Enhanced Fluorescence from Periodic Arrays of Silver Nanoparticles

Electron beam lithography was used to fabricate silver nanoparticle arrays and study the effects of geometrical properties of particles on metal-enhanced fluorescence. Nanoparticle size, shape, interparticle spacing, and nominal thickness were varied in a combinatorial pattern for investigation of the particle plasmon resonance effect on enhancement of fluorescence from three different fluorophores; Fluorescein, Cy3, and Cy5. A specific geometric property for optimal enhancement from each fluorophore was determined. For interparticle spacings greater or equal to 270 nm, the enhancement matched what is expected for a single-particle model. For those particles smaller than 210 nm, the enhancement was lower than for the larger spacing in the range studied. Triangular-shaped particles gave similar enhancement to those of square-shaped particles. This combinatorial pattern by e-beam lithography was found to be useful for studying how individual parameters enhance the fluorescence that are important for rational design of enhanced fluorescence sensors.     

碲化物发光玻璃中银纳米颗粒表面等离激元增强铒离子发光的研究

有趣的贵金属表面等离激元的光学性质,尤其是在发光增强领域的表现,使得它已经成为全球的一个研究热点。表面等离激元就是光与贵金属中的自由电子相互作用时,自由电子和光波电磁场由于共振频率相同而形成的一种集体振荡态。该文研究了碲化物玻璃中银纳米颗粒的表面等离激元共振增强铒离子的发光。我们测量了吸收谱、激发谱、发光谱以及荧光寿命。首先,我们挑选365.5和379.0 nm吸收峰作为激发波长测量了385～780 nm波长范围的可见发光光谱,发现有4个发光峰,依次位于408.0,525.0,546.0和658.5 nm,容易指认出它们依次为铒离子的2H_9/2→4I_15/2,2H_11/2→4I_15/2,4S_3/2→4I_15/2和4F_9/2→4I_15/2的荧光跃迁;可以计算出[80 nm平均粒径纳米银的Er3+(0.5%)Ag(0.2%): 碲化物玻璃的样品A]的上述4个可见发光的峰值强度是[Er3+(0.5%): 碲化物玻璃的样品C]的大约1.44～2.52倍。同时,[50 nm平均粒径纳米银的Er3+(0.5%)Ag(0.2%): 碲化物玻璃的样品B]的上述4个可见发光的峰值强度是样品C的大约1.08～1.55倍。随后,我们挑选365.5和379.0 nm 吸收峰作为激发波长测量了928～1 680 nm波长范围的近红外发光光谱,发现近红外波段有两个发光峰,位于979.0和1 530.0 nm,容易指认出它们依次为铒离子的4I_11/2→4I_15/2和4I_13/2→4I_15/2的荧光跃迁;同样可以计算出样品A的上述2个近红外发光的峰值强度是样品C的大约1.43～2.14倍。同时,样品B的上述2个近红外发光的峰值强度是样品C的大约1.28～1.82倍。因此,发光的最大增强大约是2.52倍。从荧光寿命动力学实验,我们发现样品A的荧光寿命为τ_A(550)=43.5 μs,样品B的荧光寿命为τ_B(550)=43.2 μs,样品C的荧光寿命为τ_C(550)=48.6 μs。这些实验结果证实了τ_A≈τ_B ＜τ_C。它意味着样品(B)相对于样品(C)的发光增强是源于自发辐射增强效应。然而,它也意味着样品(A)相对于样品(B)的发光增强是源于纳米银颗粒的粒径尺寸r效应。也就是说当粒径尺寸r增大的时候,散射截面C_s和r6成正比,而吸收截面C_a和r3成正比,因此散射截面C_s增大的速度会远大于吸收截面C_a增大的速度,而散射截面C_s是荧光增强的原因,吸收截面C_a是荧光减弱的原因,所以随着银纳米颗粒尺寸的增大,其散射截面占主要部分,当发光材料和金属表面等离子体SP发生耦合时,能量快速的转移到金属表面等离子体SP上,而后被散射到远场,这有利于增强荧光。其综合的结果就导致了发光强度会随r的增大而增强。上述实验的结果对太阳能电池的光伏发电和生物物理应用等领域都有着很好的应用前景。     

New Insights in the Interpretation of Tryptophan Fluorescence

Origin of tryptophan fluorescence is still up to these days a quiz which is not completely solved. Fluorescence emission properties of tryptophan within proteins are in general considered as the result of fluorophore interaction within its environment. For example, a low fluorescence quantum yield is supposed to be the consequence of an important fluorophore–environment interaction. However, are we sure that the fluorophore has been excited upon light absorption? What if fluorophore excitation did not occur as the result of internal conformation specific to the fluorophore environment? Are we sure that all absorbed energy is used for the excitation process? Fluorescence lifetimes of Trp residues are considered to originate from rotamers or conformers resulting from the rotation of the indole ring within the peptide bonds. However, how can we explain the fact that in most of the proteins, the two lifetimes 0.5 and 3 ns, attributed to the conformers, are also observed for free tryptophan in solution? The present work, performed on free tryptophan and tyrosine in solution and on different proteins, shows that absorption and excitation spectra overlap but their intensities at the different excitation wavelengths are not necessarily equal. Also, we found that fluorescence emission intensities recorded at different excitation wavelengths depend on the intensities at these excitation wavelengths and not on the optical densities. Thus, excitation is not equal to absorption. In our interpretation of the data, we consider that absorbed photons are not necessary used only for the excitation, part of them are used to reorganize fluorophore molecules in a new state (excited structure) and another part is used for the excitation process. A new parameter that characterizes the ratio of the number of emitted photons over the real number of photons used to excite the fluorophore can be defined. We call this parameter, the emission to excitation ratio. Since our results were observed for fluorophores free in solution and present within proteins, structural reorganization does not depend on the protein backbone. Thus, fluorescence lifetimes (0.5 and 3 ns) observed for tryptophan molecules result from the new structures obtained in the excited state. Our theory allows opening a new way in the understanding of the origin of protein fluorescence and fluorescence of aromatic amino acids.     

Nuclear Excitation by X-Ray Emission of Femtosecond Laser Plasma

The proposed model of laser plasma emission spectrum formation enables us to determine the absolute value of the laser pulse to plasma emitted radiation conversion factor, profile of the emission spectrum, and frequency distribution of the intensity and energy in the emitted spectra. This is of interest for laser plasma diagnostics and provides a means for direct calculation of the number of excited nuclei in dependent on the parameters of laser pulse. This revised version was published online in August 2006 with corrections to the Cover Date.     

Nonresonant Nonlinearity of Silver Nanoparticles

Results of experimental investigations of the process of nonlinear refraction in composite materials based on silver nanoparticles in the region far from the plasma surface resonance of the nanoparticles (λ = 1064 nm) are presented. The reflection Z-scanning technique was used for measurements, which made it possible to determine both the sign and the values of nonlinear refractive indices and real parts of the 3rd-order nonlinear susceptibility of composite materials. The mechanism responsible for the process of nonlinear refraction in specimens is discussed. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 72, No. 5, pp. 650–653, September–October, 2005.     

Fluorescence Modulation of Acridine and Coumarin Dyes by Silver Nanoparticles

Silver nanoparticles were synthesized by chemical reduction of silver ions by sodium borohydride in the presence of poly-(N)-vinyl-2-pyrrolidone in solution of short chain alcohols. The nanoparticles are stable in 2-propanol, and the average diameter of the Ag colloid obtained in this solvent is about 6 nm. The photophysical properties of acridinium and coumarin dyes in 2-propanol are affected by the presence of silver nanoparticles. The interaction of silver nanoparticles with acridinium derivative leads to a spectral change of its intramolecular charge transfer (ICT) absorption band. The dye emission increases suddenly with the initial addition of the Ag metal nanoparticles, but at a high concentration of the colloid, static fluorescence quenching occurs with a progressive decrease of the fluorescence efficiency. Amino coumarin fluorescence is only quenched by the silver nanoparticles in solution.     

饱和激发下叶绿素荧光非线性变化及影响分析

叶绿素浓度是海洋初级生产力的重要指标之一,激光诱导荧光技术可以实现海水叶绿素浓度的快速测量。测量叶绿素浓度的传统激光诱导荧光原理,是利用叶绿素荧光与水体Raman散射的强度比值（I_F/R）进行反演,即叶绿素浓度n_chl＝CI_F/R,其中C为系统常量。这是依据叶绿素荧光685 nm、水体Raman散射强度都与激发光强呈线性关系。然而,该理论并没有考虑诱导荧光饱和现象的存在。当诱导激光强度达到一定程度后,685 nm荧光强度随激发光强非线性变化。另外,值得注意的是,水体Raman散射并不存在信号饱和现象。为了探讨饱和激发造成荧光非线性变化的影响,在激光诱导荧光技术测量叶绿素浓度的实验中,设计两种测量方案,即：不同激光功率诱导单一浓度样本的荧光测量,和固定激光功率时不同浓度样本的荧光测量。实验中利用Nd∶YAG三倍频激光355 nm激发获得叶绿素溶液的404 nm处 Raman散射和685 nm荧光。实验结果分为2部分进行讨论：（1）为了分析饱和激发造成荧光变化的非线性特性,通过调节激发光功率测量溶液的受激发射光谱,发现水体Raman散射强度与激发光强呈线性关系,而685 nm荧光强度出现饱和激发下的非线性变化。而且,随叶绿素浓度的增加,685 nm荧光的非线性趋势更为明显,Raman散射强度与激发光强的线性关系中斜率变小。数据分析表明,685 nm荧光数据拟合的4阶多项式和Raman散射效率值,可以定性地表征685 nm荧光的饱和程度。（2）考虑实际海洋激光雷达探测叶绿素浓度应用中存在饱和激发荧光非线性现象,为了分析荧光非线性对传统叶绿素浓度反演理论适用性的影响,在固定激发光强情况下对不同浓度叶绿素溶液的发射光谱进行测量。将激发光功率调节至52.00,80.70,132.10和197.30 mW·cm-2,获取相应激发光强下685 nm荧光与水体Raman散射的强度比值和叶绿素浓度之间的关系。实验表明,激发光强不变的情况下,685 nm荧光与水体Raman散射的强度比值,与叶绿素浓度仍满足线性关系。但是,在较高光强激发时,饱和激发造成的叶绿素荧光非线性变化,导致利用传统激光诱导荧光理论反演的叶绿素浓度值偏小。因此,需要对饱和激发下荧光非线性的影响进行修正,其关系为I_F/R=n_chl/C+C_F,修正值C_F不可忽略。另外,值得一提的是,修正关系中系统常量C随激发光强增加而增大。研究表明,饱和激发造成的荧光非线性,会对激光诱导荧光技术测量叶绿素浓度产生影响,但由于造成荧光非线性因素的复杂性,仅通过荧光数据拟合获得的多项式,无法定量说明其影响权重。然而,当激发光强不变时,可以实验测量获得基于激光诱导荧光原理的修正关系,从而准确反演叶绿素浓度。     

Quenched Emission of Fluorescence by Ligand Functionalized Gold Nanoparticles

A fluorescence-based detection scheme that uses ligand functionalized gold nanoparticles is proposed. The transduction scheme is based on the strong quenching of the fluorescence emission exerted by metallic surfaces on fluorophores positioned in their immediate vicinity (< 5 nm). Binding of fluorophore-labeled anti-biotin to biotinylated gold nanoparticles resulted in decreased fluorescence emission intensity. Subsequent competitive dissociation of labeled anti-biotin with D-biotin resulted in increased fluorescence emission intensity. These interactions occurred by means of specific molecular recognition because when the binding sites of anti-biotin were saturated with D-biotin prior to interaction with the gold nanoparticles; changes in the fluorescence emission intensity were not observed.     

ZnO纳米颗粒中的表面态发射特性研究

研究了不同粒径ZnO纳米颗粒样品(17～300nm)的时间光谱,通过对各粒径样品时间积分光谱的谱带结构进行高斯拟合解迭,发现光子能量位于ZnO谱带低能侧的高斯拟合成份Xc3的荧光中心波长随粒径的减小而红移,同时发光带的寿命也随之缩短.基于ZnO谱带低能侧的高斯拟合峰发光带强烈依赖于ZnO样品粒径的谱带特性,提出了与ZnO禁带内的表面态能级有关,同时研究表明,表面态在尺寸降到一定程度的纳米体系中起着重要的作用.     

Multi-Photon Fluorescence Correlation Spectroscopy: a Quantification of Tryptophan Methylester Solutions by Visible Emission

A multi-photon excitation fluorescence correlation system has been developed. The emission from tryptophan methylester solution was observed by this system and analyzed by the intensity correlation function of the visible emission, which originates from the two-photon excitation of photo products generated through a five-photon process. The intensity and the product concentration were proportional to the concentration of tryptophan methylester at a lower concentration range and thus the generation process is a single molecular reaction. The correlation analysis determined the concentration of tryptophan methylester down to 5 μM. The photo product generation from tryptophan solution was enhanced by a potassium iodide addition. These results suggest a new quantification method of tryptophan derivatives.     

银镜法制备具有表面增强红外活性Ag纳米粒子的探讨

利用银镜法在玻璃基底上沉积金属银制得表面增强红外材料,扫描电镜显示,沉积在基片上的银为球状颗粒,且平均直径在100nm左右,银纳米粒子形成一个个不相连的金属岛。以邻硝基苯胺为探针分子,用衰减全反射法(ATR)考察了邻硝基苯胺在银镜基底上的表面增强红外光谱。结果表明:邻硝基苯胺的最小检出质量约为40ng,增强因子约为70。制备银膜的最佳条件为:A gNO3质量分数为1%,反应温度为60℃,且加入表面活性剂可使银纳米粒子分散均匀。     

基于未修饰银纳米粒子光度法测定卡托普利

研究建立了一种简单、快速、高灵敏测定药物卡托普利的新方法。该方法基于卡托普利诱导未修饰的银纳米粒子的快速团聚,从而使银纳米溶胶从亮黄色变为红色,在395 nm处的吸光度呈线性降低。探讨了银纳米粒子的团聚机理。在优化条件下,该方法测定卡托普利的线性范围为1～35 μg·mL-1,检出限为0.7 μg·mL-1。应用于卡托普利药片含量的测定,获得了满意结果。     

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.     

二阶导数荧光分光光度法同时测定色氨酸和酷氨酸

本文描述了用二阶导数荧光光度法同时测定色氨酸和酪氨酸。在 p H 7.4的条件下 ,用 2 2 1nm作为激发波长 ,记录色氨酸和酪氨酸的发射光谱 ,并进行二阶导数处理。色氨酸在 318nm处 ,酪氨酸在 2 83nm处 ,二阶导数峰高与浓度成线性关系。色氨酸工作曲线的线性回归方程为 c =0 .0 0 0 7H - 0 .0 0 4,r =0 .996 4,线性范围为 0 .0 0 4到 0 .2 0 0μg . m L- 1 。酪氨酸工作曲线的线性回归方程为 c =0 .0 0 12 H -0 .0 0 40 ,r=0 .9971。线性范围为 0 .0 0 2到 0 .2 5 0 μg· m L- 1 。实验了 p H、温度和干扰离子对测定的影响 ,测定了苹果中的色氨酸和酪氨酸的含量 ,回收率分别为 (92 .0～ 10 4.0 ) %和 (98.70～ 10 2 .0 ) % ,相对标准偏差分别为 3.5 %和 2 .8%。     

Variable Frequency Microwave Synthesis of Silver Nanoparticles

Synthesis of silver nanoparticles based on a polyol process and variable frequency microwave (VFM) was investigated. Comparing to a thermal method, the reaction by VFM radiation was much faster. The effects of silver nitrate concentration, poly(N-vinylpyrrolidone) (PVP) concentration, reaction time and reaction temperature were studied. It was found that the higher concentration of silver nitrate, longer reaction time and higher temperature increased the particle size while the higher concentration of PVP decreased the particle size.     

分光光度法测定豆类及其粗蛋白质中的色氨酸

建立一种快速，准确测定豆类试样及其粗蛋白质中色氨酸含量的分光光度法，讨论最佳试验条件及试剂用量，反应产物在４１０ｎｍ处具有最大吸收波长ε＝１．１０３×１０~４Ｌ／ｍｏｌ·ｃｍ。回收率９３．５０％－１１０．０％，相对标准偏差小于２．６１％。     

ZnO纳米颗粒受激发射的时间分辨特性

在飞秒脉冲激光激发下,观察到了均匀沉积法获得的ZnO纳米颗粒的受激辐射现象,并从频域和时域两方面研究了ZnO纳米多晶的室温激射特性。氧化锌纳米颗粒中出现激子-激子散射导致的激射阈值为7.2 GW·cm-2,激射模式类似于F-P谐振腔模式,时域谱则表现为寿命曲线中出现快速衰减成分。与荧光的时间衰减曲线不同,P带时间衰减具有对称结构,高斯拟合结果只有几个ps,接近条纹相机的时间分辨率极限。研究ZnO纳米颗粒的受激发射与激光特性对揭示ZnO晶体的内部结构和激子激发态的性质、激光产生的机理等有重要意义。     

原子吸收光谱间接测定色氨酸的研究

色氨酸(Tryptophan,Trp)在碱性条件下与新生成的ZnS悬浮液反应,生成可溶性碱式色氨酸锌配合物,再用塞曼原子吸收光谱测定溶液中的总锌浓度即可间接的得到色氨酸的含量.优化了测定的最佳条件,在pH 9.10的2%硼砂底液中于室温下,使色氨酸试液与新生成的ZnS悬浮液反应22min,离心分离后测定上层清液的FAAS响应.在所选定的条件下色氨酸测定的线性浓度范围为0-18.5mg·mL-1,特征浓度为0.219mg·mL-1,检出限为1.414mg·mL-1.该法用于样品中的色氨酸测定,其相对标准偏差RSD为4.12%,方法的回收率在70.1%-120.3%之间.并对测定机理进行了探讨.     

紫外分光光度法同时测定色氨酸和酪氨酸

以0.1mol/L NaOH为溶媒,18种组成蛋白氨基酸在280-310nm波长仅色氨酸和酪氨酸有吸收峰.利用这一特性,提出了色氨酸和酪氨酸混合体系的紫外光谱的测定方法,同时用偏最小二乘法(PLS)和多元线性回归法(MLR)等化学计量学方法对光谱数据直接进行解析,结果都较为满意.本法已用于复方氨基酸注射液的测定,结果也...     

纳米微粒SnO2非线性光学特性的Z—扫描技术研究

利用Ｚ－扫描技术，以调ＱＮｄ：ＹＡＧ激光器作光源，首镒在非共振区域研究了ＳｎＯ２纳米微粒的三阶非线性光学性质，获得了三阶非线性极经系数ｘ＾（３）与双光子吸收系数β等重要依据，并对其非线性响应机制进行了探讨。