RESUMEN
Fluorescence microscopy is limited by photoconversion due to continuous illumination, which results in not only photobleaching but also conversion of fluorescent molecules into species of different spectral properties through photoblueing. Here, we determined different fluorescence parameters of photoconverted products for various fluorophores under standard confocal and stimulated emission depletion (STED) microscopy conditions. We observed changes in both fluorescence spectra and lifetimes that can cause artifacts in quantitative measurements, which can be avoided by using exchangeable dyes.
Asunto(s)
Artefactos , Microscopía Confocal , Microscopía Fluorescente , Microscopía Fluorescente/métodos , Microscopía Confocal/métodos , Colorantes Fluorescentes/química , FotoblanqueoRESUMEN
Optical tissue clearing and three-dimensional (3D) immunofluorescence (IF) microscopy is transforming imaging of the complex tumor microenvironment (TME). However, current 3D IF microscopy has restricted multiplexity; only 3 or 4 cellular and noncellular TME components can be localized in cleared tumor tissue. Here we report a light-emitting diode (LED) photobleaching method and its application for 3D multiplexed optical mapping of the TME. We built a high-power LED light irradiation device and temperature-controlled chamber for completely bleaching fluorescent signals throughout optically cleared tumor tissues without compromise of tissue and protein antigen integrity. With newly developed tissue mounting and selected region-tracking methods, we established a cyclic workflow involving IF staining, tissue clearing, 3D confocal microscopy, and LED photobleaching. By registering microscope channel images generated through 3 work cycles, we produced 8-plex image data from individual 400 µm-thick tumor macrosections that visualize various vascular, immune, and cancer cells in the same TME at tissue-wide and cellular levels in 3D. Our method was also validated for quantitative 3D spatial analysis of cellular remodeling in the TME after immunotherapy. These results demonstrate that our LED photobleaching system and its workflow offer a novel approach to increase the multiplexing power of 3D IF microscopy for studying tumor heterogeneity and response to therapy.
Asunto(s)
Imagenología Tridimensional , Microscopía Fluorescente , Fotoblanqueo , Microambiente Tumoral , Animales , Imagenología Tridimensional/métodos , Microscopía Fluorescente/métodos , Ratones , Humanos , Línea Celular Tumoral , Microscopía Confocal/métodos , FemeninoRESUMEN
Fluorescence recovery after photobleaching (FRAP) is now an indispensable tool to analyze the diffusion of molecules in vivo and in vitro. However, a conventional fluorescence intensity-based approach has difficulty in analyzing the diffusion of multiple species simultaneously. Here, we report fluorescence lifetime recovery after photobleaching (FLRAP) that incorporates fluorescence lifetime information into FRAP. By using FLRAP, the fluorescence intensity-recovery curves of each species can be successfully extracted from the ensemble photon data by utilizing their species-specific fluorescence decay curves, which are verified by applying FLRAP to two heterogeneous systems. Thus, FLRAP can be a powerful tool to quantitatively elucidate the molecular diffusion of multiple species in complex systems such as in living cells.
Asunto(s)
Fotones , Recuperación de Fluorescencia tras Fotoblanqueo , Fotoblanqueo , DifusiónRESUMEN
Significance: Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful and versatile imaging technique renowned for its remarkable features, including high-speed 3D tomography, minimal photobleaching, and low phototoxicity. The interference light-sheet fluorescence microscope, with its larger field of view (FOV) and more uniform axial resolution, possesses significant potential for a wide range of applications in biology and medicine. Aim: The aim of this study is to investigate the interference behavior among multiple light sheets (LSs) in LSFM and optimize the FOV and resolution of the light-sheet fluorescence microscope. Approach: We conducted a detailed investigation of the interference effects among LSs through theoretical derivation and numerical simulations, aiming to find optimal parameters. Subsequently, we constructed a customized system of multi-LSFM that incorporates both interference light sheets (ILS) and noninterference light-sheet configurations. We performed beam imaging and microsphere imaging tests to evaluate the FOV and axial resolution of these systems. Results: Using our custom-designed light-sheet fluorescence microscope, we captured the intensity distribution profiles of both interference and noninterference light sheets (NILS). Additionally, we conducted imaging tests on microspheres to assess their imaging outcomes. The ILS not only exhibits a larger FOV compared to the NILS but also demonstrates a more uniform axial resolution. Conclusions: By effectively modulating the interference among multiple LSs, it is possible to optimize the intensity distribution of the LSs, expand the FOV, and achieve a more uniform axial resolution.
Asunto(s)
Microscopía Fluorescente , Microscopía Fluorescente/métodos , Microesferas , FotoblanqueoRESUMEN
The oligomerization of proteins is an important biological control mechanism and has several functions in activity and stability of enzymes, structural proteins, ion channels and transcription factors. The determination of the relevant oligomeric states in terms of geometry (spatial extent), oligomer size (monomer or dimer or oligomer) and affinity (amounts of monomer, dimer and oligomer) is a challenging biophysical problem. Förster resonance energy transfer and fluorescence fluctuation spectroscopy are powerful tools that are sensitive to proximity and oligomerization respectively. Here it is proposed to combine image-based lifetime-detected Forster resonance energy transfer with image correlation spectroscopy and photobleaching to determine distances, oligomer sizes and oligomer distributions. Simulations for simple oligomeric forms illustrate the potential to improve the discrimination between different quaternary states in the cellular milieu.
Asunto(s)
Transferencia Resonante de Energía de Fluorescencia , Fotoblanqueo , Transferencia Resonante de Energía de Fluorescencia/métodos , Multimerización de Proteína , Estructura Cuaternaria de Proteína , Humanos , Simulación por ComputadorRESUMEN
Fluorescent dyes synergize with advanced microscopy for researchers to investigate the location and dynamic processes of biomacromolecules with high spatial and temporal resolution. However, the instability of fluorescent dyes, including photobleaching and photoconversion, represent fundamental limits for super-resolution and time-lapse imaging. In this review, we discuss the latest advances in improving the photostability of fluorescent dyes. We summarize the primary photobleaching processes of cyanine and rhodamine dyes and highlight a range of strategies developed in recent years to strengthen these fluorophores. Additionally, we discuss the influence of protein microenvironments and labeling methods on the photostability of fluorophores. We aim to inspire next-generation robust and bright fluorophores that ultimately enable the routine practice of time-lapse super-resolution imaging of live cells.
Asunto(s)
Colorantes Fluorescentes , Proteínas , Colorantes Fluorescentes/metabolismo , Fotoblanqueo , Microscopía Fluorescente/métodos , RodaminasRESUMEN
Dye-encoded bead-based assays are widely used for diagnostics. Multiple bead populations are required for multiplexing and can be produced using different dye colors, labeling levels, or combinations of dye ratios. Ready-to-use multiplex bead populations restrict users to specific targets, are costly, or require specialized instrumentation. In-house methods produce few bead plexes or require many fine-tuning steps. To expand bead encoding strategies, we present a simple, safe, and cost-effective bench-top system for generating bead populations using photobleaching. By photobleaching commercially available dye-encoded magnetic beads for different durations, we produce three times as many differentiable bead populations on flow cytometry from a single dye color. Our photobleaching system uses a high-power LED module connected to a light concentrator and a heat sink. The beads are photobleached in solution homogeneously by constant mixing. We demonstrate this photobleaching method can be utilized for cross-testing antibodies, which is the first step in developing immunoassays. The assay uses multiple photobleached encoded beads conjugated with capture antibodies to test many binding pairs simultaneously. To further expand the number of antibodies that can be tested at once, several antibodies were conjugated to the same bead, forming a pooled assay. Our assay predicts the performance of antibody pairs used in ultrasensitive Simoa assays, narrowing the number of cross-tested pairs that need to be tested by at least two-thirds and, therefore, providing a rapid alternative for an initial antibody pair screening. The photobleaching system can be utilized for other applications, such as multiplexing, and for photobleaching other particles in solution.
Asunto(s)
Anticuerpos , Fotoblanqueo , Anticuerpos/inmunología , Anticuerpos/química , Microesferas , Citometría de Flujo , Humanos , Inmunoensayo/métodos , Inmunoensayo/instrumentación , Colorantes Fluorescentes/químicaRESUMEN
Advanced 3D high-resolution imaging techniques are essential for investigating biological challenges, such as neural circuit analysis and tumor microenvironment in intact tissues. However, the fluorescence signal emitted by endogenous fluorescent proteins in cleared or expanded biological samples gradually diminishes with repeated irradiation and prolonged imaging, compromising its ability to accurately depict the underlying scientific problem. We have developed a strategy to preserve fluorescence in cleared and expanded tissue samples during prolonged high-resolution three-dimensional imaging. We evaluated various compounds at different concentrations to determine their ability to enhance fluorescence intensity and resistance to photobleaching while maintaining the structural integrity of the tissue. Specifically, we investigated the impact of EDTP utilization on GFP, as it has been observed to significantly improve fluorescence intensity, resistance to photobleaching, and maintain fluorescence during extended room temperature storage. This breakthrough will facilitate extended hydrophilic and hydrogel-based clearing and expansion methods for achieving long-term high-resolution 3D imaging of cleared biological tissues by effectively safeguarding fluorescent proteins within the tissue.
Asunto(s)
Proteínas Fluorescentes Verdes , Imagenología Tridimensional , Proteínas Fluorescentes Verdes/metabolismo , Animales , Imagenología Tridimensional/métodos , Ratones , Fotoblanqueo , FluorescenciaRESUMEN
Abstract The illicit market of counterfeit medicines containing sildenafil and tadalafil has been causing serious public health problems. Thus, further studies on this illicit association are needed. A stability-indicating HPLC method was developed for simultaneous determination of tadalafil (TAD) and sildenafil (SIL) using a C18 column (250 x 4.6 mm, 5 µm). Detection was achieved at 284 nm, for TAD, and 292 nm, for SIL. The method was considered to be specific, linear, precise, accurate, robust, and sensitive. In the photodegradation kinetic studies, the drugs showed a first-order reaction rate when isolated, and zero-order when associated. Toxicological assays demonstrated that the photodegraded drugs decreased cell viability in compared to non- degraded drugs, suggesting cytotoxic activity. Additional, mutagenic activity was not observed under the tested conditions. Photodegraded drugs, in association, depicted DNA damage index, suggesting genotoxic effects. The obtained results will be able to support the forensic intelligence laboratories, as well as to alert the population about the risk inherent to consuming counterfeit products.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Fotoblanqueo/efectos de los fármacos , Citrato de Sildenafil/análisis , Tadalafilo/análisis , Medicamentos Falsificados/clasificaciónRESUMEN
Light sheet microscopy (LSM) is an evolving optical imaging technique with a plane illumination for optical sectioning and volumetric imaging spanning cell biology, embryology, and in vivo live imaging. Here, we focus on emerging biomedical applications of LSM for tissue samples. Decoupling of the light sheet illumination from detection enables high-speed and large field-of-view imaging with minimal photobleaching and phototoxicity. These unique characteristics of the LSM technique can be easily adapted and potentially replace conventional histopathological procedures. In this review, we cover LSM technology from its inception to its most advanced technology; in particular, we highlight the human histopathological imaging applications to demonstrate LSM's rapid diagnostic ability in comparison with conventional histopathological procedures. We anticipate that the LSM technique can become a useful three-dimensional imaging tool for assessing human biopsies in the near future.
Asunto(s)
Humanos , Biopsia , Dermatitis Fototóxica , Embriología , Imagenología Tridimensional , Iluminación , Microscopía , Imagen Óptica , FotoblanqueoRESUMEN
A simple and fast alternative methodology using capillary zone electrophoresis (CZE) to analyze linezolid and its cationic photodegradation products in tablets has been developed. The separation was carried out on fused silica capillary and conducted using 100 mM formic acid (pH 3.0) and by applying 30 KV voltage. Detection was performed at UV 254 nm. The optimized method was validated in terms of linearity, limits of detection and quantification, precision (repeatability), stability studies (selectivity) and accuracy. Good linearity (8-20 mg L-1) was obtained and the limit of detection was 0.95 mg L-1. The greatest advantages of the CZE method were the rapid set-up of instrumentation and capillary equilibration, short analysis time (12 min), low running cost and low waste generation. The method showed good stability in determining linezolid submitted to degradation by light and to a climatic chamber and can be used as an alternative for evaluation in stability studies of linezolid in tablets, as well as for the analysis of the drug in raw materials and finished products.
Asunto(s)
Comprimidos/clasificación , Electroforesis Capilar/instrumentación , Linezolid/análisis , Fotoblanqueo , Medicamentos del Componente Especializado de los Servicios FarmacéuticosRESUMEN
Dissolved organic carbon (DOC) is nowadays recognized as the main substrate and source of energy for aquatic microbial community. The great part of available organic carbon for bacterioplankton might be formed after photolytic degradation of humic material, which constitutes the major part of DOC in almost all natural waters. The effects of DOC photo-degradation were evaluated, as was its utilization by bacterioplankton, through a two-step experiment, one involving photo-degradation of DOC and the other bacterial growth on the photo-degraded substrate. Photo-degradation was responsible for the consumption of 19% of DOC, reduced SUVA254, an increase in the E2/E3 and E3/E4 ratios, in addition to modifications in the fluorescence spectra that indicated a rise in the labile fraction of DOC. However, these alterations on DOC were not reflected in differences in bacterioplankton growth, as shown by the fact that there were no significant differences in density, biomass, bacterial production, bacterial respiration and bacterial growth efficiency between treatment and control.
O carbono orgânico dissolvido (COD) é reconhecido atualmente como o principal substrato e fonte de energia para a comunidade microbiana aquática. Grande parte do COD biodisponível para o bacterioplâncton pode ser gerada após degradação fotolítica do material húmico dissolvido, que constitui a maior parte do carbono orgânico total. Neste trabalho foram avaliados os efeitos da fotodegração no COD de um ambiente húmico e a sua posterior utilização pelo bacterioplâncton, por meio de um experimento em duas etapas, uma de fotodegradação do COD e outra de crescimento microbiano sobre o substrato fotodegradado. A fotodegradação causou consumo de 19% do COD, diminuição na SUVA254, aumento nas razões E2/E3 e E3/E4 e modificações nos espectros de fluorescência apontando aumento da fração lábil. No entanto, as alterações no COD não se refletiram em diferenças no crescimento do bacterioplâncton, não havendo diferenças significativas com relação à densidade, biomassa, produção, respiração e eficiência de crescimento bacteriano.
Asunto(s)
Carbono , FotoblanqueoRESUMEN
A stability study of azithromycin in ophthalmic preparations was developed by submission to different types of light, temperature and pH, using the biodiffusion assay (cylinder 3 x 3) for the quantifications. Bacillus subtilis, ATCC 9372, was used as test organism. The used concentration range was of 50 to 200 µg/mL. The study demonstrated that the drug suffered degradation when submitted to the ultraviolet light, germicide light, solar luminosity, acid solution, basic solution and hydrogen peroxide solution. The results were analyzed by the analysis of variance (ANOVA).
O estudo de estabilidade de azitromicina em preparações oftálmicas foi realizado após exposição a diferentes tipos de luz, temperatura e pH, utilizando o método de difusão em ágar (cilindros 3 x 3) para as quantificações. A faixa de concentração foi de 50 a 200 µg/mL. O estudo demonstrou que o fármaco sofreu degradação quando submetido às luzes ultravioleta, germicida e solar, e a soluções ácida, alcalina e de peróxido de hidrogênio. Os resultados foram analisados através da análise da variância (ANOVA).
Asunto(s)
Evaluación de Medicamentos/estadística & datos numéricos , Azitromicina/farmacología , Oftalmología , Preparaciones Farmacéuticas , Análisis de Varianza , Bioensayo , Difusión Facilitada , FotoblanqueoRESUMEN
O objetivo do presente trabalho foi estudar a degradação fotocatalítica de três corantes e um interferente endócrino mediado pelo dióxido de titânio (TiO2). Foi estudada a degradação fotocatalítica de dois azo corantes comerciais, respectivamente o C.I Reactive Black 5 e o C.I Reactive Red 239. Para esse estudo foi utilizado o TiO2 P25 Evonik como catalisador e os experimentos foram realizados em solução aquosa com reator artificial equipado com lâmpada de vapor de mercúrio 125W. Nesse estudo,foram avaliados os seguintes parâmetros: concentração de TiO2, tempo de irradiação, pH, concentração inicial de corante, presença de diferentes concentrações de peróxido de hidrogênio e degradação em sistema bi-compartimental para os dois azo corantes. O outro corante estudado foi o Índigo Carmine (IC). Primeiramente foi investigada a descoloração do corante em efluente modelo usando processo físico e químico para sua remoção. Os experimentos de descoloração do efluente contendo o corante IC foram realizados usando carvão ativado no escuro e com irradiação. Além disso, o efeito do carvão ativado / TiO2 / UV foi avaliado pelo mesmo sistema. Os parâmetros da degradação do IC foram realizados, seguindo: concentração de TiO2, pH, concentração inicial de corante, presença de ânions inorgânicos, temperatura, concentração de peróxido de hidrogênio. A eficiência de três diferentes reatores (Reator 1 reator do tipo batch com lâmpada vapor de mercúrio 125W; Reator 2 reator do tipo batch equipado com 4 lâmpadas daylight 20W; Reator 3 reator tubular de fluxo contínuo equipado com uma lâmpada daylight 20W) e reator batch com luz solar foram avaliados para degradação do IC.Além disso, otimizados experimentos foram realizados em planta piloto com compound parabolic collector reatores (CPC) na Plataforma Solar de Almería, Espanha, com TiO2 em suspensão e suportado. O IC foi estudado em diferentes tipos de águas, sendo elas: água destilada, água sintética moderadamente dura, água sintética e real de estação de tratamento de água residuária (ETAR). Testes ecotoxicológicos para o IC foram realizados em dois diferentes níveis tróficos de cadeia alimentar (Daphnia similis e Pseudokirchneriella subcapitata) e testes com minhocas Eisenia andrei. A degradação fotocatalítica do Bisfenol A (BPA) em diferentes tipos de águas na presença de TiO2 foi feita sob diferentes condições. O TiO2 em suspensão foi utilizado para comparar a eficiência da degradação do BPA (...) em batch e CPC reatores. O TiO2 foi suportado em esferas de vidro pelo método sol-gel e usado em reatores solares CPC em escala piloto para degradação do BPA (...) A influência das espécies reativas (...) foi avaliada. Testes ecotoxicológicos com minhocas Eisenia andrei também foram realizados para o BPA e seus produtos de degradação.
Asunto(s)
Humanos , Colorantes/toxicidad , Ambiente , Fotoblanqueo , Sistema Endocrino/metabolismo , Titanio , Purificación del Agua , Ecotoxicología/métodos , Industria Textil/tendenciasRESUMEN
Este estudo avaliou a influência do tipo de fonte fotopolimerizadora na resistência ao cisalhamento da união adesiva ao esmalte clareado com peróxido de carbamida a 10%. Setenta e dois dentes incisivos bovinos íntegros foram divididos em 6 grupos (n=12) de acordo com a realização ou não do tratamento clareador e com a fonte fotoativadora empregada: G1 ù clareamento/luz halógena; G2 ù sem clareamento (controle)/luz halógena; G3 ù clareamento/LED; G4 ù sem clareamento (controle)/LED; G5 ù clareamento/arco de plasma de xenônio; G6 ù sem clareamento/arco de plasma de xenônio. Os grupos experimentais foram submetidos ao clareamento por um período de 14 dias, com exposição diária 4 horas. Os grupos controle foram armazenados em saliva artificial, a 37ºC, por período igual ao clareamento. A união da resina composta ao esmalte obedeceu às recomendações dos fabricantes. Em seguida, todos os corpos de prova foram armazenados em água destilada, por 24 horas, a 37ºC, até a realização do ensaio de resistência ao cisalhamento. O maior valor médio de resistência (14,86 MPa) foi obtido para o grupo G2, e o menor para o grupo G5 (12,32 MPa). A comparação dos grupos através da análise de variância (ANOVA 2-way) demonstrou não haver diferenças significativas para ambos os fatores considerados neste estudo (clareamento e tipo de fonte fotopolimerizadora), bem como para a interação entre estes. Sendo assim, foi possível concluir que o clareamento do esmalte bem como o tipo de fonte fotopolimerizadora utilizada, não interferiram na resistência da união adesiva a este substrato.
Asunto(s)
Animales , Bovinos , Blanqueamiento de Dientes/efectos adversos , Fotoblanqueo , Resistencia al Corte , Esmalte Dental , Luz/efectos adversosRESUMEN
Sparfloxacin, a third generation fluoroquinolone derivative, is a potent antibacterial agent active against a wide range of Gram-positive and Gram-negative organisms including Streptococcus pneumoniae, Staphylococcus aureus, methicillin resistant S. aureus, Legionella spp., Mycoplasma spp., Chlamydia spp. and Mycobacterium spp. A drawback of fluoroquinolones is their photoreactivity. Sparfloxacin has been studied in terms of therapeutic activities. However, there are few published of analytical methods being applied to sparfloxacin. The aim in this study was to determine the photodegradation products of sparfloxacin, when submitted to UV light, and to characterize two of these products, designated SPAX-PDP1 and SPAX-PDP2. An accelerated study of stability in methanol solution was carried out by exposing a solution of sparfloxacin to UV light (peak wavelength 290 nm) for 36 hours at room temperature. The products were analyzed by NMR spectrophotometry, IR spectrometry and mass spectrophotometry. The results suggest that the products isolated here could be used to estimate the degradation of sparfloxacin in a stability study. However, the low activity exhibited by UV-irradiated sparfloxacin is a source of concern that demands further investigation of the mechanism of its photodegradation mechanism
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Fluoroquinolonas/aislamiento & purificación , Fotoblanqueo , Estabilidad de Medicamentos , Luz , Control de CalidadRESUMEN
O objetivo deste trabalho foi avaliar o tempo de utilização das soluções processadoras e quantidade de filmes radiográficos que podem ser processados nessas mesmas soluções sem serem trocadas antes que ocorra degradação e exaustão desses líquidos. O método de avaliação foi a análise subjetiva da qualidade das imagens readiográficas. Para tanto, foram utilizados 269 filmes periapicais Ektaspeed Plus Kodak, do grupo E, quanto à sensibilidade, divididos em dois grupos: degradação e exaustão. Os filmes foram processados em câmara escura portátil sob luz ambiente de lâmpada fluorescentes para simular condições normais de trabalho em consultório odontológico. Após o processamento e secagem os filmes foram avaliados por cinco alunos do curso de pós-graduação em Radiologia Odontológica da FOSJC - Unesp pelo método subjetivo por meio de notas. Pôde-se constatar que para mo grupo degradação as piores notas (K < 1) foram emitidas a partir da película número 43; para o grupo exautão as piores notas foram atribuídas a partir da 192º radiografia (K = 1,8), porém neste grupo as radiografias compreendidas entre as de números 192 até 210 foram classificadas como de boa qualidade para interpretação. Conclui-se que a degradação do revelador ocorreu a partir do 21º dia do inicio do experimento, observado na 43ª radiografia, e, para o grupo exaustão,a partir da 192ª radiografia revelada, que ocorreu no nono dia da pesquisa
Asunto(s)
Fotoblanqueo , Radiografía Dental , Soluciones/análisis , Película para Rayos XRESUMEN
As propriedades fotofísicas e fotoquímicas de cristal violeta (CV) foram investigadas em soluções isotrópicas e verificou-se que solventes com constante dielétrica pequena favorecem a formação do par iônico, já o aumento na viscosidade do meio restringe a movimentação rotacional dos anéis aromáticos, resultando em diminuição da conversão interna e conseqüentemente. aumento na fluorescência e, portanto no rendimento quântico de fluorescência (fi IND. f) (Oliveira 2002). Os experimentos com CV foram conduzidos em micelas reversas do anfifílico aniônico bis-2-etilhexil sulfoccinato de sódio (AOT) em isooctano. A localização interfacial do CV nas micelas reversas de AOT em valores da razão molar entre água e surfactante (W IND. 0) pequenos e grandes foram encontrados através da técnica de Ressonância Magnética Nuclear (RMN) de próton e de carbono 13. Utilizando-se espectroscopia UV-Vis identificou-se que pares iônicos de contato estão presentes a valores pequenos de W IND. 0 e com o aumento do W IND. 0 pares iônicos separados por solventes são as espécies que predominam em solução. A comparação da eficiência de fotodegradação de CV em micelas reversas de AOT em função do W IND. 0 indicou que a fotoreatividade é maior em baixos valores de W IND. 0. Este efeito deve estar relacionado à restrição da movimentação dos anéis aromáticos de CV devido ao ambiente no qual este se localiza na micela reversa de AOT a W IND. 0 pequenos. A formação de intermediários reativos foi verificada através de Fotólise de Relâmpago a Laser e Emissão no infra-vermelho próximo, indicando a presença de espécies triplete, radical e oxigênio singlete com valor de rendimento quântico menor que 1%...