determined to be 759.7 ± 176.2 M(n = 16). Upon ice water irrigation of the left external ear canal, the concentration of Mg 2+ in the MVN decreases significantly, reaches 72 ± 6% (n = 8) of the basal level, and maintains for at least 1000 s. Control experiments reveal that neither warm water irrigation of the external ear canal nor ice water irrigation of the auricle induces the decrease in the concentration of Mg 2+ in the MVN. These results demonstrate that the extracellular Mg 2+ in the MVN decreases significantly following vertigo induced by vestibular ice water stimulation. This demonstration suggests that Mg 2+ might play an important role in the pathological mechanism of vertigo.
Immunoassays are useful for many bioassays. Many new techniques and materials are introduced into the immunoassay to improve the efficiency. This paper reviews recent progress in the application of microfluidic systems and gold nanoparticles in immunoassay. The micro/nano technologies and materials can offer good sensitivity, fast detection, cost-effectiveness and easy signal readout. In particular, the miniaturization of microfluidics and colorimetric assays based on gold nanoparticles have dramatically improved the efficiency of immunoassays.
This review begins with an overview of the appealing properties and various applications of gold nanoparticles, and briefly summarizes recent advances in using unmodified gold nanoparticles to detect different kinds of targets including nucleic acids, proteins, metal ions and small organic molecules. The key point to the unmodified gold nanoparticle-based visual detection assay is to control dispersion and aggregation of colloidal nanoparticles by targets of interest, which usually relies on affinities between gold nanoparticles and targets. The degree of dispersion or aggregation can be visualized through the change of the solution color or the precipitation of nanoparticles from the solution. Thus, the existence of the target molecules can be trans-lated into optical signals and monitored by the naked eye conveniently. Finally, some future prospects of this research field are given.
