Electrochemically Triggered Release of Reagent to the Proximal Leaflet of a Microcavity Supported Lipid Bilayer.

A novel and versatile approach to electrichemically triggering the release of a reagent, ß-cyclodextrin (ß-CD), selectively to the proximal leaflet of a supported lipid bilayer is described. Selective delivery is achieved by creating a spanning lipid bilayer across a microcavity array and exploiting the irreversible redox disassembly of the host-guest complex formed between thiolated ferrocene (Fc) and ß-cyclodextrin (ß-CD) in the presence of chloride. Self-assembled monolayers of the ferrocene-alkanethiols were formed regioselectively on the interior surface of highly ordered 2.8 µm cavities while the exterior top surface of the array was blocked with a monolayer of mercaptoethanol. The Fc monolayers were complexed with ß-CD or ß-CD-conjugated to streptavidin (ß-CD-SA). Phospholipid bilayers were then assembled across the array via combined Langmuir-Blodgett/vesicle fusion leading to a spanning bilayer suspended across the aqueous filled microcavities. Upon application of a positive potential, ferrocene is oxidized to ferrocinium cation, disrupting the inclusion complex and leading to the release of the ß-CD into the microcavity solution where it diffuses to the lower leaflet of the suspended bilayer. Disassembly of the supramolecular complex within the cavities and binding of the ß-CD-SA to a biotinylated bilayer was followed by voltammetry and impedance spectroscopy where it caused a large increase in membrane resistance. For unmodified ß-CD, the extraction of cholesterol from a cholesterol containing bilayer was evident in a decrease in the bilayer resistance. For the first time, this direct approach to targeted delivery of a reagent to the proximal layer of a lipid bilayer offers the potential to build models of bidirectional signaling (inside-out vs outside-in) in cell membrane model systems.

Magnetic and noble metal nanocomposites for separation and optical detection of biological species.

Nanoalloys and nanocomposites are widely studied classes of nanomaterials within the context of biological systems. They are of immense interest because of the possibility of tuning the optical, magnetic, electronic and chemical properties through particle composition and internal architecture. In principle these properties can therefore be optimized for application in biological detections such as of DNA sequences, bacteria, viruses, antibodies, antigens, and cancer cells. This article presents an overview of methods currently used for nanoalloy and nanocomposite synthesis and characterisation, focusing on Au-Ag and FexOy@Au structures as primary components in detection platforms for plasmonic and magnetically enabled plasmonic bio-sensing.

The application of water soluble, mega-Stokes-shifted BODIPY fluorophores to cell and tissue imaging.

BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) fluorophores are widely used in bioimaging to label proteins, lipids and nucleotides, but in spite of their attractive optical properties they tend to be prone to self-quenching because of their notably small Stokes shift. Herein, we compare two BODIPY compounds from a recently developed family of naphthyridine substituted BODIPY derivatives, one a visible emitting derivative (BODIPY-VIS) and one a near-infrared emitting fluorophore with a Stokes shift of approximately 165 nm as contrast reagents for live mammalian cells and murine brain tissue. The compounds were rendered water soluble by their conjugation to polyethylene glycol (PEG). Both PEGylated compounds exhibited good cell uptake compared with their parent compounds and confocal fluorescence microscopy revealed all dyes explored to be nuclear excluding, localizing predominantly within the lipophilic organelles; the endoplasmic reticulum and mitochondria. Cytotoxicity studies revealed that these BODIPY derivatives are modestly cytotoxic at concentrations exceeding 10 µM where they induce apoptosis and necrosis. Although the quantum yield of emission of the visible emitting fluorophore was over an order of magnitude greater than the Mega-Stokes shifted probe, the latter showed considerably reduced tendency to self quench and less interference from autofluorescence. The near-infrared probe also showed good penetrability and staining in live tissue samples. In the latter case similar tendency to exclude the nucleus and to localize in the mitochondria and endoplasmic reticulum was observed as in live cells. This to our knowledge is the first demonstration of such a Mega-Stokes BODIPY probe applied to cell and tissue imaging.

Synthesis, tailoring and characterization of silica nanoparticles containing a highly stable ruthenium complex.

This paper describes the synthesis and characterization of sol-gel silica nanoparticles (NPs) derived from tetraethoxysilane (TEOS) and from tetraethoxysilane and methyltriethoxysilane (TEOS-MTEOS) in which is encapsulated, an in-house synthesized, stable oxygen-sensitive ruthenium complex, ruthenium (II) (bis-2,2-bipyridyl)-2(4-carboxylphenyl) imidazo[4,5-f][1,10]phenanthroline. These NPs were characterized using dynamic light scattering, transmission electron microscopy, scanning electron microscopy, Fourier transform infrared spectroscopy and Brunauer-Emmett-Teller analysis. The spherical, stable and monodispersed NPs have been prepared using the Stöber method. It was found that the addition of prehydrolyzed MTEOS-based sol prepared in an acidic environment to the reaction mixture containing TEOS NPs synthesized for 6 h produced material with increased porosity when compared to pure silica NPs. Oxygen sensitivity, stability, photobleaching and leaching have been characterized. The hybrid NPs exhibit enhanced O2 sensitivity but a high degree of leaching when compared to pure silica NPs, which have minimum O2 sensitivity and no leaching.

Chemical and photoluminescence properties of purified poly(2-methoxyaniline-5-sulfonic acid) and oligomer.

Both the chemical and the electrochemical synthesis of poly(2-methoxyaniline-5-sulfonate) (PMAS) in aqueous media have been found to give two distinct polymer fractions with molecular weights of approximately 8-10 and 2 kDa, respectively. It is now possible to isolate the pure high molecular weight (HMWT) PMAS and low molecular weight (LMWT) PMAS oligomer and to study their individual and combined photochemistry and redox chemistry. The HMWT PMAS fraction was confirmed to be an emeraldine salt by its characteristic redox and pH switching behavior, in contrast to the oligomeric LMWT PMAS, which was inert under the same conditions. Mixtures of these two fractions exhibit photoluminescence arising from the oligomeric LMWT PMAS fraction. The observed LMWT PMAS emission was modulated by the presence of the conducting HMWT PMAS emeraldine salt via a static resonant energy transfer arising from quenching at 460 nm when excited at 355 nm. The nonlinear fluorophore-quencher behavior suggests that the two PMAS fractions are strongly associated. The behavior fitted the static Perrin quenching model in which the oligomeric LMWT PMAS fluorophore is diffusionally restricted by the presence of HMWT PMAS quencher.

Electron self-exchange in the solid-state: cocrystals of hydroquinone and bipyridyl triazole.

Solid-state voltammetry, spectroscopy, and microscopy studies have been used to probe the proton and electron conductivity within a self-assembled cocrystal, HQBpt. This crystallographically defined material contains 3,5-bis(pyridin-2-yl)-1,2,4-triazole, HBpt, dimers that are pi-stacked and hydrogen bonded to 1,4-hydroquinone, H(2)Q, in a herringbone arrangement. When deposited onto platinum microelectrodes, the cocrystal exhibits a well-defined voltammetric response corresponding to oxidation of H(2)Q to the quinone, Q, across a wide range of voltammetric time scales, electrolyte compositions, and pH values. Scanning electron microscopy reveals that redox cycling in aqueous perchlorate solutions in which the pH is systematically varied from 1 to 7 triggers electrocrystallization and the extensive formation of rodlike crystals. Fast scan rate voltammetry reveals that the homogeneous charge transport diffusion coefficient, D(app), is independent of the perchlorate concentration for 0.1 < [ClO(4)(-)] < 1.0 M (pH 6.6) at 3.14 +/- 0.11 x 10(-)(9) cm(2) s(-)(1). Moreover, D(app) is independent of the perchloric acid concentration for concentrations greater than approximately 2.0 M, maintaining a value of 4.81 +/- 0.07 x 10(-)(8) cm(2) s(-)(1). The observation that D(app) is independent of the supporting electrolyte suggests that the rate-determining step for homogeneous charge transport is not the availability of charge-compensating counterions or protons, but the dynamics of electron self-exchange between H(2)Q and Q. We have used the Dahms-Ruff formalism to determine electron self-exchange rate constants which are 2.84 +/- 0.22 x 10(9) and 9.69 +/- 0.73 x 10(10) M(-)(1) s(-)(1) for pH values greater than approximately 2.0 and less than -0.3, respectively. Significantly, these values are more than 2 orders of magnitude larger that those found for benzoquinone self-exchange reactions in aqueous solution. These results indicate that hydrogen bonds play an important role in supporting rapid electron transfer. The increase in D(app) between pH 1.0 and -0.3 is associated with protonation of the HBpt moieties, which triggers a reversible change in the material's structure.