High-Affinity Ratiometric Fluorescence Probe Based on 6-Amino-2,2′-Bipyridine Scaffold for Endogenous Zn2+ and Its Application to Living Cells

Zinc is an essential trace element involved in many biological activities; however, its functions are not fully understood. To elucidate the role of endogenous labile Zn²⁺, we developed a novel ratiometric fluorescence probe, 5-(4-methoxyphenyl)-4-(methylsulfanyl)-[2,2′-bipyridin]-6-amine (6 (rBpyZ)) based on the 6-amino-2,2′-bipyridine scaffold, which acts as both the chelating agent for Zn²⁺ and the fluorescent moiety. The methoxy group acted as an electron donor, enabling the intramolecular charge transfer state of 6 (rBpyZ), and a ratiometric fluorescence response consisting of a decrease at the emission wavelength of 438 nm and a corresponding increase at the emission wavelength of 465 nm was observed. The ratiometric probe 6 (rBpyZ) exhibited a nanomolar-level dissociation constant (K_d = 0.77 nM), a large Stokes shift (139 nm), and an excellent detection limit (0.10 nM) under physiological conditions. Moreover, fluorescence imaging using A549 human lung adenocarcinoma cells revealed that 6 (rBpyZ) had good cell membrane permeability and could clearly visualize endogenous labile Zn²⁺. These results suggest that the ratiometric fluorescence probe 6 (rBpyZ) has considerable potential as a valuable tool for understanding the role of Zn²⁺ in living systems.     

Migration analysis of micro-particles in liquids using microscopically designed external fields.

The recent development of new migration methods of micro-particles in liquids using various external fields is reviewed. The combination of a laser scattering force and a photothermal effect produced photothermal-conversion laser-photophoresis. A dielectric field generated in a planer or a capillary quadrupole electrode realized dielectrophoresis. Using a micrometer-scaled magnetic field gradient, the "Magnetophoretic velocimetry" of micro-particles was invented. Furthermore, the Lorentz force generated by combining an electric field and a magnetic field was utilized for electromagnetophoresis. These new methods were overlooked and the advantages in analytical use were discussed.     

A novel small molecule fluorescent sensor for Zn2+ based on pyridine-pyridone scaffold

The development of a water-soluble and small molecular weight fluorescent probe, 3-(4-methoxyphenyl)-4-(methylsulfanyl)-6-(pyridin-2-yl)pyridin-2(1H)-one (3), for detecting Zn²⁺ based on pyridine-pyridone skeleton is reported. We observed a clear chelation enhanced fluorescence effect of 3 in the presence of Zn²⁺. Other fluorescent properties of 3 are discussed.     

Magnetophoretic evaluation of interfacial adsorption of dysprosium(III) on a single microdroplet.

Magnetophoretic velocimetry is a novel technique to measure the magnetic susceptibility of a single microparticle. This technique could be applied to study the interfacial adsorption equilibria of a paramagnetic dysprosium(III) ion with capric, lauric or stearic acid for a single 2-fluorotoluene microdroplet. The observed magnetic susceptibility of the micro-organic droplets was reciprocally proportional to its radius in each case. From the proportional constant, the interfacial concentration of Dy(III) was determined. Furthermore, the dependences of the interfacial concentration on the initial Dy(III) concentration and pH were examined in order to analyze the adsorption equilibrium. Finally, the saturated interfacial concentration and the interfacial adsorption constant at the infinite dilution of Dy(III)-laurate complex were evaluated as 4.8 x 10(-10) mol cm(-2) and 3.4 x 10(-2) dm, respectively.     

One-pot synthesis of polysubstituted pyridine derivatives using ketene dithioacetals

Polysubstituted pyridine derivatives were synthesized by the reaction of ketene dithioacetal, 3,3-bis(methylsulfanyl)methylenemalononitrile 1b, with a variety of active methylene compounds in the presence of either sodium hydroxide or potassium hydroxide as a base in DMSO. This reaction was carried out under economical one-pot reaction conditions (cheap catalyst and solvent) and solved the problem of the odor of methanethiol, commonly derived from reactions of ketene dithioacetals. This is a significant enhancement of GSC (green and sustainable chemistry) in the field of ketene dithioacetal chemistry.     

Magnetophoretic velocity of microorganic droplets adsorbed by dysprosium(III) laurate in water

By using an improved apparatus for the observation of magnetophoresis, the magnetophoretic velocity of 2-fluorotoluene droplets including lauric acid was measured in aqueous dysprosium(III) solution. The magnetophoretic velocity of pure 2-fluorotoluene droplets was proportional to the square of the radius. On the other hand, the velocity of the organic droplets including lauric acid in the dysprosium(III) solution showed a deviation from the square radius relationship, more remarkably in smaller droplets than 2 microm in radius. These results indicated that the dysprosium(III)-laurate complex was formed at the liquid-liquid interface. This study is the first report on the detection of the interfacial complex by the magnetophoresis of the microdroplet.     

Sensitive light-scattering detection–magnetophoretic acceleration mass analysis of single microparticles in an atmosphere

Optically detected magnetophoretic acceleration mass analysis of an individual micro-particle in an atmosphere has been remarkably improved in sensitivity by using a reflective microscope objective, by which forward scattered light from a particle could be effectively collected. From the light-scattering simulation, the detection limit for the radius of a micro-particle was estimated to be smaller than 0.4 μm, and about 60 times intensity enhancement was observed for a polystyrene particle with a radius of 2.8 μm. For both paramagnetic and diamagnetic micro-particles, the mass magnetic susceptibility and the relaxation time could be determined without knowing any parameters of the particles. From the relaxation time, the mass of a particle was obtained if the radius or the density of the particle was known. For a test sample silica particles were used to adsorb paramagnetic dysprosium(III), the surface concentration of dysprosium(III) on a single particle could be successfully determined by use of this method.     

Magnetophoresis and electromagnetophoresis of microparticles in liquids

The magnetic field-induced migration of particles in liquids is a highly-promising technique for the micro-separation analysis of bioparticles, such as cells and large DNA. Here, new methods that make use of magnetophoresis and electromagnetophoresis to induce the migration of microparticles in liquids are briefly reviewed. Magnetic force and Lorentz force are utilized in the new methods. Some typical examples of the use of these methods are described, and the advantages of using a superconducting magnet for them are demonstrated.     

Magnetophoretic detection of photo-induced spin transition

Magnetophoretic velocimetry detected the spin transition of a single Co-Fe Prussian Blue analogous micro-crystal in water induced by a single-shot pulse laser.     

Transcutaneous immunization by a solid-in-oil nanodispersion

We have successfully achieved transcutaneous immunization without the use of any skin pre-treatment or immune-stimulant adjuvant by applying a solid-in-oil (S/O) nanodispersion: an oil-based nanodispersion of antigens coated with hydrophobic surfactant molecules. This finding indicates that the S/O nanodispersion has great promise for effective transcutaneous vaccination.