Fluorescence of the Protein of the Prolamellar Bodies of Etioplasts

In the high-purity membranes of the prolamellar bodies (PLB) of etioplasts, the main protein component of which is protochlorophyllide oxidoreductase (POR), protein fluorescence in the 320–350-nm region with a maximum in the excitation spectrum at 278–280 nm has been revealed. The microsurrounding of protein fluorophors in an acidic medium (pH 4.0) is the most hydrophobic. Transfer of energy from the protein to pyrene present in the lipid of the PLB membranes is observed, but fluorescence of protochlorophyllide (Pd) is absent when pyrene is excited via protein. A change in the content of Pd and carotenoids in the membranes of PLB influences the degree of excimerization of the acceptor molecules of pyrene (inverse dependence). The character of interaction of POR with a membrane with allowance for the data on its primary and secondary structures is discussed.     

Fluorescence of pyrene in galactolipid liposomes and native membranes of etioplasts

The fluorescence parameters of pyrene and pyrenedecanoic acid in lipid micelles and native membranes of etioplasts whose basic lipid components are mono- and digalactosyldiacylglycerol (MGDG and DGDG) have been investigated. The vibrational structure of the emission spectra of pyrene (λ_exc = 337 nm) in the 370–400-nm range points to the hydrophilic microsurrounding of pyrene molecules in all types of the investigated membranes. The absence of MGDG from liposomes leads to a decrease in the degree of excimerization of a probe. In native membranes, this parameter increases with increasing content of pigments. Pyrene covalently bound to decanoic acid is found in the membranes of prolamellar bodies and in artificial monolayer micellae in an even more hydrophilic microsurrounding than the molecules of a free probe, and a decrease in the degree of eximerization is observed thereby. The specific features of localization of the investigated probes in plastid membranes are discussed. Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 67, No. 3, pp. 344–349, May–June, 2000.     

Fluorescence Enhancement and Photostability of Novel Pentamethine Cyanines in Nafion-Na+ Membranes

Four novel pentamethine cyanines have been investigated for their photophysical behaviors and photochemical stability in Nafion-Na⁺ membranes. The incorporation processes of these dyes into Nafion membranes are diffusion-controlled and the apparent diffusion constants have been determined. Fluorescence quantum yields and fluorescence lifetimes of all tested dyes in Nafion membranes have been determined and significant fluorescence enhancements were observed with respect to their fluorescence in aqueous solution. Photolysis of these dyes suggested these dye molecules are stabilized significantly by Nafion membranes. Furthermore, the fluorescence enhancements and the photostabilization of these dyes are more significant in dry Nafion membranes than those in wet ones.     

Investigation of the Dynamic Properties of the Lipid Layer of Intraplastid Membranes by Lipophilic Fluorescent Probes

The dynamic properties of the lipid layer of intraplastid membranes have been studied by analyzing the stationary and kinetic spectral polarization characteristics of the fluorescence of lipophilic probes of pyrene and diphenylhexatriene (DPH). Based on the data of the decay of the fluorescence anisotropy of pyrene, the value of the microviscosity of lipids in the membranes of prolamellar bodies (PLB) and protilakoids of etioplasts has been calculated. The pyrene molecules built into the membranes of etioplasts have a relatively high rotational mobility (stationary anisotropy r _s < 0.1). The DPH molecules rotate with difficulty in etioplast membranes (r _s > 0.3). After photoreduction of protochlorophyllide (Pd) in vivo, the rotation of the pyrene and DPH molecules in the membranes of prolamellar bodies becomes easier and this leads to a decrease in r _s. Illumination raised the degree of excimerization of the pyrene immersed into lipids (_exc = 337 nm), and the microsurrounding of the molecules of the probe in lipids became more hydropholic (the relationship between the vibronic maxima at 373 and 387 nm decrease). The set of data obtained points to a decrease in the microviscosity of the lipid layer of the membranes of prolamellar bodies as a result of illumination of sproutings.     

异丙醇-水配合液的荧光偏振和时间分辨特性

研究了紫外光照射下异丙醇-水配合液的偏振荧光光谱,以及不同荧光峰处光子强度随时间的衰变过程,计算了偏振度并讨论了其偏振特性,测试了不同峰位对应的荧光寿命并分析了其荧光发射特性.结果表明,异丙醇-水配合液在紫外光激励下发射的荧光为具有确定分子取向的部分偏振光,偏振度和各向异性度分别为0.542和0.441.在波长为220...     

Sensitized Fluorescence of Silver Nanoparticles in the Presence of Pyrene

The fluorescence of pyrene adsorbed onto the surface of the cetyltrimethylammonium-coated silver nanoparticles was studied. Pyrene molecules adsorbed on freshly prepared silver particles were found to be in close proximity to silver surface thus providing the possibility of energy transfer from excited pyrene to silver cores of the particles. In that case along with the expected fluorescence of pyrene we observed the fluorescence of the silver nanoparticles induced by the excited pyrene molecules. In due course the restructuring of the cetyltrimethylammonium layer resulted in moving of pyrene molecules away from silver surface and simultaneous disappearance of the silver nanoparticles fluorescence band. These data strongly support the recent hypothesis of fluorophore-plasmon coupled emission.     

355 nm纳秒激光作用下荧蒽的时间分辨荧光光谱特征

多环芳烃由于具有三致(致癌、致畸、致突变)特性,其在环境中的检测受到人们广泛关注。利用时间分辨光谱技术,研究了荧蒽乙醇溶液的荧光光谱随延时时间和门宽改变的特性。研究了不同浓度荧蒽的时间分辨荧光光谱特性,以原始浓度的荧蒽为初始溶液,通过逐级稀释的方式,最终将原始溶液稀释16倍,拟合了不同稀释倍数下的荧蒽荧光强度衰减随延时时间变化的动力学曲线,得到了不同浓度荧蒽的拟合荧光寿命。研究结果表明,荧蒽的荧光光谱特性与光谱仪探测器延时时间和门宽宽度密切相关。固定延时时间,随着光谱仪门宽宽度的变化,荧蒽的荧光强度随着门宽的增大而逐渐增强。固定门宽,改变延时时间的过程中,荧蒽的荧光强度随延时时间呈现先增大,后减小的趋势。荧蒽的荧光强度随延时时间的衰减过程符合指数衰减过程,将荧蒽乙醇溶液进行逐级稀释,荧蒽荧光强度与延时时间的衰减进行指数拟合后,得到不同稀释倍数的荧蒽乙醇溶液的衰减动力学参数,随着稀释倍数的增大,拟合得到的荧蒽荧光寿命增大。多环芳烃时间分辨光谱特征的研究,可以为环境中多环芳烃的检测提供技术基础,由于不同荧光物质具有特征的荧光寿命,因此,可以利用多环芳烃与环境中其他荧光物质的不同荧光寿命特性,准确识别环境中的多环芳烃污染物。     

Evaluation of pyrene fluorescence quenching in biological membranes

The redistribution between two excited forms of pyrene accompanying a change in the intramembrane probe concentration yields a set of fluorescence spectra that intersect at a single common (isobestic) point. Infensities of monomer and excimer fluorescence (I_m and I_e, respectively) are related by a linear dependence: I_e+kI_m=I_s. The coefficient k depends on the membrane type and temperature. Fluorescence intensity at the isobestic point (I_i) along with the proportional quantity I_s reflect the total steady-state level of excited pyrene forms and do not depend on the excimerization degree (I_e/I_m ratio), and can be used when estimating the quenching degree of pyrene fluorescence in biological membranes. Institute of Photobiology, Academy of Sciences of Belarus, 27, F. Skorina St., Minsk, 220072, Belarus. Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 64, No. 2, pp. 204–208, March–April, 1997.     

8-羟基喹啉铝高分子复合物薄膜的瞬态荧光特性与机理

对 8 -羟基喹啉铝 (Alq3)高分子复合物薄膜的的瞬态荧光特性和机理进行了研究 ,发现复合物薄膜的荧光寿命比 Alq3样品的寿命明显缩短 ,荧光峰值波长紫移 ,高分子基质引起的淬灭过程可能是导致复合物薄膜寿命降低的主要原因 ,而 Alq3分子与基质分子间的互作用则引起荧光峰值波长的移动。     

同时测量荧光光纤温度和应变的实验研究

利用掺杂铒元素荧光光纤的试验样品实验测试了荧光寿命的温度特性，并得到荧光寿命在不同温度下的应变响应关系.试验表明，荧光寿命会随着温度的升高而降低；与温度相比荧光寿命随应变的变化很小.提出了一个潜在的同时测量应变和温度的传感器模型.     

Spectral Properties of Bacteriochlorophyll c in Organisms and in Model Systems

Polarized absorption and fluorescence spectra of bacteriochlorophyll c and green photosynthetic bacterium Prostheecochloris aestuarii cells and cell fragments embedded in stretched polymer film were measured. In pigment samples the artificial oligomers of bacteriochlorophyll c (with absorption about 750 nm) and other forms of this pigment and bacteriopheophytin (with absorption at 670 nm) were present. In bacteria samples, embedded in polymer, oligomers were in high degree disaggregated and as a result the absorption about 670 nm was observed. Previously for similar sets of samples the decay of fluorescence excited only at one wavelength was analyzed on three exponential components, but exact lifetime values of these components for various samples were different. The aim of present paper was to check if these differences occur because of various contributions to decay from three well defined forms or if they were related to the existence of several pigment forms with slightly different lifetimes. The global analysis of data obtained for various excitation and observation wavelengths of fluorescence were done. From this analysis it follows that the second situation occurs. For a model system containing artificial oligomers the largest component of decay has a ⁴ of about 0.183 ns or 0.136 ns depending on observation wavelength. For the bacteria sample, in which the emission at 680 nm is the superposition from various pigments, global analysis done for various excitation wavelengths shows also that the values differ depending on the regions of fluorescence observation. From polarized spectra, it follows that in the model system the pigments absorbing at 670 nm are randomly distributed whereas oligomers are highly oriented. In bacteria fragments absorbing at 670 nm pigment molecules can be divided into two groups: one oriented along the axis of film stretching and the second practically randomly distributed. In living organisms, under some conditions, small amount of 670 nm pigments can be present and can work as excitation energy traps or as antenna transferring the excitation. Present results show that the role of various pools of 670 nm absorbing pigments can be different because of their differing orientation.     

Fluorescence studies on the conformation of litorin in solution and in the presence of model membranes

The conformation of the nonapeptide hormone litorin has been studied in buffer and in the presence of lipids, using static and dynamic fluorescence. The results obtained show that, in buffer, the hormone probably exists in a collection of flexible conformers, slowly interconverting between them. The marked changes observed in fluorescence spectra and lifetimes upon addition of dimyristoylphosphatidylserine vesicles clearly show that the peptide interacts with lipids assuming lipid specific conformations. Interestingly, no significative spectroscopic changes are produced by exposure to dimirystoylphosphatidylcholine vesicles both in the gel and liquid-christalline phases, suggesting a requirement for negatively charged lipids during the process of hormone-membrane interaction.     

磷酸盐钕玻璃的荧光寿命和损耗对激光增益特性的影响

实验研究了目前使用的Ｎ２１２２和Ｎ３１２２掺钕磷酸盐玻璃中不同的ＯＨ基浓度对玻璃荧光寿命的影响，同时从实验和理论上研究了荧光寿命和损耗对激光增益特性的影响，结果表明，增加玻璃的荧光寿命是提高增益性能的一个重要方法，当玻璃的荧光寿命增加１０％时小信号增益系数提高５％左右，提高钕玻璃的荧光寿命，可适当降低对损耗的要求，也能够保持它的增益性能不变。     

Determination of the Constants of Binding of Acridine Dyes with Micelles of Sodium Dodecyl Sulfate Using the Quenching of Delayed Fluorescence by Heavy Atoms

The constants of binding dye molecules with the micelles of sodium dodecyl sulfate are determined using quenching of delayed fluorescence of acridine dyes by sodium iodide in aqueous–micellar solutions. Kinetic equations have been composed that describe the processes of deactivation of the excited states of dyes. By solving these equations at the concentration of the quencher sodium iodide corresponding to the minimum lifetime of triplet states and at the concentration of micelles corresponding to the least value of the delayed fluorescence quenching rate constants, we obtained the constants of binding dyes with micelles equal to 1.3·10⁷, 2.9·10⁷, and 3.1·10⁷ M^–1 for trypaflavine, acridine orange, and acridine yellow, respectively. We calculated the rate constants of quenching of the triplet states of the molecules of dyes by iodide ions (I ^–) that decreased in transition from trypaflavine to acridine orange and acridine yellow.     

TRANES Spectra of Fluorescence Probes in Lipid Bilayer Membranes: An Assessment of Population Heterogeneity and Dynamics

Time-resolved fluorescence of eight fluorescence probes were studied in EggPC bilayer membrane vesicles. Emission wavelength dependent fluorescence decays were analyzed in a model-independent way to obtain time resolved area normalized emission spectra (TRANES). The TRANES spectra of the probes studied were classified into four types: (1) spectra that are identical at all time (one emissive species), (2) spectra that show an isoemissive point (two emissive species), (3) spectra that shift continuously with time (slow solvation dynamics or multiple species), and (4) spectra that shift for a short time and thereafter one or two emissive species are indicated. The TRANES spectra of these eight probes, except RH421, belong to the type 1, 2, or 4. The continuous shift of the TRANES spectra that was observed for the probe RH421 is attributed to multiple ground state species and not due to slow solvation dynamics.     

Steady-State and Time-Resolved Spectroscopic Characteristics of Mesoionic Oxazolones Solutions

Three new mesoionic oxazolo[3,2-b]pyridazin-2-one derivatives, in different solutions have been investigated by UV-Vis absorption, steady-state and time-resolved fluorescence methods. The effect of substituents on the extension of conjugation of the pi-electrons from mesoionic oxazolone has been evidenced by bathochromic shifts of the absorption and fluorescence maxima positions. The fluorescence decay data could be fitted to single-exponential or double-exponential function. The lifetime values are much higher in aprotic polar solvents and in the case of the derivatives that present an extension of the conjugation of pi-electrons. The properties of the compounds present a solvent dependence, being tested in micellar solutions as potential molecular probe "sensitive" to the environment polarity.     

应用小波边缘检测技术分析荧光寿命显微像

研究了小波图象边缘检测技术在荧光寿命显微像分析中的应用，利用小波分析的多尺度特性进行了荧光寿命显微像的边缘提取，结果表明，这种方法非常有效。     

Monitoring Micelle Formation of Nonionic Polyurethane in Water by Fluorescence Spectrophotometry

A series of nonionic polyurethanes were dissolved in water and their miceller self-assembly behaviors were characterized by aid of a fluorescence spectrophotometer. Fluorescence and resonance light scattering techniques were carried out for characterizing the micelles, and information such as aggregation number, critical micelle concentration, micropolarity, and micelle volume change was obtained. The micropolarity decreased as the aggregation number increased. The nonionic polyurethane micelle volume with two long ethoxyl chains per polyurethane molecule was smaller than the one with short chains, even with the same aggregation number, and was less polar. The micropolarity depended on both the aggregation number and the volume of a single micelle.     

LB膜中聚集体的类型及其荧光特性研究

采用稳态和时间分辨芝光研究了不同类型的ＬＢ多层膜中分子聚集特性。在半花菁／花生酸交替膜中,花生酸层的隔离使得半花菁分子之间的相互作用主要发生在同一层中,形成Ｈ形成Ｈ聚集体,荧光光谱蓝移;在纯半花菁Ｙ型和Ｚ型膜中,较强的层间相互作用使分子形成Ｊ聚集体,导致荧光光谱发生显著红移。     

Inter- and Intramolecular Dynamics of Pyrenyl Lipids in Bilayer Membranes from Time-Resolved Fluorescence Spectroscopy

The analysis of time-resolved fluorescence of monopyrenyl phospholipid monomer and excimer emission is reviewed using a model based on the diffusion equation for bimolecular reactions in two-dimensional planar membrane bilayers. This aspect is illustrated by the analysis of a real experiment. The method can also be extended to short-range diffusion, which can be measured for dipyrenyl phospholipids in membrane bilayers. The latter aspect is illustrated by the analysis of a simulated experiment to show the experimental requirements to recover the desired parameters.