Prism-based Spectral Imaging of Four Species of Single-molecule Fluorophores by Using One Excitation Laser

A prism-based imaging system for simultaneously detecting four species of single-molecule (SM) fluorophores was developed. As for the detection method, four spectrally distinct species of BigDye fluorophores were bound to 50-nm-diameter gold nanoparticles (AuNPs) to form AuNP/BigDye complexes. Four species of complexes were randomly immobilized on different fused-silica slides. BigDyes were excited by an argon-ion-laser (excitation wavelengths: 488 and 514.5 nm) beam through total internal reflection on the slide surface. SM fluorescence emitted from a complex was spectrally dispersed through a prism to form an SM spot elongated in the spectral direction on a charge-coupled device. A scattered light spot generated by the AuNP of the same complex under 594-nm laser illumination was used as a wavelength reference, and the SM fluorescence spectrum was obtained from the pixel-intensity pattern of the elongated SM spot. Peak locations of fluorescence spectra of all the observed SM spots were obtained, and their histograms were distinctly separated according to species. SM spots can thus be classified as one of four species according to their peak locations. By statistically analyzing the histograms, the classification accuracy was estimated to be above 93.8 %. The number of pixels in the spectral direction required for classifying four species of SM fluorophores was estimated to be 10. As for the conventional system (which uses two excitation lasers), 15 pixels are required. Using BigDyes as the four fluorophores (which consist of donors linked to acceptors and can be excited by just an argon-ion laser) is the reason that such a small number of pixels was achieved. The developed system can thus detect 1.5 times more SM fluorophores per field of view; that is, its throughput is 1.5 times higher. The approach taken in this study, namely, using BigDye with a prism-type system, is effective for increasing the throughput of DNA microarray-chip analysis and SM real-time DNA sequencing.     

Ferritin as a photocatalyst and scaffold for gold nanoparticle synthesis

The ferrihydrite mineral core of ferritin is a semi-conductor capable of catalyzing oxidation/reduction reactions. This report shows that ferritin can photoreduce AuCl₄ ⁻ to form gold nanoparticles (AuNPs). An important goal was to identify innocent reaction conditions that prevented formation of AuNPs unless the sample was illuminated in the presence of ferritin. TRIS buffer satisfied this requirement and produced AuNPs with spherical morphology with diameters of 5.7 ± 1.6 nm and a surface plasmon resonance (SPR) peak at 530 nm. Size-exclusion chromatography of the AuNP–ferritin reaction mixture produced two fractions containing both ferritin and AuNPs. TEM analysis of the fraction close to where native ferritin normally elutes showed that AuNPs form inside ferritin. The other peak eluted at a volume indicating a particle size much larger than ferritin. TEM analysis revealed AuNPs adjacent to ferritin molecules suggesting that a dimeric ferritin–AuNP species forms. We propose that the ferritin protein shell acts as a nucleation site for AuNP formation leading to the AuNP-ferritin dimeric species. Ferrihydrite nanoparticles (~10 nm diameter) were unable to produce soluble AuNPs under identical conditions unless apo ferritin was present indicating that the ferritin protein shell was essential for stabilizing AuNPs in aqueous solution.     

基于跃迁偶极矩取向特征研究乙醚溶液中分子的各向异性度

利用偏振荧光光谱和偏振激发光谱研究了乙醚溶液中荧光分子跃迁偶极距的取向特征.实验结果表明,在垂直线偏振光照射下,乙醚溶液发射出峰值位于305 nm的荧光谱,对应的最佳激励光波长为256 nm.由偏振激发光谱分析得到荧光体的吸收跃迁偶极矩和发射跃迁偶极矩间夹角α的变化规律,揭示了荧光去偏振过程：在粘性溶液中的荧光分子具有一定的偶极取向,α随激发光波长发生变化,当激发波长接近最佳激发光波长时,吸收跃迁偶极矩和发射跃迁偶极矩间趋于平行,荧光的退偏效果较弱,偏振度最大.研究结果能为分子空间取向特征的理论研究提供参考.     

Formation and encapsulation of gold nanoparticles using a polymeric amine reducing agent

We report on the use of poly(allylamine) hydrochloride (PAH) as a reducing agent for the controlled formation of gold nanoparticles (AuNPs) in the size range of 5–50 nm. The formation of AuNPs using this polymer matrix allows for the AuNPs to be imbedded in the polymer matrix, once formed. The kinetics of AuNP formation are shown to be pseudo first-order in [HAuCl₄] at room temperature. The kinetics of AuNP formation are controlled by the ratio of reducing agent to HAuCl₄ as well as the overall concentration of the PAH and HAuCl₄. Additionally, at low PAH:HAuCl₄ mole ratios, the plasmon resonance wavelength can be controlled through the ratio of the reactants. This plamson resonance shift is shown to be related to AuNP size by means of TEM imaging data on the AuNPs.     

ICP-MS: a powerful technique for quantitative determination of gold nanoparticles without previous dissolving

A direct and simple inductively coupled plasma mass spectroscopy (ICP-MS) method for the determination of gold nanoparticles (AuNP) with different particle sizes ranging from 5 to 20 nm and suspended in aqueous solutions is described. The results show no significant difference compared to the determination of the same AuNPs after digestion, as claimed by the literature. The obtained limit of quantification of the method is 0.15 μg/L Au(III) that corresponds to 4.40 × 10⁹ AuNP/L, considering spherical AuNPs 15 nm sized. Spike recovery experiments have shown that the sample matrix is a significant factor influencing the accuracy of the measurement. Spike recoveries from 93% to 95% are found for AuNP samples prepared in trisodium citrate, while for deionized H₂O a spike recovery of around 80% was obtained. The sample preparation mode along with the ICP-MS parameters have been optimized and found to be crucial so as to achieve the required accuracy for the direct quantification of AuNP suspensions. The effect of the nanoparticle size upon the ICP-MS signal also was studied, and only significant differences due to the chemical environment and not to the AuNPs size were found.     

光纤面板窗多碱光电阴极荧光谱特性研究

论述了光致荧光的特点以及微光像增强器多碱光电阴极光致荧光的测量原理, 测量了光纤面板输入窗多碱光电阴极的荧光谱. 测试结果表明, 光纤面板窗的多碱阴极的荧光谱不是一条光滑的高斯型曲线, 而是在一条高斯型曲线上叠加了一些小的干涉峰的曲线. 原因是光纤面板窗所传输的荧光中, 有两束特殊的光线. 一束光为准直光, 另一束光为入射角刚好等于全反射临界角的反射光. 这两束光具有固定的相位差或光程. 当这两束光的相位差相差λ的整数倍时, 它们将干涉并产生干涉加强峰;当这两束光的相位差相差1/2λ的奇数倍时, 它们将干涉并产生干涉减弱峰. 如果在荧光谱的峰值波长处刚好产生干涉加强峰, 那么所测量的峰值荧光强度较其固有的峰值荧光强度要高; 反之, 如果在荧光谱的峰值波长处刚好出现干涉减弱峰, 那么所测量的荧光强度就小于其固有的荧光强度. 另外由于受到干涉的影响, 荧光曲线半峰宽也不能精确确定,所以在分析光纤面板窗光电阴极的荧光谱时, 要考虑到干涉因素的影响.     

Terbium(III) Chelate as an Efficient Donor for Multiple-Wavelength Fluorescent Acceptors

The emission spectra of the lanthanide chelates enable them to act as a donor for several acceptors emitting at different wavelengths. Fluorescence resonance energy transfer between terbium(III) chelate labeled antibody Fab fragment (donor) and a 17β-estradiol conjugated to Alexa Fluor 488, 555, 594 or 680 (acceptor) was employed to study the functionality of the terbium(III) chelate as an efficient donor for several acceptors emitting from green to far-red. During measurement, the sensitized emission of the acceptor was measured at acceptor specific wavelength. All the tested dyes proved to be efficient acceptors, and they were successfully used in the competitive homogeneous E2 assay. The highest signal to background ratio and the best assay performance was obtained with Alexa Fluor 680, due to the very low donor emission background at the far-red area. In addition, the sensitized emission of both Alexa Fluor 488 and 680 could be measured simultaneously without significant cross talk.     

Highly sensitive colorimetric and fluorescent sensor for cyanazine based on the inner filter effect of gold nanoparticles

Cyanazine residue poses a great threat to human health and its derivatives would remain in soils, natural waters, and other environmental domains for a long time. Herein, a simple, rapid, and ultra-sensitive analytical method for the determination of cyanazine (CZ) based on inner filter effect (IFE) of Au nanoparticles (AuNPs) on the fluorescence of CdTe quantum dots (QDs) is first described in this study. With the presence of citrate-stabilized AuNPs, the fluorescence of GSH-capped CdTe QDs was remarkably quenched by AuNPs via IFE. The fluorescence of the AuNP–CdTe QD system was recovered upon addition of CZ. CZ can adsorb on to the surface of AuNPs due to its cyano group that has good affinity with gold, which could induce the aggregation of AuNPs accompanying color change from red to blue. Thus, the IFE of AuNPs on CdTe QDs was weakened, and the fluorescence intensity of CdTe QDs was recovered accordingly. A good linear correlation for detection of CZ was exhibited from 0.05 to 9 μM, and the detection limit reached 0.1568 μM, which was much lower than the safety limit required by the USA, the UK, and China. In order to probe into the selectivity of AuNPs towards CZ over other pesticides, various frequently used pesticides were mixed with AuNPs. AuNP composite solution shows good selectivity towards CZ among other pesticides. This method was successfully carried out for the assessment of CZ in real samples with satisfactory results, which revealed many advantages such as high sensitivity, low cost, and non-time-consuming compared with traditional methods.     

Influence of excitation light wavelength on the photoluminescence properties for ZnO films prepared by magnetron sputtering

Highly orientated polycrystalline ZnO films were deposited on sapphire, silicon and quartz substrates at room temperature by r.f. magnetron sputtering. Different photoluminescence (PL) spectra were observed when excited with different wavelength light. A UV emission peak (356 nm) and a blue peak (446 nm) were generated for the films on sapphire, silicon and quartz substrates, and only the 446 nm blue emission appeared for the films on glass substrates when the wavelength of the excitation light was 270 nm. With increasing the wavelength of the excitation light up to 300 and 320 nm, the UV emission disappeared for films on various substrates and the wavelength of the PL peaks increased up to 488 and 516 nm, respectively. When the wavelength of the excitation light increased to 398 nm, the PL spectrum becomes a wide band that is consistent with three emission peaks.     

Raman spectroscopy of combinatory anticancer drug release from gold nanoparticles inside a single leukemia cell

Combinatory anticancer drug release from gold nanoparticles (AuNPs) in K562 human myeloid leukemia cells was performed using Raman spectroscopy. We fabricated the anticancer drug of imatinib as a BCR‐ABL tyrosine kinase inhibitor on AuNP surfaces along with a transferrin (Tf)‐targeting moiety to treat the leukemia cells. DNA topoisomerase I inhibitor topotecan was also assembled to monitor its fluorescence onto AuNPs. The linker group of 4‐carboxylic benzoic acid was used to conjugate to targeting the Tf protein. Our Raman data indicated that the drug molecules were not detached in the cell culture media but released after treatment with glutathione (2 mM). Intracellular distribution and release of the anticancer drug–AuNP conjugates in K562 cells were examined by both fluorescence microscopy and dark‐field microscopy with surface‐enhanced Raman scattering. Copyright © 2013 John Wiley & Sons, Ltd.     

CdS/PAMAM树形分子纳米复合材料的荧光性能研究

采用荧光发射法研究了不同Cd2+与树形分子的摩尔比(负载比)条件下硫化镉(CdS)/聚酰胺-胺(RAMAM)树形分子纳米复合材料(NCs)水溶液的荧光性能,并探索了其荧光产生的机理.结果表明:在波长330 nm光的激发下,CdS/G4.0-NH2PAMAM NCs的荧光发射光谱中出现了2个峰,一个是PAMAM树形分子的...     

Cr:LiSrAlF6 channel waveguides as broadband fluorescence sources

We report on the production and fluorescence of active channel waveguides in Cr:LiSrAlF₆. We have produced ∼10 μm wide and 5 μm high channel waveguides by He⁺ ion implantation, lithographic patterning and subsequent Ar⁺ ion sputtering. Diode-pumped waveguides emitted 13 μW of fluorescence light with a spectrum ranging from 760 nm to 900 nm at a pump power of 165 mW and a pump wavelength of 660 nm. The compact and cheap optical pump source is a main advantage of this fluorescence material. This makes Cr:LiSrAlF₆ channel waveguides a suitable candidate for a broadband fluorescence source in low-coherence interferometry and other applications in the near-infrared wavelength range. PACS 42.70.Hj; 42.79.Gn; 42.30.Wb     

Luminescent properties of semiconductor composite systems composed of erbium triphthalocyanine molecules and a silicon slot structure in the near-infrared region

Photoluminescence spectra of organic semiconductors based on mono-, bis-, and triphthalocyanine containing erbium as a complexing agent have been obtained in the range of 1–1.8 μm. Comparison of the spectral characteristics has shown that erbium triphthalocyanine has the highest photoluminescence quantum yield at a wavelength of 1.5 μm. To enhance this effect, composite materials based on erbium triphthalocyanine and a silicon slot structure have been synthesized, in which an additional increase in the photoluminescence signal near 1.14 μm has been observed. At the same time, no photoluminescence signal has been observed near the wavelength of 1.5 μm. This can be explained by taking into account the interaction of the erbium triphthalocyanine molecules with the adsorption centers of the silicon matrix.     

Visible photoluminescence of co-sputtered Ge–Si duplex nanocrystals

The photoluminescence (PL) characteristics of co-sputtered Ge–Si duplex nanocrystal films were examined under excitation by a 325-nm HeCd laser, combined with Raman and Fourier-transform infrared reflection spectra analysis. A broad visible PL spectrum from the as-deposited Ge–Si nanocrystal films was observed in the wavelength range 350–700 nm. Basically, the PL spectrum can be considered to consist of two distinct parts originating from different emission mechanisms: (i) the spectrum in the range 350–520 nm, consisting of characteristic double peaks at 410 and 440 nm with PL intensities decreasing after vacuum annealing, probably due to vacancy defects in Si nanocrystals; and (ii) the spectrum in the range 520–700 nm, consisting of a characteristic peak at 550 nm with a PL intensity not affected by vacuum annealing, probably due to Ge-related interfacial defects. No size dependence of PL peak energy expected from quantum confinement effects was observed in the wavelength range investigated. However, with an increase of crystal size, the PL peak intensity in the blue zone decreased. The PL intensity is found to be strongly affected by silicon concentration. A film heated in air has a different PL mechanism from the as-deposited and vacuum-annealed films. PACS 78.67.Bf; 81.05.Cy; 81.15.Cd     

Agglomeration behavior of lipid-capped gold nanoparticles

The current investigation deciphers aggregation pattern of gold nanoparticles (AuNPs) and lipid-treated AuNPs when subjected to aqueous sodium chloride solution with increasing ionic strengths (100–400 nM). AuNPs were synthesized using 0.29 mM chloroauric acid and by varying the concentrations of trisodium citrate (AuNP1 1.55 mM, AuNP2 3.1 mM) and silver nitrate (AuNP3 5.3 μM, AuNP4 10.6 μM) with characteristic LSPR peaks in the range of 525–533 nm. TEM analysis revealed AuNPs to be predominantly faceted nanocrystals with the average size of AuNP1 to be 35?±?5 nm, AuNP2 15?±?5 nm, AuNP3 30?±?5 nm, and AuNP4 30?±?5 nm and the zeta-average for AuNPs were calculated to be 31.23, 63.80, 26.08, and 28 nm respectively. Induced aggregation was observed within 10 s in all synthesized AuNPs while lipid-treated AuNP2 (AuNP2-L) was found to withstand ionic interferences at all concentration levels. However, lipid-treated AuNPs synthesized using silver nitrate and 1.55 mM trisodium citrate (AuNP3, AuNP4) showed much lower stability. The zeta potential values of lipid-treated AuNPs (AuNP1-L-1x/200, ??17.93?±?1.02 mV; AuNP2-L-1x/200, ??21.63?±?0.70; AuNP3-L-1x/200, ??14.54?±?0.90; AuNP3-L-1x/200 ??13.77?±?0.83) justified these observations. To summarize, AuNP1 and AuNP2 treated with lipid mixture 1 equals or above 1x/200 or 1x/1000 respectively showed strong resistance against ionic interferences (up to 400 mM NaCl). Use of lipid mixture 1 for obtaining highly stable AuNPs also provided functional arms of various lengths which can be used for covalent coupling.

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Graphical abstract Agglomeration behavior of gold nanoparticles before and after lipid capping

     

用光致荧光研究多碱阴极光电发射机理

本文介绍了多碱光电阴极的特点及其在微光像增强器中的应用,叙述了光致荧光的原理,探索了利用光致荧光方法来研究多碱阴极Na₂KSb膜层电子跃迁几率的方法,并测量了两个不同灵敏度多碱阴极的荧光谱及同一个多碱阴极在工作和非工作两种状态下的荧光谱.测试结果表明,多碱阴极的荧光强度与其电子跃迁的几率及阴极灵敏度成正比,同时多碱阴极在工作状态下,荧光强度比非工作状态下有所降低,原因是一部分跃迁电子逸出多碱阴极产生光电发射,而这部分电子不再回到基态,因此不再发出荧光.另外本文还测量了多碱阴极在不同波长激光激发条件下的荧光谱.结果表明,长波激发与短波激发相比,长波激发所获得的荧光强度更高,这说明长波激发产生跃迁电子的几率高,同时荧光谱峰值波长与激光波长的偏移较小,因此跃迁电子数多且能量损失小,有利于光电发射.将多碱阴极的荧光谱与多碱阴极的量子效率相比较,看出跃迁电子数量和所处能级这两个对光电发射过程有影响的关键因素中,能级因素对光电发射过程的影响更大.但对多碱阴极而言,由于短波激发时的电子跃迁几率低于长波激发时的电子跃迁几率,跃迁电子扩散过程中的能量损失较大,因此短波的量子效率随波长的增加而增加.实践证明,光致荧光是研究多碱阴极光电发射过程的一种有效手段,通过对多碱阴极荧光谱的研究,进一步揭示了多碱阴极的光电发射的机理,为进一步改进工艺和提高多碱阴极的灵敏度提供了重要的参考价值.     

Temperature–Light Dual‐Responsive Au@PNIPAm Core‐Shell Microgel‐Based Optical Devices

Au nanoparticle (AuNP) core particles coated with a poly(N‐isopropylacrylamide) (pNIPAm) shell (Au@pNIPAm) are synthesized by seed mediated free radical polymerization. Subsequently, a temperature–light‐responsive photonic device is fabricated by sandwiching the Au@pNIPAm particles between two thin layers of Au. The optical device exhibits visual color and characteristic multipeak reflectance spectra, where peak position is primarily determined by the distance between two Au layers. Dual responsivities of the photonic device are achieved by combining the photothermal effect of AuNPs core (localized surface plasmon resonance (LSPR) effect) and the temperature responsivity of the pNIPAm shell. That is, the pNIPAm shell collapses as the temperature is increased above pNIPAm's lower critical solution temperature, either by direct heat input or heat generated by AuNPs' LSPR effect. To investigate the effect of AuNPs distribution in the microgels on the devices' photothermal responsivity, the Au@pNIPAm microgel‐based etalon devices are compared with that fabricated by AuNP‐doped pNIPAm‐based microgels; in terms of response kinetics and optical spectrum homogeneity. The uniform Au@pNIPAm microgel‐based devices show a fast response and exhibit a comparatively homogeneous spectrum over the whole slide. These materials can potentially find use in drug delivery systems, active optics, and soft robotics.     

Chemical solution-process method to synthesize ZnO nanorods with ultra-thin pinheads and ultra-thin nanobelts

ZnO nanorods with 30 nm-diameter ultra-thin pinheads and ultra-thin nanobelts were successfully synthesized using a thiourea solution to etch nanorods and nancombs, which were obtained by a conventional thermal evaporation method. The materials obtained were investigated by field emission scanning electron microscopy and energy-dispersive X-ray fluorescence. The data shows that hydrogen ions play an important role in synthesizing ZnO nanorods with ultra-thin pinheads and ultra-thin nanobelts. Field emission plots indicated that the turn-on field was reduced from 2.10 V/μm to 1.55 V/μm after thiourea solution treatment at a current density of 0.1 μA/cm². Room-temperature photoluminescence spectra from ZnO nanostructures showed the PL spectrum peaks shifted towards short wavelengths with a large enhancement of UV bands compared with those of ZnO nanorods and nanocombs. PACS 75.55.Gs; 61.46.-w; 81.40.Wx; 78.55.-m; 78.60.Fi     

Molecular probe FET: Long-wavelength fluorescence excitation effect and proton transfer reaction

We studied the steady-state fluorescence spectra of solutions of FET (4′-(diethylamino)-3-hydroxyflavone) in acetonitrile that were excited at different temperatures by quanta with different energies located in the range of the main absorption band and in its long-wavelength wing. We found that, at room temperature, the emission intensity ratio of the bands of the normal and tautomeric forms, which are located at 505 and 570 nm, respectively, depends on the excitation wavelength. In the range of the main absorption band 300–360 nm, this ratio remains nearly the same, i.e., 1.45, while, upon excitation in the range of the long-wavelength wing 360–380 nm of the main band, it decreases to 1.33 at a wavelength of 460 nm. In this same range, a long-wavelength excitation effect that is unusual for liquid inviscid solvents at room temperature, i.e., a bathochromic shift of the entire short-wavelength emission band by 11 nm, manifests itself. We propose to explain these dependences using energy diagrams, which take into account the dependence of free energy on the orientational polarization of the polar solvent. The observed effect of the long-wavelength shift of the fluorescence spectrum with increasing excitation wavelength is explained in terms of the inhomogeneous broadening of electronic spectra of polar solutions, and it should be described using the scheme of energy states that takes into account sublevels of orientational broadening due to orientational dipole-dipole interactions of the fluorophore with nearest molecules of the polar solvent, as well as the relation between the fluorophore lifetime in the excited state and the dielectric relaxation time of solvent molecules in the field of the fluorophore dipole.     

Nucleobase-Specific Enhancement of Cy3 Fluorescence

We report on the role of dye–nucleobase interactions on the photophysical properties of the indocarbocyanine Cy3. The fluorescence efficiency and lifetime of Cy3 increase in the presence of all four nucleoside monophosphates. This behavior correlates with an increase in the activation energy for photoisomerization and a ∼4 nm red shift in the fluorescence spectrum. Changes are more dramatic for the purines (dAMP, dGMP) than the pyrimidines (dCMP, dTMP), and for the nonsulfonated cyanine (DiIC₂(3)) than the sulfonated dye (Cy3–SE). These results are consistent with a model in which Cy3–nucleoside π–π interactions decrease the efficiency of photoisomerization, increasing the efficiency of fluorescence. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.