Ultrafast Radical Photogeneration Pathways in Eumelanin†

Eumelanin is a ubiquitous biological pigment that rapidly and efficiently deactivates excited states created by UV or visible radiation. Paradoxically, photoirradiation of eumelanin also generates radicals and harmful reactive oxygen species, but the relationship between these pathways and excited-state deactivation is uncertain. Here, greatly expanding the excitation tuning range (225–620 nm) and probing window (400–1500 nm) in femtosecond transient absorption spectroscopy of the synthetic eumelanin, DOPA melanin, enables the detection of photogenerated radials with ultrafast time resolution for the first time. Despite its heterogeneous nature, the transient absorption signals can be modeled by two spectral components assigned to solvated electrons and photogenerated radicals. Radical absorbance measured several nanoseconds after excitation increases exponentially with increasing photon energy, matching the trend in radical yields measured in electron paramagnetic resonance spectroscopy experiments. Spectral modeling of the transient signals reveals two radical generation mechanisms: (1) photoionization by UV light; and (2) photoinduced charge transfer among eumelanin chromophores by UVA and visible wavelengths capable of reaching the pigment in skin. Concurrent ultrafast relaxation and radical generation underlie the ability of eumelanin to be both photoprotective and photodamaging, and the branching between these pathways likely depends on the wavelength of the absorbed light.     

Two-color visible/vacuum ultraviolet photoelectron imaging dynamics of Br2

An experimental two-color photoionization dynamics study of laser-excited Br2 molecules is presented, combining pulsed visible laser excitation and tunable vacuum ultraviolet (VUV) synchrotron radiation with photoelectron imaging. The X 1Sigmag + -B 3Pi0+u transition in Br2 is excited at 527 nm corresponding predominantly to excitation of the v' = 28 vibrational level in the B 3Pi0+u state. Tunable VUV undulator radiation in the energy range of 8.40-10.15 eV is subsequently used to ionize the excited molecules to the X 2Pi32,12 state of the ion, and the ionic ground state is probed by photoelectron imaging. Similar experiments are performed using single-photon synchrotron ionization in the photon energy range of 10.75-12.50 eV without any laser excitation. Photoelectron kinetic energy distributions are extracted from the photoelectron images. In the case of two-color photoionization using resonant excitation of the intermediate B 3Pi0+u state, a broad distribution of photoelectron kinetic energies is observed, and in some cases even a bimodal distribution, which depends on the VUV photon energy. In contrast, for single-photon ionization, a single nearly Gaussian-shaped distribution is observed, which shifts to higher energy with photon energy. Simulated spectra based on Franck-Condon factors for the transitions Br2(X 1Sigmag+, v" = 0)-Br2 +(X 2Pi12,32, v+) and Br2(B 3Pi0+u, v' = 28)-Br2 +(X 2Pi12,32, v+) are generated. Comparison of these calculated spectra with the measured images suggests that the differences in the kinetic energy distributions for the two ionization processes reflect the different extensions of the vibrational wave functions in the v" = 0 electronic ground state (X 1Sigmag+) versus the electronically and vibrationally excited state (B 3Pi0+u, v' = 28).     

Femtosecond Two-photon Excited Fluorescence of Melanin*

Fluorescence of synthetic melanin in dimethyl sulfoxide has been excited by two-photon absorption at 800 nm, using 120 fs pulses with photon flux densities > or = 10(27) cm-2 s-1. The shortest main component of the three-exponential decay of fluorescence is 200 +/- 2 ps. The overall spectral shape is red-shifted with respect to the 400 nm excited fluorescence. Two-photon excited melanin fluorescence also has been measured from excised samples of healthy human skin tissue. Because of the selectivity of melanin excitation via resonant two-photon absorption, it is hypothesized that fluorescence excited in this way may yield information on malignant transformation.     

Two-photon and multi-photon excitation of naphthalene in the vapor phase,studied by excitation and fluorescence spectra

Naphthalene vapor is irradiated by μsec dye laser pulses of 150 kW peak power and a spectral bandwidth of 0.3 nm. A two-photon excitation spectrum is detected by monitoring the near UV fluorescence as a function of laser wavelength which is tuned between 570 and 610 nm. The fluorescence obtained by irradiation into the strongest band of the two-photon spectrum could be spectroscopically resolved using a bandwidth of 80 cm^?1. The spectrum exhibits vibrational structure which lies on a strong non-resolved background. From information in both spectra it can be definitely concluded that vibronic levels of B_3u × b_3u species in the lowest singlet state are predominantly excited in a two-photon process. The non-resolved background in the fluorescence spectrum is attributed to subsequent excitation of the two-photon state by a third photon. Further stepwise excitation in the strong radiation field of the laser is also taken into account.     

Light-driven decarboxylation of wild-type green fluorescent protein

The response of wild-type GFP to UV and visible light was investigated using steady state absorption, fluorescence, and Raman spectroscopies. As reported previously [van Thor, Nat. Struct. Biol. 2002, 9, 37-41], irradiation of GFP results in decarboxylation of E222. Here it is reported that the rate of the light-driven decarboxylation reaction strongly depends on the excitation wavelength, decreasing in the order 254 nm > 280 nm > 476 nm. The relative efficiencies of decarboxylation are explained in terms of the Kolbe-type mechanism in which the excited state of the chromophore acts as an oxidant by accepting an electron from E222. Specifically, it is proposed that 254 nm excitation populates the S2 (or higher) excited state of the chromophore, whereas 404 and 476 nm excitation populate the S1 excited state of neutral and anionic forms, respectively, and that the relative oxidizing power of the three excited states controls the rate of the decarboxylation reaction. In addition, the role of W57 in the photophysics of GFP has been probed by mutating this residue to phenylalanine. These studies reveal that while W57 does not affect decarboxylation, this residue is involved in resonance energy transfer with the chromophore, thereby partially explaining the green fluorescence observed upon UV irradiation of wild-type GFP. Finally, comparison of Raman spectra obtained from nonilluminated and decarboxylated forms of wild-type GFP has provided further vibrational band assignments for neutral and anionic forms of the chromophore within the protein. In addition, these spectra provide valuable insight into the specific interactions between the protein and the chromophore that control the optical properties of wild-type GFP.     

Multi‐Photon Excitation in Photoredox Catalysis: Concepts,Applications, Methods

The energy of visible photons and the accessible redox potentials of common photocatalysts set thermodynamic limits to photochemical reactions that can be driven by traditional visible‐light irradiation. UV excitation can be damaging and induce side reactions, hence visible or even near‐IR light is usually preferable. Thus, photochemistry currently faces two divergent challenges, namely the desire to perform ever more thermodynamically demanding reactions with increasingly lower photon energies. The pooling of two low‐energy photons can address both challenges simultaneously, and whilst multi‐photon spectroscopy is well established, synthetic photoredox chemistry has only recently started to exploit multi‐photon processes on the preparative scale. Herein, we have a critical look at currently developed reactions and mechanistic concepts, discuss pertinent experimental methods, and provide an outlook into possible future developments of this rapidly emerging area.     

The photodissociation of NO(2) by visible and ultraviolet light

We present velocity map images of the NO, O((3)P(J)) and O((1)S(0)) photofragments from NO(2) excited in the range 7.6 to 9.0 eV. The molecule was initially pumped with a visible photon between 2.82-2.95 eV (440-420 nm), below the first dissociation threshold. A second ultraviolet laser with photon energies between 4.77 and 6.05 eV (260-205 nm) was used to pump high-lying excited states of neutral NO(2) and/or probe neutral photoproducts. Analysis of the kinetic energy release spectra revealed that the NO photofragments were predominantly formed in their ground electronic state with little kinetic energy. The O((3)P(J)) and O((1)S(0)) kinetic energy distributions were also dominated by kinetically 'cold' fragments. We discuss the possible excitation schemes and conclude that the unstable photoexcited states probed in the experiment were Rydberg states coupled to dissociative valence states. We compare our results with recent time-resolved studies using similar excitation and probe photon energies.     

Photoprotection of Mammalian Acid-Soluble Collagen by Cuttlefish Sepia Melanin In Vitro

Several important clinical conditions can result in close association between the pigment melanin and dermal collagen. Because melanin and its precursors can be chemically reactive in ground and excited states, it is important to know whether the resulting melanin-collagen interaction results in photoprotection or photoaggression. Acidic and neutral air-saturated collagen suspensions (0.033%) were irradiated with0–2.6 times 104 J/m2 UVC or with0–83 times 104 J/m2 solar-simulating UV radiation (SSR). Photochemical destruction of a photolabile collagen fluorophore (δ_em 360 nm) and collagen chain degradation were monitored as functions of irradiation time in the presence and absence of added (0–100μg) sepia eumelanin. Melanin retarded collagen photodamage but did not qualitatively alter the fluorescence fading kinetics. Both H202 and 02 can be produced by UV irradiation of eumelanin. Added H202 and K02 destroyed collagen fluorescence and caused 50% chain degradation at ca10–20-fold molar excess. Previous studies have demonstrated that eumelanins efficiently scavenge 02 . We demonstrated that eumelanin also efficiently scavenges H202 as evidenced by its ability to (a) compete with scopoletin for peroxide uptake and (b) directly take up H202 through a dialysis bag. The latter observation suggests that peroxide scavenging could occur in vivo by melanin sequestered in melanophages. Thus, neither UV-generated 02 nor H202 are likely to be present in concentrations high enough to cause measurable collagen damage. Absorption and/or scattering of excitation radiation away from the target chromophore appears to be the primary photoprotection mechanism, although scavenging of active 02 intermediates may play an important, if subtle role.     

HREELS to identify electronic structures of organic thin films

The electronic structure of alpha-oligothiophene (alphanT) thin films has been investigated for increasing chain lengths of n= 4-8 thiophene units with high resolution electron energy loss spectroscopy (HREELS) in the specular reflection geometry at a primary energy of 15 eV. The great advantage of this technique in contrast to UV/VIS absorption spectroscopy results from the fact, that the impact scattering mechanism of HREELS makes it possible to also detect optically forbidden electronic transitions. On the other hand, the electrons used as probes in HREELS have a wavelength which is two orders of magnitudes smaller if compared to those of photons used in UV/VIS absorption spectroscopy. Therefore individual molecules are excited by HREELS independent from each other and hence the excitation of collective excitons is not possible. As a result, information about the orientation of the molecules cannot be achieved with HREELS, which, however, is possible in polarization-dependent UV/VIS spectroscopy.     

Two‐photon excitation in chip electrophoresis enabling label‐free fluorescence detection in non‐UV transparent full‐body polymer chips

One of the most commonly employed detection methods in microfluidic research is fluorescence detection, due to its ease of integration and excellent sensitivity. Many analytes though do not show luminescence when excited in the visible light spectrum, require suitable dyes. Deep‐ultraviolet (UV) excitation (<300 nm) allows label‐free detection of a broader range of analytes but also mandates the use of expensive fused silica glass, which is transparent to UV light. Herein, we report the first application of label‐free deep UV fluorescence detection in non‐UV transparent full‐body polymer microfluidic devices. This was achieved by means of two‐photon excitation in the visible range (λ_ex = 532 nm). Issues associated with the low optical transmittance of plastics in the UV range were successfully circumvented in this way. The technique was investigated by application to microchip electrophoresis of small aromatic compounds. Various polymers, such as poly(methyl methacrylate), cyclic olefin polymer, and copolymer as well as poly(dimethylsiloxane) were investigated and compared with respect to achievable LOD and ruggedness against photodamage. To demonstrate the applicability of the technique, the method was also applied to the determination of serotonin and tryptamine in fruit samples.     

The role of melanin as protector against free radicals in skin and its role as free radical indicator in hair

Throughout the body, melanin is a homogenous biological polymer containing a population of intrinsic, semiquinone-like radicals. Additional extrinsic free radicals are reversibly photo-generated by UV and visible light. Melanin photochemistry, particularly the formation and decay of extrinsic radicals, has been the subject of numerous electron spin resonance (ESR) spectroscopy studies. Several melanin monomers exist, and the predominant monomer in a melanin polymer depends on its location within an organism. In skin and hair, melanin differs in content of eumelanin or pheomelanin. Its bioradical character and its susceptibility to UV irradiation makes melanin an excellent indicator for UV-related processes in both skin and hair. The existence of melanin in skin is strongly correlated with the prevention against free radicals/ROS generated by UV radiation. Especially in the skin melanin (mainly eumelanin) ensures the only natural UV protection by eliminating the generated free radicals/ROS. Melanin in hair can be used as a free radical detector for evaluating the efficacy of hair care products. The aim of this study was to investigate the suitability of melanin as protector of skin against UV generated free radicals and as free radical indicator in hair.     

INDUCTION OF LAMBDA PROPHAGE NEAR THE SITE OF FOCUSED UV LASER RADIATION

DNA damage from photon scatter or beam spread during UV excimer laser irradiation was investigated using the induction of bacteriophage lambda in E. coli BR339. Prophage induction in these cells leads to the production of beta-galactosidase which can be detected colorimetrically by the application of appropriate substrates. An agar surface overlayed with BR339 cells was placed at various distances from the focal point of a converging lens and exposed to either 193 or 248 nm laser radiation. Energy densities ranging from approximately 5 mJ/cm2 to 30 J/cm2 were used. Ablation with 193 nm laser radiation produced an 800 microns wide clear 'trench' surrounded by a 500 microns zone of cells in which lambda had been induced. Following ablation with 248 nm laser radiation, the zone of induction was several millimeters wide. Exposures to 193 nm radiation at 170 mJ/cm2/pulse produced visible ablation of the agar surface at 1.7 J/cm2. Lambda induction was observed surrounding cleared ablation areas. The presence of induction in this system suggests that both 248 and 193 nm excimer laser radiation delivered at high energy densities has sufficient spread or scatter to damage DNA in cells surrounding areas of ablation.     

Action Spectra for UVB Impacts on Blepharisma japonicum Motility and Photobehavior

Abstract— The ciliate Blepharisma japonicum was exposed to artificial polychromatic and monochromatic UV radiation to evaluate the relative roles of UVB (280–315 nm UV radiation) and UVA (315–400 nm UV radiation) in altering its motility and photobehavior and to determine absolute weighting coefficients for these effects in the UVB range. Under polychromatic UV irradiation B. japonicum cells showed a severe reduction of cell speed and of the capability to respond to light stimuli. At low doses, however, UV caused a significant increase in the average velocity of a cell population. The UVB exclusion experiments indicated that UVA does not significantly alter motility and photoresponsiveness. The increase and the subsequent decrease in cell velocity was observed also under monochromatic irradiation at 281, 290 and 300 nm, whereas at 310 nm cells swim faster up to the highest photon flux density used. The cell capability of reacting to photic stimuli, conversely, steadily declined with increasing photon flux density at all the tested UVB wavelengths. The action spectra for the alteration of cell velocity and the impairment of photoresponsiveness show that the lower the irradiation wavelength, the more remarkable are the UVB effects and suggest different targets for the increase and the decrease in cell velocity.     

SrB4O7:Pr^3+中Pr^3+的发光性质

本研究报道Pr^3+在SrB4O7中的发光性质,在SrB4O7中Pr^3+离子的4f5d能态高于1^S0能级,因此,在207nmUV光激发下,Pr^3+能够把所吸收的一个高能量的UV光子转换为两个可见光子的发射(光子倍增);在此氧化物基质中的光子倍增主要是由于阳离子处于弱的晶体场格位之中;由于与稀土离子弱联结相关的声子振动频率低(hωmax～1200cm^-1),因此还能观察到从3^P0能级向低能级的跃迁。第一个光子的发射由1^S0→1^G4(313nm),1^S0→1^D2(338nm)和1^S0→1^I6(405nm)的辐射跃迁组成;第二个光子的发射由3^P0和1^D2能级向低能级的辐射跃迁组成[3^P0→(3^HJ,3^FJ)和1^D2→(3^H4,3^H5)]。     

White‐Light Emission from Dual‐Way Photon Energy Conversion in a Dye‐Encapsulated Metal–Organic Framework

The design of white‐light phosphors is attractive in solid‐state lighting (SSL) and related fields. A new strategy in obtaining white light emission (WLE) from dual‐way photon energy conversion in a series of dye@MOF ( LIFM‐WZ‐6 ) systems is presented. Besides the traditional UV‐excited one‐photon absorption (OPA) pathway, white‐light modulation can also be gained from the combination of NIR‐excited green and red emissions of MOF backbone and encapsulated dyes via two‐photon absorption (TPA) pathway. As a result, down‐conversion OPA white light was obtained for RhB⁺@LIFM‐WZ‐6 (0.1 wt %), BR‐2⁺@LIFM‐WZ‐6 (2 wt %), and APFG⁺@LIFM‐WZ‐6 (0.1 wt %) samples under 365 nm excitation. RhB⁺@LIFM‐WZ‐6 (0.05 wt %), BR‐2⁺@LIFM‐WZ‐6 (1 wt %) and APFG⁺@LIFM‐WZ‐6 (0.05 wt %) exhibit up‐conversion TPA white light under the excitation of 800, 790, and 730 nm, respectively. This new WLE generation strategy combines different photon energy conversion mechanisms together.     

Prolonged Hot Electron Dynamics in Plasmonic‐Metal/Semiconductor Heterostructures with Implications for Solar Photocatalysis

Ideal solar‐to‐fuel photocatalysts must effectively harvest sunlight to generate significant quantities of long‐lived charge carriers necessary for chemical reactions. Here we demonstrate the merits of augmenting traditional photoelectrochemical cells with plasmonic nanoparticles to satisfy these daunting photocatalytic requirements. Electrochemical techniques were employed to elucidate the mechanics of plasmon‐mediated electron transfer within Au/TiO₂ heterostructures under visible‐light (λ>515 nm) irradiation in solution. Significantly, we discovered that these transferred electrons displayed excited‐state lifetimes two orders of magnitude longer than those of electrons photogenerated directly within TiO₂ via UV excitation. These long‐lived electrons further enable visible‐light‐driven H₂ evolution from water, heralding a new photocatalytic paradigm for solar energy conversion.     

TRIPLET YIELD OF MEROCYANINE 540 IN WATER IS WAVELENGTH DEPENDENT

Monomers and aggregates of Merocyanine 540 (MC540) in water are able to photoisomerize. The shape of the photoisomer absorption spectrum is very similar to that of the ground state. Triplet state of MC540 in water has been produced by energy transfer from triplet anthracene and displays a broad absorption spectrum between 600 and 700 nm. The triplet state may also be produced by direct excitation of MC540 with UV light. However, when the dye is excited by visible light, no triplet state absorbance in the red could be detected so that the triplet yield of MC540 in water seems to be excitation wavelength dependent.     

Radical Protection by Differently Composed Creams in the UV/VIS and IR Spectral Ranges

Modern sunscreens are well suited to provide sufficient protection in the UV range because the filter substances absorb or scatter UV radiation. Although up to 50% of radicals are formed in the visible and infrared spectral range during solar radiation protection strategies are not provided in this range. Previous investigations of commercially available products have shown that in addition to physical filters, antioxidants (AO) are necessary to provide protective effects in the infrared range by neutralizing already formed radicals. In this study, the efficacy of filter substances and AO to reduce radical formation in both spectral ranges was investigated after UV/VIS or IR irradiation. Optical properties and radical protection were determined for the investigated creams. It was found that organic UV filters lower radical formation in the UV/VIS range to 35% compared to untreated skin, independent of the presence of AO. Further reduction to 14% was reached by addition of 2% physical filters, whereas physical filters alone were ineffective in the UV/VIS range due to the low concentration. In contrast, this filter type reduced radical formation in the IR range significantly to 65%; similar effects were aroused after application of AO. Sunscreens which contain organic UV filters, physical filters and AO ensure protection in the complete solar spectrum.     

Evidence from action and fluorescence spectra that UV-induced violet-blue-green fluorescence enhances leaf photosynthesis

We assessed the contribution of UV-induced violet-blue-green leaf fluorescence to photosynthesis in Poa annua, Sorghum halepense and Nerium oleander by measuring UV-induced fluorescence spectra (280-380 nm excitation, 400-550 nm emission) from leaf surfaces and determining the monochromatic UV action spectra for leaf photosynthetic O2-evolution. Peak fluorescence emission wavelengths from leaf surfaces ranged from violet (408 nm) to blue (448 nm), while excitation peaks for these maxima ranged from 333 to 344 nm. Action spectra were developed by supplementing monochromatic radiation from 280 to 440 nm, in 20 nm increments, to a visible nonsaturating background of 500 mumol m-2 s-1 photosynthetically active radiation and measuring photosynthetic O2-evolution rates. Photosynthetic rates tended to be higher with the 340 nm supplement than with higher or lower wavelength UV supplements. Comparing photosynthetic rates with the 340 nm supplement to those with the 400 nm supplement, the percentage enhancement in photosynthetic rates at 340 nm ranged from 7.8 to 9.8%. We suspect that 340 nm UV improves photosynthetic rates via fluorescence that provides violet-blue-green photons for photosynthetic energy conversion because (1) the peak excitation wavelength (340 nm) for violet-blue-green fluorescence from leaves was also the most effective UV wavelength at enhancing photosynthetic rates, and (2) the magnitude of photosynthetic enhancements attributable to supplemental 340 nm UV was well correlated (R2 = 0.90) with the apparent intensity of 340 nm UV-induced violet-blue-green fluorescence emission from leaves.     

Theoretical study toward understanding ultrafast internal conversion of excited 9H-adenine

The CASPT2/CASSCF method with the 6-311G basis set and an active space up to (14, 11) was used to explore the ultrafast internal conversion mechanism for excited 9H-adenine. Three minima, two transition states, and seven conical intersections were obtained to build up the two deactivation pathways for the internal conversion mechanism. Special efforts were made to explore the excited-state potential energy surfaces near the Franck-Condon region and determine the various barriers in the processes of deactivation. The barrier required from the 1pipi (1La) state to deactivate nonradiatively is found to be lower than that required from the 1pipi (1Lb) state. On 250 nm excitation, the 1pipi (1La) state is populated, and the transition from 1pipi (1La) to the lowest 1npi state involves very low barriers, which may account for the observed short (<50 fs) lifetime of the 1pipi excited state. The deactivation of the lowest 1npi state is required to overcome a barrier of 3.15 kcal/mol, which should be responsible for the 750 fs lifetime of the npi excited state. On 267 nm excitation, the vibrationally active 1pipi (1Lb) state is populated. Excitation at 277 nm prepares the 1pipi (1Lb) state without much excessive vibrational energy, which may be responsible for the observed >2 ps lifetime.