Comparative analysis of the absorption and fluorescence spectra and fluorescence excitation spectra of thioflavin T (ThT) in various solvents and in the composition of amyloid fibrils has shown that ThT, when excited in the region of the long-wavelength absorption band, fluoresces in the spectral region with a maximum at 478-484 nm. The appearance in aqueous and alcohol solutions of a fluorescence band with a maximum near 440 nm has been attributed to the presence in the composition of the ThT preparations of an impurity with an absorption band in the 340-350-nm range. The literature data showing that in glycerol ThT has a wide fluorescence spectrum with two maxima are due to the artifact connected with the use of a high concentration of the dye. It has been suggested that the cause of the low quantum yield of ThT aqueous and alcohol solutions is the breakage of the system of conjugated bonds due to the reorientation of the benzothiozole and benzaminic rings of ThT in the excited state with respect to one another. The main factor determining the high quantum yield of fluorescence of ThT incorporated in fibrils is the steric restriction of the rotation of the rings about one another under these conditions. The suggestions made have been verified by the quantumchemical calculation of the ThT molecule geometry in the ground and excited states.
The spectral-luminescent properties of symmetrical indotricarbocyanine dyes with the same cation and different anions (Br − , BF 4 − ) in cancerous HeLa cells, an isotonic solution of NaCl, and organic solvents have been investigated. It is shown that when these dyes are present in cells in nontoxic concentrations, they are in a monomeric state, they are localized in the region with a low dielectric permeability, bonded to biological macromolecules, and do not make contact with an aqueous medium. It has been established for the first time that the molecules of the polymeric dyes are present in cells predominantly in the form of contact ion pairs, and in a dye with the Br − anion the presence of free cations or solvately separated ionic pairs is revealed. It is shown that the shift of the ion equilibrium toward contact ion pairs for the dyes in the cells (as compared to the low-polarity organic solvents) can be due to the fairly high concentration of salts in the biosystem.
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