Directionality of light absorption and emission in representative fluorescent proteins

Fluorescent molecules are like antennas: The rate at which they absorb light depends on their orientation with respect to the incoming light wave, and the apparent intensity of their emission depends on their orientation with respect to the observer. However, the directions along which the most important fluorescent molecules in biology, fluorescent proteins (FPs), absorb and emit light are generally not known. Our optical and X-ray investigations of FP crystals have now allowed us to determine the molecular orientations of the excitation and emission transition dipole moments in the FPs mTurquoise2, eGFP, and mCherry, and the photoconvertible FP mEos4b. Our results will allow using FP directionality in studies of molecular and biological processes, but also in development of novel bioengineering and bioelectronics applications.

Airglow Derived Measurements of Q-Branch Transition Probabilities for Several Hydroxyl Meinel Bands

Spectroscopic measurements of the hydroxyl (OH) airglow emissions are often used to infer neutral temperatures near the mesopause. Correct Einstein coefficients for the various transitions in the OH airglow are needed to calculate accurate temperatures. However, studies from some studys showed experimentally and theoretically that the most commonly used Einstein spontaneous emission transition probabilities for the Q-branch of the OH Meinel (6,2) transition are overestimated. Extending their work to several Δv = 2 and 3 transitions from v′ = 3 to 9, we have determined Einstein coefficients for the first four Q-branch rotational lines. These have been derived from high resolution, high signal to noise spectroscopic observations of the OH airglow in the night sky from the Nordic Optical Telescope. The Q-branch Einstein coefficients calculated from these spectra show that values currently tabulated in the HITRAN database overestimate many of the Q-branch transition probabilities. The implications for atmospheric temperatures derived from OH Q-branch measurements are discussed.

Fluorescence anisotropy imaging of a polydiacetylene photopolymer film

UV-illumination of phase-separated surfactant films prepared from mixtures of photopolymerizable 10,12-pentacosadiynoic acid and perfluorotetradecanoic acid results in the formation of fluorescent polydiacetylene fibers and aggregates. In this work, the orientation of polymer strands that comprise the resulting photopolymer structures has been probed using fluorescence anisotropy imaging in combination with defocused single-molecule fluorescence imaging. Imaging experiments indicate the presence of significant fiber-to-fiber heterogeneity, as well as anisotropy within each fiber (or aggregate), with both of these properties changing as a function of film preparation conditions. This anisotropy can be attributed to various alignments of the constituent polymer strands that comprise the larger fibers and aggregates. Intriguingly, when using defocused imaging, fiber images consisted of a series of discrete “doughnut” fluorescence emission patterns, which exhibited intermittent on–off blinking behavior; both of these properties are characteristic of individual emission transition dipoles (single molecules). Further, all of the individual emission transition dipoles had a uniform orientation with respect to the axis of the fiber, indicating a common orientation of discrete emitters in the larger polymer fiber. The implications of these results for future studies of the electronic properties of conjugated polymers in larger macroscopic systems are noted.

NIR fluorescent organic nanoparticles for photoinduced nitric oxide delivery with self monitoring and real time reporting abilities

Nitric oxide photodonor (NOD) conjugated perylene tetracarboxylate ester (TPT) based fluorescent organic TPT(NOD)4 nanoparticles (NPs) with aggregation induced NIR emission have shown photoinduced nitric oxide delivery along with a red to green emission transition.