A new laser dye with potential for high stability and a broad band of lasing action: Perylene-3,4,9,10-tetracarboxylic acid-bis-N,N′(2′,6′ xylidyl)diimide

We have obtained high gain lasing action from a dye in the calss of perylene-tetracarboxylic acid diimides. These dyes have been used in the past as microcrystalline pigments and more recently as molecularly dissolved fluors having both an orange absorption color and orange fluorescence. In dimethylformamide this dye absorbs with a main maximum at 527 nm, well coupled to the exciting radiation from a frequency-doubled YAG exciting laser. The dye lases primarily in the 0′→ 1″ satellite of the fluorescence band from 566–585 nm, with intermittent output pulses to 605 nm. While this dye overlaps the lasing bands of the rhodamine dyes, we believe that it may be considerably more stable than even Rhodamine 6G, the most stable of that class.     

Mechanistic studies of oxidation reactions by fluorescence spectroscopy: a critical assessment

The synthesis and properties of a ratiometric fluorescent dye system as a probe for the detection of oxidizing species are described. The dye consists of a strongly fluorescent Bodipy label attached to a styryl moiety. The oxidation of the exocyclic double bond by agents for syn‐ and anti‐ dihydroxylation leads to a visible change in fluorescence colour from orange to green. IR spectroscopy provides evidence for the generation of the epoxide in the reaction with peroxides, whereas a ketone and a product of unknown structure are produced in the reaction with osmiumtetroxide, each in contrast to the expected hydroxylations. Based on these first attempts, we discuss the significance of fluorescence spectroscopy to track chemical reactions at low concentrations and give a perspective of how present shortcomings can be overcome. Copyright © 2009 John Wiley & Sons, Ltd.     

Rutacridone as a Fluorescent Dye for the Study of Pollen

Rutacridone, alkaloid from plant Ruta graveolens was used for the histochemical staining of Hippeastrum hybridum pollen. The rutacridone fluorescence (excited by UV light at 360–380 nm) has been studied with Karnaukhov-constructed microspectrofluorimeters. Nucleus and DNA were stained by this dye in cells and fluoresced by green light with maxima of 530 and 555 nm, respectively. Development of the pollen was analyzed with this dye.     

Fluorescence Study of DNA–Dye Complexes Using One-Photon and Two-Photon Picosecond Excitation

Preliminary results of investigation of one-photon- and two-photon-induced fluorescence of acridine orange (AO), epirubicin (ER), hypericin (HYP), and ethidium bromide (EB) in complexes with DNA are presented. A spectrofluorometer based on a picosecond Nd:YAG laser was used for investigations of two-photon (1064-nm, 1-mJ, 40-ps) and one-photon (532- and 355-nm) dye excitation. The spectra of two-photon-induced fluorescence of dyes and their complexes with DNA as well as the kinetics of dyes' fluorescence intensification during their interactions with DNA in dependence on the biomacromolecule concentration were obtained. The intensities of AO, HYP, and EB fluorescence were increased 2.4, 3.2, and 8 times, respectively, after binding with DNA at two-photon excitation, while at one-photon excitation the corresponding values were 2.5, 3.7, and 10 times. The difference in fluorescence enhancement during DNA–dye complex formation at linear and nonlinear excitation may possibly be associated with the fact that the cross sections of one-photon and two-photon absorption, in general, change unequally during the binding of dyes to organic molecules and bathocromic shift of the electronic transitions. It was shown that the peak of AO fluorescence shifted to a longer wavelength on 10 nm after two-photon excitation at 1064 nm in comparison with one-photon excitation at 532 nm.     

Fluorescence and Laser Emission from Coumarin-Acridine Orange Mixtures

The sensitized fluorescence and laser emissions of dye mixtures; (I) coumarin 102 (donor) and acridine orange (acceptor) and (II) coumarin 47 (donor) and acridine orange (acceptor) with Hg-lamp and N₂ laser, have been measured as a function of dye concentration and of the pump power (N₂ laser). Acridine orange which does not lase by itself on excitation with N₂ laser, lases efficiently in the presence of 7-amino-coumarins via singlet-singlet energy transfer. Energy transfer rate constants and critical distances have been estimated from fluorescence intensity and lifetime measurements. The performance of energy transfer dye lasers (ETDLs) are discussed in terms of spectral characteristics of the dyes and their penetration depths.     

Fluorescent polystyrene microspheres with large Stokes shift by fluorescence resonance energy transfer

We prepared fluorescent microspheres with notably large Stokes shift and long-wavelength fluorescence by applying fluorescence resonance energy transfer (FRET) between two common julolidine dyes. Short distance between dye molecules caused by high dye concentration results in efficient FRET in microspheres. However, adequate dye concentration and moderate molar ratio of the donor and acceptor should be chosen to avoid aggregation of dye molecules, which leads to the decrease of fluorescent intensity. Microrspheres with average distance between dye molecules of 1.94 nm and molar ratio of 3.08:1 realize highly efficient FRET with no fluorescence of donor and intense long-wavelength emission of acceptor. In addition, the applied solvent evaporation method for preparing microspheres provided better protection of dyes from ambient medium than traditional surface-labeled method. These results demonstrate the feasibility of applying FRET in microspheres to expand useful fluorescent probes, and reveal their potential application in bioassays field.     

The Biarsenical Dye Lumio™ Exhibits a Reduced Ability to Specifically Detect Tetracysteine-Containing Proteins Within Live Cells

Investigating the localisation of proteins within live cells via fluorescence microscopy typically involves the fusion of the protein of interest to a large fluorescent protein such as green fluorescent protein (GFP). Alternate fluorescent labelling technologies such as the fluorescent biarsenical dye molecules (e.g. FlAsH, ReAsH) are preferable to the use of large fusion proteins in many respects and allow greater flexibility in terms of the location of the labelling site. We assessed the ability of the FlAsH-derived biarsenical dye molecule Lumio™ to label a range of tetracysteine containing proteins within live cells and report that although in some circumstances Lumio is capable of positively detecting such proteins, the sensitivity and specificity of labelling is significantly reduced, making the Lumio-labelling system unsuitable for the detection of a wide range of protein within live cells.     

Fluorescence Study of Excited-State Relaxation Processes of 2-Pyridyl-5-Aryloxazoles

Anomalously high fluorescence spectrum Stokes' shifts of 2(4-pyridyl)-5-(4-methylphenyl)oxazole, 2(4-pyridyl)-5-(4-methoxyphenyl)oxazole, 2(4-pyridyl)-5-(4-N,N-dimethylaminophenyl)oxazole have been found. The fluorescence spectra of the compounds studied are shifted to longer wavelengths as the solvent dielectric constant increases. The dipole moments of these compounds increase by 3–4 times upon excitation. The fluorescence spectra of the compounds investigated are shifted to shorter wavelengths, and the fluorescence quantum yields increase as the temperature decreases. The fluorescence rate constant of 2(4-pyridyl)-5-(4-N,N-dimethylaminophenyl)oxazole changes slightly as the temperature decreases from 293 K (relaxed state) to 114 K (mainly nonrelaxed state). It was concluded that the anomalously high fluorescence Stokes' shift of 2-pyridyl-5-aryloxazole derivatives is caused by solvent orientation relaxation.     

橙汁饮品的荧光光谱特性及模式识别

应用FLS920型荧光光谱仪测得鲜榨橙汁、市售橙汁饮品(8种品牌)的三维荧光光谱,通过分析鲜榨橙汁与饮品中常见食品添加剂纯物质的荧光机理与特性,得到橙汁饮品中橙汁成分的最佳激发波长为470 nm,荧光峰值波长为526 nm,其荧光特征峰为470/526 nm,其余荧光峰的归属均为食品添加剂。同时,采用三维荧光光谱重心法与基于MCD估计的稳健马氏距离相结合的方法对市售橙汁饮品进行筛选与分类,分类效果良好。结果表明:将荧光光谱技术与光谱模式识别方法相结合,可为饮料类食品的鉴别提供帮助。     

Molecular ruler based on concurrent measurements of enhanced Raman scattering and fluorescence

We report a way to assess the separation between a molecule and a metal surface on account of both Raman scattering and fluorescence intensities measured simultaneously. This approach has no need for an accurate quantity of molecules and bears 1 nm resolution. Its distance sensitivity is experimentally demonstrated in the case of dye molecules lying on a gold surface with a polymer spacer layer in between and is compared with theoretical prediction.     

荧光光谱检测的酸性橙Ⅱ的研究

酸性橙Ⅱ在食品工业中是禁用添加剂.针对酸性橙Ⅱ的食品安全检测问题,报道了运用荧光光谱技术检测其水溶液的实验.实验表明,酸性橙Ⅱ水溶液在230～290nm波长激发下,310～390nm范围内产生荧光光谱,峰位波长在350nm,最佳激励波长为250nm;采用垂直偏振片(起偏角为0°)起偏照射样品,发现偏振荧光峰位不变,荧光峰强度随检偏角的增大而呈明显的线性递减关系.由实验数据计算得到荧光偏振度为0.783,表明分子具有一定确定取向;另外,分析认为酸性橙Ⅱ产生荧光,是由于分子中含有苯环和萘环结构吸收紫外光能量,以及氮键在光子作用下形成顺式异构体的激发单线态后,两者发射的光子所致.整个结果对酸性橙Ⅱ在食品中的违禁使用检测、特性表征、以及分子规律的更深入研究,有一定的参考价值.     

Generation of nearly transform-limited pulses in subnanosecond region with TEA nitrogen-laser-pumped dye laser

A dye laser is transversely pumped by a TEA nitrogen laser with 700 ps duration. The dye laser pulse has a spectral linewidth of 0.007 nm and a pulse width of ≈ 140 ps, giving a time-bandwidth product of 1.5. It is pointed out that the fluorescence lifetime of the laser dye plays an important role in generation of laser emission using a long laser cavity under subnanosecond pulse pumping.     

Measurement of two-photon excitation cross-section of a new symmetric-type fluorene derivative

A new dye, 2,7-bis(4-methoxystyryl)-9,9-bis(2-ethylhexyl)-9H-fluorene, has been synthesized, which is a d-π-d symmetrical-type fluorene derivative. The two-photon absorption (TPA) of this new dye has been experimentally studied by comparable two-photon-induced fluorescence method. This new dye has a TPA cross-section of at 790 nm/13 fs.     

Synthesis and Characterization of a Fluorescent Water-Borne Polyurethane Based on a Novel Naphthalimide Dye

A waterborne-polyurethane (WPU) dye, based on the fluorescent dye 4-diamino propane-N-allyl-1,8- naphthalimide (WPU-DAN), was synthesized by attaching 4-diamino propane-N-allyl-1,8- naphthalimide (DAN) onto both ends of the polyurethane (PU) chains according to a prepolymer?ionomer process. The synthetic process was confirmed by Fourier transform infrared spectroscopy (FTIR) and UV-visible absorption. The glass transition temperature, molecular weight and average particle sizes were measured. The glass transition temperatures of WPU and WPU-DAN were 46.6 and 49.8°C, respectively. In addition, the particle size distributions of WPU-DAN and WPU were 140 and 134 nm, respectively. The thermal behaviour of WPU-DAN showed improvement compared to WPU. The fluorescence intensity of WPU-DAN was enhanced more than DAN due to the naphthalimide groups attached to the chains, and the fluorescence intensity of WPU-DAN and DAN were increased by increasing temperature. Moreover, the fluorescence intensity of WPU-DAN emulsion was stable during 30 days and no loss of fluorescence intensity occurred for these days.     

Synthesis of high fluorescent silica hybrid materials by immobilization of orange peel extract in silica‐silsesquioxane matrix

Sol–gel chemistry can be easily modified to the changing needs of society to produce fine‐tuned sol–gel nanostructured materials for relevant applications. In this context, there is an increasing need for natural and versatile raw materials as well as biocompatible compounds that could be extensively used as biomarkers for bioimaging in diagnosis or therapy. Silica‐based materials are widely used in the field of biomedicine due to their chemical inertness and biocompatibility. In the present paper, orange peel extract was immobilized inside inorganic silica and hybrid silica‐silsesquioxane polymeric networks. Silica and organo‐modified silica matrices were synthesized through a templated sol–gel route of TEOS and an organosilsesquioxane (octaisobutyltetracyclo[7.3.3.1^5,11] octasiloxane‐endo‐3,7‐diol), with D ‐glucose as template, and for comparison a non‐ionic surfactant (tetraethylene glycol mono‐hexadecyl ether) was also used. The bioactive properties of the molecules from orange peel extract were preserved after immobilization in both silica and silica‐silsesquioxane networks. Moreover, the fluorescence properties were amplified by 10–20 times more than the native orange peel extract. The structural properties of the final materials have been studied by FT‐IR, UV–Vis‐NIR, and fluorescence spectroscopy. Dynamic light scattering measurements and transmission electron microscopy were used to estimate the size and morphology of the hybrid materials with orange peel extract immobilized in silica networks. Copyright © 2009 John Wiley & Sons, Ltd.     

Fluorescence Analysis of Acridine Orange Adsorbate at the Water/N-Heptane Interface, Bulk and Interface

The amphiphilic nature of 3, 6-bisdimethylaminoacridine (trivial name, acridine orange) can be used to characterize (polar/nonpolar) interfaces. This paper studies the properties of acridine orange and its dodecyl derivative at a solvent interface by internal reflection and tension determinations. In the case of absorbance measurements of crystallized acridine orange, the potenz value of the conventional acid dissociation constant pK _a was found to be 9.6. A clear isosbestic point could be obtained after purification at a unique pH for different concentrations. In contrast, in the absorbance measurements of acridine orange dodecyl, no isosbestic point was observed; this was attributed to the free long dodecyl chain. The overlap in the absorbance spectrum and the normalized fluorescence spectrum was found in many solvents around 500 nm. Neutral species were more likely to transfer into organic phase upon shaking. For fluorescence measurements both steady-state and time-resolved spectra were investigated. Results showed that at the interface a bathochromic shift happened to the maxima. This means that chromophore species aggregated at the interface as the dimer or trimer or the macro oligomer. Fluorescence polarization in bulk was calculated and found to be ca. 0.4. For interfacial measurements a dynamic volume method was applied to obtain the interfacial tension. An extrapolated Gibbs plot was obtained. Increasing bulk concentration increased the concentration at the interface to a finite value representing saturation.     

Photophysical and laser characteristics of pyrromethene 567 dye: Experimental and theoretical studies

Narrow-band laser performance of alcohol solutions of pyrromethene 567 (PM567) and rhodamine 6G (RH6G) dye was investigated using a home-made GIG-configured dye laser, excited by the second-harmonic radiation (at 532 nm) of a pulsed Nd:YAG laser. Higher laser efficiency was observed with PM567 dye (∼23% peak) in comparison to the commonly used RH6G dye (16.5%), in spite of much lower fluorescence quantum efficiency of the PM567 (0.83) vis-à-vis RH6G (0.98) dye solutions in ethanol. First principle-based electronic structure calculations were performed on PM567 dye in the ground (S ₀) and excited states (S ₁) using density functional theory to elucidate the structure and photophysical properties of the dye.     

Transverse pseudo-nonlinear effects measured in solid-state laser materials using a sensitive time-resolved technique

The fluorescence of different types of planthopper (Hemiptera) and moth (Lepidoptera), which constitute important Chinese agricultural pests, was investigated both in situ in a laboratory setting and remotely using a fluorescence light detection and ranging (lidar) system operating at a range of about 50 m. The natural autofluorescence of different species, as well as the fluorescence from insects that had been dusted with fluorescent dye powder for identification were studied. Autofluorescence spectra of both moths and planthoppers show a maximum intensity peak around 450 nm. Bleaching upon long-time laser illumination was modest and did not affect the shape of the spectrum. A single dyed rice planthopper, a few mm in size, could be detected at 50 m distance by using the fluorescence lidar system. By employing various marking dyes, different types of agricultural pest could be determined. We suggest that lidar may be used in studies of migration and movement of pest insects, including studies of their behavior in the vicinity of pheromone traps and in pheromone-treated fields.     

Regionalization of Plasma Membrane-Bound Flavoproteins of Cerebellar Granule Neurons in Culture by Fluorescence Energy Transfer Imaging

Flavoproteins are components of plasma membrane redox chains, which have been suggested to play major roles in neuronal activity and survival. We found that the red/orange autofluorescence of mature primary cultures of cerebellar granule neurons (8–9 days in vitro) was largely quenched by millimolar concentrations of dithionite added to the extracellular medium, and pointed out that nearly 50% of this autofluorescence was due to plasma membrane-bound flavoproteins. We report in this work that the lipophilic neuronal plasma membrane markers N-(3-triethylammoniumpropyl)-4-(4-(4-(diethylamino)phenyl)butadienyl)-pyridinium dibromide (RH-414) and N-(3-triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl)pyridinium dibromide (FM4-64) can form fluorescence energy transfer donor–acceptor pairs with flavoproteins with calculated R ₀ values between 3.7 and 4.2 nm. The quantification of the efficiency of fluorescence energy transfer with different concentrations of acceptor dyes has been worked out with re-suspended neurons. Using quantitative images of the neurons in culture, acquired with a CCD camera attached to an epifluorescence microscope, regionalization of the plasma membrane-bound flavoproteins of cerebellar granule neurons has been achieved from the quenching by dithionite of the fluorescence of the acceptor dye. The results unraveled that plasma membrane-bound flavoproteins are largely enriched in interneuronal contact sites forming clusters of 0.5–1 μm diameter size, which appears largely regionalized in the neuron's cell body.     

基于荧光光谱技术和GA-BP神经网络的对羟基苯甲酸甲酯钠含量的测定

对羟基苯甲酸甲酯钠是一种使用广泛的防腐剂,食用过量将会危害身体健康,因此各国对对羟基苯甲酸甲酯钠的用量有严格规定。采用FS920荧光光谱仪分析了对羟基苯甲酸甲酯钠在水和橙汁溶液中的荧光光谱特性,发现水溶液的荧光特征峰位于λ_ex/λ_em=380/510 nm,橙汁溶液两个荧光特征峰,分别位于λ_ex/λ_em=440/530 nm和470/530 nm,最佳激发波长为440 nm。从实验结果可以看出两者的特征峰发生了明显的变化。经分析得出,对羟基苯甲酸甲酯钠橙汁溶液与对羟基苯甲酸甲酯钠水溶液相比,荧光特征峰发生变化是由橙汁的荧光特性干扰引起的。为了准确测定鲜橙汁中对羟基苯甲酸甲酯钠含量,根据对羟基苯甲酸甲酯钠橙汁溶液在激发波长λ_ex=440 nm时的相对荧光强度和对羟基苯甲酸甲酯钠含量的关系,基于GA-BP神经网络构建了橙汁中对羟基苯甲酸甲酯钠含量检测数学模型,当网络训练过程中误差精度达到10-3时,网络输出与期望的相关系数为0.996,预测样本的平均回收率为99.52%,平均相对标准偏差为0.86%,预测结果较为理想。结果证明,当浓度范围为0.02~1.0 g·L-1时,荧光光谱技术和GA-BP神经网络相结合的方法能够准确地测定鲜橙汁中对羟基苯甲酸甲酯钠的含量,此方法具有新颖简便性,同时有望应用于一般饮品中对羟基苯甲酸酯类钠盐含量的快速测定。