Increased intensities of YOYO-1-labeled DNA oligomers near silver particles

DNA detection is usually performed using fluorescence probes. Using a DNA oligomer stained with the widely used dye 1,1'-[1,3-propanediylbis[(dimethylimino)-3,1-propanediyl]]bis[4-[(3-methyl-2(3H)-benzoxazolylidene)methyl]]-quinolinum tetraiodide (YOYO-1), we show that a substrate containing silver particles can lead to a greater than 10-fold increase in the fluorescence intensity. Proximity to silver particles also increases the photostability of YOYO-1-DNA. These results suggest that substrates or gels containing silver particles may be used for increased sensitivity in DNA detection.     

PICOSECOND RESOLUTION OF INDOLE ANISOTROPY DECAYS AND SPECTRAL RELAXATION BY 2 GHz FREQUENCY-DOMAIN FLUOROMETRY

Abstract— We used frequency-domain fluorescence spectroscopy to measure rotational diffusion and time-resolved emission spectra of indole in methanol on the picosecond timescale. The indole emission was quenched by acrylamide to allow measurements to the instrumental limit of 2 GHz and to eliminate emission from the longer-lived indole molecules, which can no longer provide information on the picosecond (ps) processes. The resolution was adequate to measure rotational correlation times as short as 8 ps at 80†C, and spectral relaxation times as short as 16 ps at 5†C.     

Evaluation of aging characteristics of bituminous roof coverings by thermo-mechanical analysis (TMA)

Four bituminous roof coverings have been subjected to various aging procedures including heat, UV irradiation and temperature/rain cycling. A specially modified TMA technique has been used for evaluation, depending on measurements of softening temperature. A comparison with mechanical test data has also been made. Some conclusions have been drawn with respect to influence of heat and UV radiation on the rate of degradation.     

Distributions of fluorescence decay times for synthetic melittin in water-methanol mixtures and complexed with calmodulin,troponin C,and phospholipids

Frequency-domain measurements of the intensity decays of melittin were used to recover the distribution of decay times displayed by its single tryptophan residue. Melittin was examined in the monomeric random coil state (water), in the monomeric -helical state (water-methanol), in the tetrameric state, and with 6M guanidine hydrochloride. In the presence of denaturant, where melittin is expected to be devoid of secondary structure, we observed a narrow distribution of lifetimes, similar to a double-exponential decay. In water the intensity decay of melittin was found to be described better by the distribution of decay times, which became progressively wider as the amount of -helix was increased by the methanol cosolvent or upon formation of the -helical tetrameric state. We also examined the intensity decays of melittin when complexed with calmodulin, troponin C, or lipid vesicles of 1-palmitoyl-2-oleyl-l--phosphatidylcholine (POPC). The lifetime distributions of the complexes with lipid were comparable to those observed in methanol-water, suggesting a similarity of the structure and/or dynamics of the environment surrounding the tryptophan residue. A broad lifetime distribution was observed for the melittin-calmodulin complex, suggesting a rigid structure and/or heterogeneity in the form of the complex. The lifetime distribution of the melittin-troponin C complex was more narrow, suggesting a more uniform structure, at least in the region surrounding the tryptophan residue. These results demonstrate that the lifetime distributions of a single tryptophan protein can be a sensitive indicator of the conformational heterogeneity and dynamics of proteins.     

Multiphoton Ligand-Enhanced Excitation of Lanthanides

We describe multiphoton excitation of the lanthanides europium (Eu³⁺) and terbium (Tb³⁺) when these ions are complexed with nucleic acids, proteins, and fluorescent chelators. In all cases excitation occurs by multiphoton absorption of the sensitizers. For the nucleotide GDP and an oligonucleotide with several guanines, the sensitized emission of Tb³⁺ excited at 776 nm indicated a three-photon process. For Tb³⁺ bound to the wild-type troponin C and a single tryptophan mutant (26W), excitation at 794 nm was also close to a three-photon process. For lanthanide chelators containing various sensitizers, we observed three-photon excitation in the case of methyl anthranilate, a mixuture of two- and three-photon excitation for carbostyril 124, and a two-photon process with a coumarin derivative. In the case of coumarin-sensitized emission of Eu³⁺ varied from a two- to a three-photon process at wavelengths ranging from 780 to 880 nm. The sensitized luminescence also shows significantly higher photostability compared to the fluorescence from the organic fluorophores alone. These results suggest the use of multiphoton-induced sensitized lanthanide fluorescence in biochemistry and cellular imaging.On leave from the Institute of Experimental Physics     

Förster energy transfer from nonexponentially decaying donors

Necessary modifications to the expression for the F?rster energy transfer rate are discussed when fluorescence decay of the donor in the absence of acceptor is nonexponential. Discrete and continuous models of the nonexponentiality are taken into account. No general solution of the problem is found. It is, however, suggested that in many of the biochemical problems the most appropriate modification of the transfer rate can be that which is based on the assumption of the same constant value of the radiative decay rate for all donor molecules. The effect of the assumed form of the F?rster energy transfer rate on the recovered values of the distance distribution and dynamics parameters of some exemplary bichromophoric systems is examined.     

Exploring fundamental differences between red- and blue-shifted intramolecular hydrogen bonds using FAMSEC,FALDI, IQA and QTAIM

We have discovered, using developed by us recently FALDI and FAMSEC computational techniques, fundamentally distinct mechanisms of intramolecular red- and blue-shifted H-bond formation that occurred in different conformers of the same molecule (amino-acid β-alanine) involving the same heteroatoms (O–H???N and N–H???O). Quantitative topological, geometric and energetic data of both H-bonds obtained with well-known QTAIM and IQA methodologies agree with what is known regarding H-bonding in general. However, the FALDI charge and decomposition scheme for calculating in real space 3D conformational deformation densities provided clear evidence that the process of electron density redistribution taking place on the formation of the stronger red-shifted H-bond is fundamentally distinct from the weaker blue-shifted H-bond. Contributions made by atoms of the X–H???Y–Z fragment (IUPAC notation) as well as distinct atoms on the H-bond formation were fully explored. The FAMSEC energy decomposition approach showed that the atoms involved in formation of the red-shifted H-bond interact in a fundamentally different fashion, both locally and with the remainder of the molecule, as compared with those of the blue-shifted H-bond. Excellent correlations of trends obtained with QTAIM, IQA, FAMSEC and FALDI techniques were obtained. Commentary regarding IUPAC recommended definition of an H-bond and validity of observed AILs (or bond paths) of the two H-bond kinds is also discussed.