Influence of Surface Structure, Pigmentation and Particulate Matter on Plant Reflectance and Fluorescence.

Optical properties of plant leaves are relevant to evaluate their physiological state and stress effect. The main objective of this work was to study how variegation, pigment composition or reflective features modifies leaves' photophysical behavior. For this purpose, green leaves (Ficus benjamina), purple leaves (Tradescantia pallida), green leaves covered by white trichomes (Cineraria maritima) and variegated leaves (Codiaeum aucubifolium) were analyzed. Firstly, foliar surface morphology was evaluated by scanning electron microscopy. UV-vis and near-IR reflectance and transmittance spectra were obtained to calculate absorption (k) and scattering (s) coefficients. The theoretical approaches of Pile of Plates and Kubelka-Munk's theory resulted still valid for nonstandard leaves with differing surface conditions. However, frequently used spectral indices were not reliable for predicting water content, when leaves differed from conventional ones. The proportionality between the absorption factor and chromophore/pigment concentration was also lost for hairy leaves. A simplified model to describe these facts was presented here. Fluorescence spectra were recorded and corrected, due to light re-absorption. Water-optical parameter connection and pigment-optical parameter connection were thoroughly discussed. Leaf surface morphology and pigmentation have not only influenced the optical features of leaves but also played a role in the effect that particulate matter could cause on leaf photosynthesis.

Photophysics at Unusually High Dye Concentrations.

The study of the interaction of light with systems at high dye concentrations implies a great challenge because several factors, such as emission reabsorption, dye aggregation, and energy trapping, hinder rationalization and interpretation of the involved photophysical processes. Space constraints induce dye interaction even in the absence of ground state stabilization of dimers and oligomers. At distances on the order of 1 nm, statistical energy traps are usually observed. At longer distances, excited state energy transfer takes place. Most of these factors do not result in ground state spectroscopic changes. Rather, fluorescence phenomena such as inner filter effects, concentration-dependent Stokes' shifts, and changes in quantum yields and decay times are evidenced. Photophysical studies are commonly carried out at high dilution, to minimize dye-dye interactions and emission reabsorption, and in the absence of light scattering. Under these conditions, the physical description of the system becomes rather simple. Fluorescence and triplet quantum yields become molecular properties and can be easily related to ratios of rate constants. However, many systems containing dyes able to fulfill specific functions, whether man-made or biological, are far from being dilute and scattering-free. The photosynthetic apparatus is a paradigmatic example. It is clear that isolation of components allows gathering relevant information about complex systems. However, knowledge of the photophysical behavior in the unaltered environment is essential in most cases. Complexity generally increases when light scattering is present. Despite that, our experience shows that light scattering, when correctly handled, may even simplify the task of unraveling molecular parameters. We show that methods and models aiming at the determination and interpretation of fluorescence and triplet quantum yields as well as energy transfer efficiencies can be developed on the basis of simple light-scattering theories. Photophysical studies were extended to thin films and layer-by-layer assemblies. Procedures are presented for the evaluation of fluorescence reabsorption in concentrated fluid solutions up to the molar level, which are being applied to ionic liquids, in which the emitting species are the bulk ions. Fluorescence reabsorption models proved to be useful in the interpretation of the photophysics of living organisms such as plant leaves and fruits. These new tools allowed the assessment of chlorophyll fluorescence at the chloroplast, leaf and canopy levels, with implications in remote sensing and the development of nondestructive optical methods.

Photophysics of Xanthene Dyes at High Concentrations in Solid Environments: Charge Transfer Assisted Triplet Formation.

The photophysical behavior of two xanthene dyes, Eosin Y and Phloxine B, included in microcrystalline cellulose particles is studied in a wide concentration range, with emphasis on the effect of dye concentration on fluorescence and triplet quantum yields. Absolute fluorescence quantum yields in the solid-state were determined by means of diffuse reflectance and steady-state fluorescence measurements, whereas absolute triplet quantum yields were obtained by laser-induced optoacoustic spectroscopy and their dependence on dye concentration was confirmed by diffuse reflectance laser flash photolysis and time-resolved phosphorescence measurements. When both quantum yields are corrected for reabsorption and reemission of radiation, ΦF values decrease strongly on increasing dye concentration, while a less pronounced decay is observed for ΦT . Fluorescence concentration quenching is attributed to the formation of dye aggregates or virtual traps resulting from molecular crowding. Dimeric traps are however able to generate triplet states. A mechanism based on the intermediacy of charge-transfer states is proposed and discussed. Calculation of parameters for photoinduced electron transfer between dye molecules within the traps evidences the feasibility of the proposed mechanism. Results demonstrate that photoactive energy traps, capable of yielding dye triplet states, can be formed even in highly-concentrated systems with random dye distributions.

Rose bengal in poly(2-hydroxyethyl methacrylate) thin films: self-quenching by photoactive energy traps.

The effect of dye concentration on the fluorescence,ΦF, and singlet molecular oxygen,ΦΔ, quantum yields of rose bengal loaded poly(2-hydroxyethyl methacrylate) thin films (∼200 nm thick) was investigated, with the aim of understanding the effect of molecular interactions on the photophysical properties of dyes in crowded constrained environments. Films were characterized by absorption and fluorescence spectroscopy, singlet molecular oxygen (1O2) production was quantified using a chemical monitor, and the triplet decay was determined by laser flash-photolysis. For the monomeric dilute dye, ΦF = 0.05 ± 0.01 and ΦΔ = 0.76 ± 0.14. The effect of humidity and the photostability of the dye were also investigated. Spectral changes in absorption and fluorescence in excess of 0.05 M and concentration self-quenching after 0.01 M are interpreted in the context of a quenching radius model. Calculations of energy migration and trapping rates were performed assuming random distribution of the dye. Best fits of fluorescence quantum yields with concentration are obtained in the whole concentration range with a quenching radius r Q = 1.5 nm, in the order of molecular dimensions. Agreement is obtained only if dimeric traps are considered photoactive, with an observed fluorescence quantum yield ratio ΦF,trap/ΦF,monomer ≈ 0.35. Fluorescent traps are capable of yielding triplet states and 1O2. Results show that the excited state generation efficiency, calculated as the product between the absorption factor and the fluorescence quantum yield, is maximized at around 0.15 M, a very high concentration for random dye distributions. Relevant information for the design of photoactive dyed coatings is provided.

Tuning the concentration of dye loaded polymer films for maximum photosensitization efficiency: phloxine B in poly(2-hydroxyethyl methacrylate).

Fluorescence and singlet molecular oxygen ((1)O2) quantum yields for phloxine B loaded poly(2-hydroxyethyl methacrylate) thin films are determined at dye concentrations from 0.015 to 22 wt%. Fluorescence self-quenching and the fall off of the (1)O2 quantum yield observed above 0.1 wt% are attributed to very weakly interacting close-lying dye molecules acting as energy traps arising from molecular confinement. The maximum singlet oxygen generation efficiency (quantum yield × absorption factor) lies at concentrations around 2 wt%, where fluorescence self quenching amounts to more than 80%. Data are fitted quantitatively by using a quenching radius model involving energy migration and trapping with rQ = 1.2 nm. The present results constitute a proof of concept for the rational design of heterogeneous photosensitizers in general and, particularly, for applications in which the antimicrobial activity of singlet oxygen is central.

Effect of concentration on the formation of rose bengal triplet state on microcrystalline cellulose: a combined laser-induced optoacoustic spectroscopy, diffuse reflectance flash photolysis, and luminescence study.

Laser-induced optoacoustic spectroscopy (LIOAS), diffuse reflectance laser flash photolysis (DRLFP), and laser-induced luminescence (LIL) have been applied in conjunction to the determination of triplet state quantum yields of Rose Bengal (RB) supported on microcrystalline cellulose, a strongly light-scattering solid. Among the three used methods, the only one capable of providing absolute triplet quantum yields is LIOAS, but DRLFP and LIL aid in demonstrating that the LIOAS signal arises in fact from the triplet state and confirm the trend found with RB concentration. The coherence found for the three techniques demonstrates the usefulness of the approach. Observed triplet quantum yields are nearly constant within a limited concentration range, after which they decay strongly due to the generation of inactive dye aggregates or energy trapping centers. When quantum yields are divided by the fraction of absorbed light exciting the dye, the quotient falls off steadily with concentration, following the same trend as the observed fluorescence quantum yield. The conditions that maximize triplet formation are determined as a compromise between the rising light absorption and the decrease of quantum yield with RB concentration.

Fluorescence enhancement of a fluorescein derivative upon adsorption on cellulose.

9-[1-(2-Methyl-4-methoxyphenyl)]-6-hydroxy-3H-xanthen-3-one (2-Me-4-OMe TG) is a fluorescein derivative dye whose photophysical properties show a remarkable pH dependence. In aqueous solution the fluorescence quantum yield (Φf) of its anionic species is nearly a hundred times higher than that of its neutral species. Such a large difference in Φf makes 2-Me-4-OMe TG useful as an "on-off" pH indicator. Here we report that adsorption on the surface of microcrystalline cellulose exerts a profound effect upon the photophysical properties of 2-Me-4-OMe TG. On the solid only the dye neutral species is observed and its Φf is 0.31 ± 0.10, which is approximately thirty times higher than the value found for the neutral species in aqueous solution (Φf = 0.01). 2-Me-4-OMe TG and Dabcyl (DB) were co-adsorbed on the surface of microcrystalline cellulose to study the transfer of excitation energy from the former to the latter. In the absence of the dye, the formation of DB aggregates is observed at concentrations greater than 0.34 µmol per gram of cellulose, while in the presence of 2-Me-4-OMe TG the formation of DB aggregates is thoroughly inhibited. The quenching of fluorescence of 2-Me-4-OMe TG by DB reaches efficiencies as high as 90% for the most concentrated samples.

Evidence on dye clustering in the sensitization of TiO2 by aluminum phthalocyanine.

As previous studies have shown, the photocatalytic reduction of Cr(VI) to Cr(III) in the presence of 4-chlorophenol can be carried out efficiently under visible irradiation using TiO2 modified with hydroxoaluminum-tricarboxymonoamide phthalocyanine (AlTCPc) in spite of the high aggregation tendency of the dye. In the present work, photocurrent and absorption spectra of AlTCPc modified TiO2 films are studied together with absorption and fluorescence of the dye in solution as a function of the concentration of the dye to clarify (a) the role of aggregates and the nature of the species responsible for electron injection into the semiconductor and (b) the reasons why, as reported earlier, the photocatalytic activity is nearly independent of dye loading at constant TiO2 mass. Results are consistent with the presence of AlTCPc clusters with similar properties both on the TiO2 surface and in H2O-DMSO solution. The actual photoactive species is the monomeric dye electronically coupled to the semiconductor. Monomer concentration depends only slightly on AlTCPc analytical concentration, in a way similar to surfactant monomers in micellar equilibrium, thus explaining the independence of photocatalytic activity on dye concentration.

Effect of concentration on the photophysics of dyes in light-scattering materials.

Photoactive materials based on dye molecules incorporated into thin films or bulk solids are useful for applications as photosensitization, photocatalysis, solar cell sensitization and fluorescent labeling, among others. In most cases, high concentrations of dyes are desirable to maximize light absorption. Under these circumstances, the proximity of dye molecules leads to the formation of aggregates and statistical traps, which dissipate the excitation energy and lower the population of excited states. The search for enhancement of light collection, avoiding energy wasting requires accounting the photophysical parameters quantitatively, including the determination of quantum yields, complicated by the presence of light scattering when particulate materials are considered. In this work we summarize recent advances on the photophysics of dyes in light-scattering materials, with particular focus on the effect of dye concentration. We show how experimental reflectance, fluorescence and laser-induced optoacoustic spectroscopy data can be used together with theoretical models for the quantitative evaluation of inner filter effects, fluorescence and triplet formation quantum yields and energy transfer efficiencies.

Exploiting electron storage in TiO2 nanoparticles for dark reduction of As(V) by accumulated electrons.

UV irradiation of an ethanolic sol of TiO2 nanoparticles produces a trapped Ti(III) centre and a noticeable Burstein shift. Direct evidence that the accumulated charges can drive the otherwise forbidden reduction of As(V) by conduction band electrons is presented.

Eosin Y triplet state as a probe of spatial heterogeneity in microcrystalline cellulose.

The photophysical behavior of eosin Y adsorbed onto microcrystalline cellulose was evaluated by reflectance spectroscopy, steady-state fluorescence spectroscopy and laser induced time-resolved luminescence. On increasing the concentration of the dye, small changes in absorption spectra, fluorescence redshifts and fluorescence quenching are observed. Changes in absorption spectra point to the occurrence of weak exciton interactions among close-lying dye molecules, whereas fluorescence is affected by reabsorption and excitation energy trapping. Phosphorescence decays are concentration independent as a result of the negligible exciton interaction of dye pairs in the triplet state. Lifetime distribution and bilinear regression analyses of time-resolved phosphorescence and delayed fluorescence spectra reveal the existence of two different environments: long-lived, more energetic triplet states arise from dyes tightly entrapped within the cellulose chains, while short-lived, less-energetic states result from dyes in more flexible environments. Stronger hydrogen bond interactions between the dye and cellulose hydroxyl groups lead in the latter case to a lower triplet energy and faster radiationless decay. These effects, observed also at low temperatures, are similar to those encountered in several amorphous systems, but rather than being originated in changes in the environment during the triplet lifetime, they are ascribed in this case to spatial heterogeneity.

Triplet quantum yields in light-scattering powder samples measured by laser-induced optoacoustic spectroscopy (LIOAS).

In recent years, different methods and techniques have been applied to study the primary photophysical processes occurring in dye-loaded light-scattering powdered samples. In spite of this, there are still no reliable methods for the determination of triplet quantum yields for this kind of systems. Laser-induced optoacoustic spectroscopy (LIOAS) has been extensively used for the determination of triplet quantum yields of dyes in solution. In a previous work, LIOAS was applied to the measurement of absolute emission quantum yields of highly fluorescent powdered samples. Excellent agreement was found with values obtained from reflectance data. In this work, we apply the same technique for the determination of triplet quantum yields of Rose Bengal and Erythrosine B adsorbed on microcrystalline cellulose. In contrast to water and other solvents, internal conversion cannot be neglected in the cellulose environment. The triplet quantum yield for both dyes is around 0.55 and does not change with dye concentration.

Phloxine B as a probe for entrapment in microcrystalline cellulose.

The photophysical behaviour of phloxine B adsorbed onto microcrystalline cellulose was evaluated by reflectance spectroscopy and laser induced time-resolved luminescence in the picosecond-nanosecond and microsecond-millisecond ranges. Analysis of the absorption spectral changes with concentration points to a small tendency of the dye to aggregate in the range of concentrations under study. Prompt fluorescence, phosphorescence and delayed fluorescence spectral decays were measured at room temperature and 77 K, without the need of sample degassing because cellulose protects triplet states from oxygen quenching. In all cases, spectral changes with time and lifetime distribution analysis were consistent with the dye coexisting in two different environments: dyes tightly entrapped between polymer chains in crystalline regions of cellulose showed longer fluorescence and phosphorescence lifetimes and more energetic triplet states, while dyes adsorbed in more amorphous regions of the support showed shorter lifetimes and less energetic triplet states. This behaviour is discussed in terms of the different dye-support interactions in both kinds of adsorption sites.

Silicon nanoparticle photophysics and singlet oxygen generation.

The effect of molecular oxygen and water on the blue photoluminescence of silicon nanoparticles synthesized by anodic oxidation of silicon wafers and surface functionalized with 2-methyl 2-propenoic acid methyl ester is investigated. The particles of 3 +/- 1 nm diameter and a surface composition of Si(3)O(6)(C(5)O(2)H(8)) exhibit room-temperature luminescence in the wavelength range 300-600 nm upon excitation with 300-400 nm light. The luminescence shows vibronic resolution and high quantum yields in toluene suspensions, while a vibronically unresolved spectrum and lower emission quantum yields are observed in aqueous suspensions. The luminescence intensity, though not the spectrum features, depends on the presence of dissolved O(2). Strikingly, the luminescence decay time on the order of 1 ns does not depend on the solvent or on the presence of O(2). To determine the mechanisms involved in these processes, time-resolved and steady-state experiments are performed. These include low-temperature luminescence, heavy atom effect, singlet molecular oxygen ((1)O(2)) phosphorescence detection, reaction of specific probes with (1)O(2), and determination of O(2) and N(2) adsorption isotherms at 77 K. The results obtained indicate that physisorbed O(2) is capable of quenching nondiffusively the particle luminescence at room temperature. The most probable mechanism for (1)O(2) generation involves the energy transfer from an exciton singlet state to O(2) to yield an exciton triplet of low energy (<0.98 eV) and (1)O(2). In aqueous solutions, excited silicon nanoparticles are able to reduce methylviologen on its surface.

Dye-polyelectrolyte layer-by-layer self-assembled materials: molecular aggregation, structural stability, and singlet oxygen photogeneration.

The interaction of rose Bengal (RB) and fluorescein (FL) with poly[diallyldimethylammonium] chloride (PDDA) was studied in layer-by-layer self-assembled thin films and in solution. The spectroscopic behavior is explained in terms of dye-dye, dye-polyelectrolyte, and in solution, dye-solvent interactions. A correlation among dye hydrophobicity, aggregation tendency, polymer folding in solution, and the stability of self-assembled films is obtained. In spite of the very high dye concentration (approximately 1 M), RB-PDDA multilayer thin films are able to photogenerate singlet molecular oxygen, as demonstrated by chemical monitoring and IR phosphorescence detection.

Trapping of Rhodamine 6G excitation energy on cellulose microparticles.

Rhodamine 6G (R6G) was adsorbed on cellulose microparticles and fluorescence quantum yields and decays were measured as a function of dye loading. Though no spectroscopic evidence of dye aggregation was found, a noticeable decrease of quantum yield--after correction for reabsorption and reemission of fluorescence--and shortening of decays were observed at the highest loadings. These effects were attributed to the dissipation of the excitation energy by traps constituted by R6G pairs, leading to static and dynamic quenching produced by direct absorption of traps and non-radiative energy transfer from monomers, respectively. Regarding the nature of traps, two extreme approaches were considered: (a) equilibrium between monomers slightly interacting in the ground state and (b) randomly distributed monomers located below a critical distance (statistical traps). Both approaches accounted quantitatively for the observed facts. The effect of energy migration was evaluated through computational simulations. As the concentration of traps could only be indirectly inferred, in some experiments an external energy transfer quencher, Methylene Blue, was coadsorbed and the results were compared with those obtained with pure R6G.

Validation of fluorescence quantum yields for light-scattering powdered samples by laser-induced optoacoustic spectroscopy.

Determination of quantum yields for various processes (such as fluorescence and triplet formation) in dye-loaded light-scattering powdered samples is an open issue. Here, we report the testing of laser-induced optoacoustic spectroscopy (LIOAS) for the determination of fluorescence quantum yields of Rhodamine 101 and Rhodamine 6G adsorbed at various loadings on microgranular cellulose powder. The results of the LIOAS experiments are consistent with those from the method developed in one of our laboratories based on the measurement of apparent reflectance using an integrating sphere [Mirenda, M.; Lagorio, M. G.; San Roman, E. Langmuir 2004, 20, 3690-3697], which allows the simultaneous calculation of fluorescence quantum yield and reflectance devoid of fluorescence artifacts. Criteria to quantify overall errors and detect outlying values are developed. The theory underlying the application of LIOAS to light-scattering samples and experimental details are presented.

Effect of molecular interactions on the photophysics of Rose Bengal in polyelectrolyte solutions and self-assembled thin films.

Solutions and layer-by-layer self-assembled thin films containing Rose Bengal and poly(diallyldimethylammonium chloride) are studied with the aim of understanding the interactions controlling their structures and the photophysics of the dye in both media. A detailed spectroscopic and theoretical analysis shows that hydrophobic interactions among dye molecules contribute to the coiling of the polyelectrolyte chain in solution at low polyelectrolyte/dye ( P/ D) ratios, whereas extensive aggregation of the dye takes place even at ratios as high as 10(4) (expressed in monomeric units). A polyelectrolyte elongated form prevails in self-assembled thin films, providing an environment that reduces hydrophobic interactions and lowers the aggregation tendency. Self-assembled films with a roughly estimated overall dye concentration around 1 M at a P/ D ratio in the order of seven are fluorescent and photogenerate singlet molecular oxygen. This contrasts with the behavior of polyelectrolyte solutions, which are almost nonfluorescent and do not evidence triplet state generation at the same P/ D ratio.

Excitation energy transfer and trapping in dye-loaded solid particles.

The photophysics of several systems composed of a single dye or pairs of dyes attached to solid particles has been studied in the dry solid state at high dye concentrations taking into account light scattering and inner filter effects. Interaction among dye molecules and singlet-singlet energy transfer are relevant in these conditions, as has been demonstrated for pairs of dyes with suitable spectral overlap. For single dyes, after correction for radiative energy transfer, fluorescence quenching is observed as the surface concentration increases. This effect is explained by two different trapping models. Irrespective of the nature of the traps, concentration quenching may be of static (trap absorption) and dynamic (energy transfer) nature. The unraveling of energy trapping mechanisms is a key to the development of efficient photoactive solid materials.

Contaminación sobre ruedas

Los autores plantean que el tránsito automotor es la mayor fuente de contaminación de la atmósfera de la ciudad de Buenos Aires. El INQUIMAE (Instituto de Química Física de los Materiales, Medio Ambiente y Energía) da un panorama de los distintos gases que intervienen en la contaminación del transporte automotor y su influencia en la calidad del aire y el ser humano