Advances in Surface-Enhanced Fluorescence

We report recent achievements in metal-enhanced fluorescence from our laboratory. Several fluorophore systems have been studied on metal particle-coated surfaces and in colloid suspensions. In particular, we describe a distance dependent enhancement on silver island films (SIFs), release of self-quenching of fluorescence near silver particles, and the applications of fluorescence enhancement near metalized surfaces to bioassays. We discuss a number of methods for various shaped silver particle deposition on surfaces.     

一种定征固体板表面性质的兰姆波方法

基于Ritec-SNAP系统建立了兰姆波实验系统，分析了用兰姆波幅频特性定征固体板表面性质的可行性。实验结果表明，用高阶、多模兰姆波的幅频特性可准确定征固体板表面性质的变化。     

Treatment of roughness and concentration gradients in total Reflection X-ray fluorescence analysis of surfaces

Total Reflection X-ray fluorescence (TXRF) spectrometry, a new technique for surface and layer analysis, was originally confined to ideal smooth interfaces. In practice, however, one has to cope with more or less rough surfaces. Therefore, modelling calculations have been conducted to consider the consequences of residual roughness on the fluorescence signal at grazing incidence. The model used was verified experimentally on surfaces which exhibit peak-to-valley roughnesses ranging from 5 to 4000 nm. In addition, concentration changes occurring in the zone of roughness in the surface layer of a high grade steel after exposure to nitric acid were determined.     

Size-dependent interaction of gold nanoparticles with transport protein: A spectroscopic study

Gold nanoparticles of different sizes have been synthesized using sodium citrate as a reducing agent for tetrachloroauric (III) acid. The formed gold nanoparticles have been characterized by the UV-visible and transmission electron microscopy (TEM) measurements. The different sized gold nanoparticles have been used to study the interaction with model transport protein, bovine serum albumin (BSA). Experimental results reveal that BSA molecules adsorbed on the metallic surfaces, suffer strong quenching of their fluorescence and the rate of quenching efficiency is different for different particle size. The analysis of the quenching results has been performed in terms of the Stern-Volmer equation. The mechanism of quenching of fluorescence has been explained. The extent of adsorption of BSA on the gold nanoparticles has been estimated.     

Eu(TTFA)3掺杂SiO2胶体球的制备与荧光光谱特性

制得了稀土配合物Eu(TTFA)3掺杂的SiO2胶体球,并用透射电子显微镜（TEM）和荧光分光光度计对其形貌和荧光光谱特性进行了详细地研究.TEM形貌分析表明,掺杂SiO2胶体颗粒具有球状的外形和光滑的表面,平均直径约为300 nm.荧光光谱分析表明,该稀土配合物掺杂胶体颗粒具有Eu3+离子典型的荧光光谱特性,荧光寿命约为290 μs.进一步分析实验数据,发现在Eu(TTFA)3掺杂浓度为质量分数0.5～1.5的范围内,荧光强度随着掺杂浓度的增加而增加,而荧光寿命几乎不随掺杂浓度而变化.     

Fluorescent Profiling of Venom-Selected Cobra Species

The aim of this paper was to characterize and compare spitting cobras of Africa. The autofluorescence of Naja ashei and Naja nigricollis venoms was investigated by atomic force microscopy and fluorescence analysis. The synchronous fluorescence analysis showed higher endogenous fluorescence of Naja ashei venom in comparison with Naja nigricollis venom. The atomic force microscopy revealed different surfaces of compared venoms as a result of different amounts of proteins. Our results confirmed different structures of Naja nigricollis and Naja ashei venom. The practical advantages of selected techniques are high sensitivity, minimal quantity of venom, and capability to measure the spitting cobra by autofluorescence.     

Resolution enhancement of random adsorbed single-molecule localization based on surface plasmon resonance illumination

Single-molecule localization (SML) is a powerful tool to overcome the diffraction limit in optical imaging, because the fluorescence emitted by single molecules can be observed with nanometer accuracy when the optical background and associated noise are made sufficiently small. Random adsorbed SML has been successfully demonstrated for superresolution imaging on metal surfaces. To optimize the random adsorbed SML, we developed a new illumination method based on surface plasmon resonance (SPR). The enhancement of the fluorescence signal and the reduction of background noise were achieved simultaneously. A high localization resolution of 15?nm was demonstrated with this new SPR illumination system.     

Adsorption of ethylenediamine on clean and oxygen covered Fe/Ni(100) surfaces studied by XPS

The interaction of ethylenediamine with Fe/Ni(100) surfaces oxidized to various extents has been studied in the temperature range 260–450 K by means of X-ray photoelectron Spectroscopy. The use of ~ 1 monolayer of Fe enables us to characterize the oxidation states of the topmost layer atoms unambiguously, based on the XPS spectra using a conventional spectrometer. On clean and c(2 × 2)-O surfaces the ethylenediamine can dissociate the N-H bond at 260 K. On heating the adlayer to 340 K the dissociation was further developed. On the surfaces whose Fe atoms were oxidized to FeO/Ni(100) and further, only molecularly adsorbed species were present at 260 K and desorbed partly without dissociation of the N-H bond after heating to 340 K.     

Gilding and pigments of Renaissance marble of Abatellis Palace: non‐invasive investigation by XRF spectrometry

Most of the artworks constituting the collection of Renaissance statuary of Abatellis Palace in Palermo (Sicily) show evidence of colour layers and fragments of gold foil that probably once covered the whole marble surface. The restoration of some of these statues has allowed to carry out archaeometric studies about the painting technique and to highlight the original materials and inclusion present on the precious marbles by two famous Italian sculptors of the Renaissance, Francesco Laurana and Antonello Gagini. The measurements have been performed in situ through the integrated use of two non‐invasive techniques: visible fluorescence stimulated by ultraviolet light and X‐ray fluorescence. The ultraviolet‐induced fluorescence analysis has provided additional information on the conservation status of marble surfaces by differentiating the pictorial materials and highlighting the presence of gilding and pigment traces through their characteristic fluorescence response. The observation in ultraviolet light has been used as valuable guide for the identification of the significant points to be analysed by X‐ray fluorescence to characterise the original materials. X‐ray fluorescence measurements have cast light about their chemical composition and stratigraphical structure. Pictorial layers were identified: vermilion for red layers, blue pigment based on copper for blue layers and pure gold leaf for gilding layers. Principal component analysis of the data was capable of clustering the different painting materials, discriminating through their chemical content. The results represent an important scientific support both to the hypotheses about the original look of the artworks and to the resolution of restoration and conservation questions still open. Copyright © 2012 John Wiley & Sons, Ltd.     

Fluorescence Enhancements on Silver Colloid Coated Surfaces

We observed a strong, more than 16-fold, enhancement of Texas Red-labeled BSA fluorescence emission when deposited on silver colloid coated surfaces (SCCS). The same labeled protein deposited on silver island films (SIFs) showed an approximate 8-fold fluorescence enhancement. The lifetimes of Texas Red-BSA fluorescence are significantly shorter on silvered surfaces than on uncoated quartz substrate indicating a strong change in radiative decay rate of the dyes. We also observed a 36-fold increased brightness of overlabeled fluorescein-HSA deposited on silver colloid coated surface. Stronger enhancement observed for overlabeled Fl-HSA protein indicates that presence of silver particles partially decreased self-quenching. Our results indicate that surfaces coated with silver colloids are valuable substrates for metal-enhanced fluorescence.     

Parameters optimization for biological molecules patterning using 248-nm ultrafast lasers

Laser-induced forward transfer has been used for the deposition of photoactive biotin in micron-scale patterns. The process uses a 500 fs pulsed KrF laser beam to transfer small amounts of a liquid solution target as micron-size droplets to a substrate placed parallel and in close proximity to it. The biomolecules remain active after the transfer; this is demonstrated through fluorescence assays. In addition to the laser parameters, those regarding the target composition and the receiving surface for the miniaturization of the transferred patterns have been studied and optimized. Droplets as small as 5 μm have been obtained by reducing the target thickness and transfer energy; by increasing the percentage of glycerol added in the biomolecules solution and by using hydrophobic surfaces as receiving substrates. The influence of the glycerol addition and the hydrophobicity of the receiving surfaces on the activity of the transferred biomolecules have also been studied.     

Effect of surface roughness on ellipsometry of aluminum

Controlled surface roughness experiments have been performed on aluminum to determine the effect on ellipsometric parameters. To help characterize the surfaces SEM, light scattering, photo electron emission, surface potential difference and water contact angle measurements were made. Comparison of experimental results with recent theory is also presented.     

Methods for the Modification and Characterization of Oxide Surfaces

This paper presents a general overview of the chemical modificationof oxide surfaces with particular emphasis on the silanization/hydrosilationmethod. A number of materials have been synthesized that have hydrophobic,hydrophilic, chiral and liquid crystal surfaces. The most frequently usedoxide in these materials is silica with chromatography as the primaryapplication for the chemically modified oxide surfaces. Also presented is anoverview of a number of methods used to characterize modified oxide surfacessuch as elemental analysis, diffuse reflectance infrared Fourier transformspectroscopy, solid-state NMR, electron spectroscopy for chemical analysis,scanning electron microscopy and atomic force microscopy. The particularadvantages of each method for the characterization of modified oxidematerials are discussed.     

聚电解质包覆的铕(Ⅲ)配合物@SiO2荧光纳米粒的合成与性质

以二苯甲酰甲烷(DBM)、邻菲罗琳(phen)和丙烯酸(AA)为配体,制备了铕的配合物Eu(Ⅲ)(DBM)2-(phen)(AA).利用St(o)ber法合成了SiO2纳米粒.通过超声辅助,将脂溶性的强荧光铕配合物吸附到SiO2纳米粒上,再包覆阳离子聚电解质聚二烯丙基二甲基氯化铵(PDAC)和阴离子聚电解质聚丙烯酸(P...     

Study of plasma membrane heterogeneity using a phosphatidylcholine derivative of 1,6-diphenyl-1,3,5-hexatriene [2-(3-(diphenylhexatriene)propanoyl)-3-palmitoyl-l-α-phosphatidylcholine]

The fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH) and DPH derivatives have been used to characterize structural and physicochemical properties of specific membrane domains. Steady-state and fluorescence decay measurements of three probes, DPH (1,6-diphenyl-1,3,5-hexatriene), TMA-DPH [1-(4-trimethyl-ammonium-phenyl)-6-phenyl-1,3,5-hexatriene], and a phosphatidylcholine derivative of DPH, DPH-pPC [2-(3-(diphenylhexatriene)propanoyl)-3-pamitoyl-L--phosphatidyl choline], have been performed in erythrocyte membranes and in lymphocyte plasma membranes. The steady-state fluorescence polarization of the three probes showed a similar trend in both membranes. In fact either in erythrocyte or in lymphocyte plasma membranes the fluorescence polarization values of DPH-pPC and TMA-DPH were similar, but significantly higher with respect to DPH. A better characterization of erythrocyte and lymphocyte plasma membranes was possible by using fluorescence decay measurements. The data suggest the possible use of different DPH derivatives to characterize specific domains in biological membranes.     

Fluorescence properties of dyes adsorbed to silver islands,investigated by picosecond techniques

The fluorescence properties of dye molecules (rhodamine 6G and erythrosin) adsorbed on pure glass surfaces and on silver islands films are investigated by cw and picosecond time-resolved methods. On pure glass surfaces we observe concentration quenching below a critical intermolecular distance (reduction of the fluorescence power per molecule as well as shortened and non-exponential fluorescence decay). On silver islands films the shortening in fluorescence lifetime is more drastic and is nearly independent of the intermolecular distance. This behavior suggests an electrodynamic interaction between dye monomers and plasmons in the metal particles, modified by a damping influence of dye dimers.     

Synthesis of large-scale GaN nanowires by ammoniating Ga2O3 films on Co layer deposited on Si（111） substrates

A mass of GaN nanowires has been successfully synthesized on Si（111） substrates by magnetron sputtering through ammoniating Ga2O3/Co films at 950℃. X-ray diffraction, scanning electron microscopy, high resolution transmission electron microscope and Fourier transformed infrared spectra are used to characterize the samples. The results demonstrate that the nanowires are of single-crystal GaN with a hexagonal wurtzite structure and possess relatively smooth surfaces. The growth mechanism of GaN nanowires is also discussed.     

Metal-Enhanced Fluorescence from Plastic Substrates

We report the first findings of Metal-Enhanced Fluorescence (MEF) from modified plastic substrates. In the past several years our laboratories have reported the favorable effects of fluorophores in close proximity to silver nanoparticles. These effects include, enhanced fluorescence intensities, (increased detectability), and reduced lifetimes, (enhanced fluorophore photostability). All of these reports have featured silver nanostructures and fluorophores which have been immobilized onto clean glass or quartz surfaces. In this report we show how plastic surfaces can be modified to obtain surface functionality, which in turn allows for silver deposition and therefore metal-enhanced fluorescence of fluorophores positioned above the silver using a protein spacer. Our findings show that plastic substrates are ideal surfaces for metal-enhanced phenomena, producing similar enhancements as compared to clean glass surfaces. Subsequently, we speculate that plastic substrates for MEF will find common place, as compared to the more expensive and less versatile traditional silica based supports.     

Size and stability of liposomes: A possible role of hydration and osmotic forces

Dynamic light scattering and electrophoretic mobility measurements have been used to characterize the size, size distribution and zeta potentials (ζ-potentials) of egg yolk phosphatidylcholine (EYPC) liposomes in the presence of monovalent ions ( Na⁺ and K⁺). To study the stability of liposomes the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory has been extended by introducing the hydrated radius of the adsorbed ions onto the liposome surfaces. The decrease of liposome size is explained on the basis of the membrane impermeability to some ions which generate osmotic forces, which leads to evacuate water from liposome inside.     

Infrared spectroscopy of self-assembled monolayer films on silicon

Infrared vibrational spectroscopy in an attenuated total reflection (ATR) geometry has been employed to investigate the presence of organic thin layers on Si-wafer surfaces. The phenomena have been simulated to show there can be a field enhancement with the presented single-reflection ATR (SR-ATR) approach which is substantially larger than for conventional ATR or specular reflection. In SR-ATR, a discontinuity of the field normal to the film contributes a field enhancement in the lower index thin film causing a two order of magnitude increase in sensitivity. SR-ATR was employed to characterize a single monolayer of undecylenic acid self-assembled on Si(1 1 1) and to investigate a two monolayer system obtained by adding a monolayer of bovine serum albumin protein.