Styryl dyes have been among the most widely used probes for mapping membrane potential changes in excitable cells. However, their utility has been somewhat limited because their excitation wavelengths have been restricted to the 450–550 nm range. Longer wavelength probes can minimize interference from endogenous chromophores and, because of decreased light scattering, improve recording from deep within tissue. In this paper we report on our efforts to develop new potentiometric styryl dyes that have excitation wavelengths ranging above 700 nm and emission spectra out to 900 nm. We have prepared and characterized dyes based on 47 variants of the styryl chromophores. Voltage-dependent spectral changes have been recorded for these dyes in a model lipid bilayer and from lobster nerves. The voltage sensitivities of the fluorescence of many of these new potentiometric indicators are as good as those of the widely used ANEP series of probes. In addition, because some of the dyes are often poorly water soluble, we have developed cyclodextrin complexes of the dyes to serve as efficient delivery vehicles. These dyes promise to enable new experimental paradigms for in vivo imaging of membrane potential.
The effect of solvent vapor annealing on the fluorescence properties and morphology of titanyl phthalocyanine/ perylene phenethylimide thin-film molecular semiconductor bilayers (TiOPc/PPEI) is investigated. A combination of atomic force microscopy (AFM) and near-field scanning optical microscopy (NSOM) is used in conjunction with bulk absorption and fluorescence measurements to correlate the morphological and photophysical properties of these bilayer systems. AFM data show that treatment of the vacuum-deposited amorphous PPEI and TiOPc/PPEI films results in the crystalline transformation of these materials and severely alters the contact between the TiOPc and PPEI layers. AFM data show extended solvent vapor annealing produces void spaces in the TiOPc coverage on the order of several hundred nanometers. Steady-state fluorescence intensity and fluorescence lifetime measurements are used as a measure of charge-transfer quenching efficiencies. Very efficient charge-transfer quenching is observed when amorphous layers of TiOPc are deposited onto PPEI resulting from uniform contact between the layers. Extended annealing results in decreased charge-transfer quenching efficiencies as a result of widely dispersed, localized interfacial contact points.