Anionic Conjugated Polyelectrolytes for FRET-based Imaging of Cellular Membrane Potential.

We report a Förster resonance energy transfer (FRET)-based imaging ensemble for the visualization of membrane potential in living cells. A water-soluble poly(fluorene-cophenylene) conjugated polyelectrolyte (FsPFc10) serves as a FRET donor to a voltage-sensitive dye acceptor (FluoVolt™ ). We observe FRET between FsPFc10 and FluoVolt™ , where the enhancement in FRET-sensitized emission from FluoVolt™ is measured at various donor/acceptor ratios. At a donor/acceptor ratio of 1, the excitation of FluoVolt™ in a FRET configuration results in a three-fold enhancement in its fluorescence emission (compared to when it is excited directly). FsPFc10 efficiently labels the plasma membrane of HEK 293T/17 cells and remains resident with minimal cellular internalization for ~ 1.5 h. The successful plasma membrane-associated colabeling of the cells with the FsPFc10-FluoVolt™ donor-acceptor pair is confirmed by dual-channel confocal imaging. Importantly, cells labeled with FsPFc10 show excellent cellular viability with no adverse effect on cell membrane depolarization. During depolarization of membrane potential, HEK 293T/17 cells labeled with the donor-acceptor FRET pair exhibit a greater fluorescence response in FluoVolt™ emission relative to when FluoVolt™ is used as the sole imaging probe. These results demonstrate the conjugated polyelectrolyte to be a new class of membrane labeling fluorophore for use in voltage sensing schemes.

Enhanced Electron Transfer Mediated by Conjugated Polyelectrolyte and Its Application to Washing-Free DNA Detection.

Direct electron transfer between a redox label and an electrode requires a short working distance (<1-2 nm), and in general an affinity biosensor based on direct electron transfer requires a finely smoothed Au electrode to support efficient target binding. Here we report that direct electron transfer over a longer working distance is possible between (i) an anionic π-conjugated polyelectrolyte (CPE) label having many redox-active sites and (ii) a readily prepared, thin polymeric monolayer-modified indium-tin oxide electrode. In addition, the long CPE label (∼18 nm for 10 kDa) can approach the electrode within the working distance after sandwich-type target-specific binding, and fast CPE-mediated oxidation of ammonia borane along the entire CPE backbone affords high signal amplification.

An Ionic 1,4-Bis(styryl)benzene-Based Fluorescent Probe for Mercury(II) Detection in Water via Deprotection of the Thioacetal Group.

Highly sensitive and selective mercury detection in aqueous media is urgently needed because mercury poisoning usually results from exposure to water-soluble forms of mercury by inhalation and/or ingesting. An ionic conjugated oligoelectrolye (M1Q) based on 1,4-bis(styryl)benzene was synthesized as a fluorescent mercury(II) probe. The thioacetal moiety and quaternized ammonium group were incorporated for Hg2+ recognition and water solubility. A neutral Hg2+ probe (M1) was also prepared based on the same molecular backbone, and their sensor characteristics were investigated in a mixture of acetonitrile/water and in water. In the presence of Hg2+, the thioacetal group was converted to aldehyde functionality, and the resulting photoluminescence intensity decreased. In water, M1Q successfully demonstrated highly sensitive detection, showing a binding toward Hg2+ that was ~15 times stronger and a signal on/off ratio twice as high, compared to M1 in acetonitrile/water. The thioacetal deprotection by Hg2+ ions was substantially facilitated in water without an organic cosolvent. The limit of detection was measured to be 7 nM with a detection range of 10-180 nM in 100% aqueous medium.

Ratiometric fluorescent ion detection in water with high sensitivity via aggregation-mediated fluorescence resonance energy transfer using a conjugated polyelectrolyte as an optical platform.

A cationic conjugated polyelectrolyte was designed and synthesized based on poly(fluorene-co-phenylene) containing 5 mol% benzothiadiazole (BT) as a low energy trap and 15-crown-5 as a recognizing group for potassium ions. A potassium ion can form a sandwich-type 2:1 Lewis acid-based complex with 15-crown-5, to cause the intermolecular aggregation of polymers. This facilitates inter-chain fluorescence resonance energy transfer (FRET) to a low-energy BT segment, resulting in fluorescent signal amplification, even at dilute analyte concentrations. Highly sensitive and selective detection of K(+) ions was demonstrated in water. The linear response of ratiometric fluorescent signal as a function of [K(+) ] allows K(+) quantification in a range of nanomolar concentrations with a detection limit of ≈0.7 × 10(-9) M.