Optimized Synthesis of PEG-Encapsulated Gold Nanorods for Improved Stability and Its Application in OCT Imaging with Enhanced Contrast

Gold nanorods (GNRs) are synthesized with a surfactant template, which often poses toxicity issues for biomedical applications. In addition, blue shift of longitudinal surface plasmon resonance (LSPR) peak of GNR is an inherent problem that needs to be addressed for time-course studies. In this work, we resolve these issues by optimizing the encapsulation of GNRs with polyethylene glycol (PEG) where biocompatibility is improved by ～20 % and blue shift over a period of 8 days is reduced from 20 nm in the case of CTAB-GNR to 2 nm for PEG-encapsulated GNR. The encapsulated GNRs were then bioconjugated for targeted dark-field imaging of cancer cells. As an application, we also demonstrate the contrast-enhancing capability of GNRs in optical coherence tomography (OCT) imaging of tumor xenograft where the LSPR closely matches the OCT excitation wavelength. Our study proves that incorporating GNRs enhances the contrast of tumor tissue interfaces along with a considerable broadening in OCT depth profile by six times.     

Enhancing LSPR Sensitivity of Au Gratings through Graphene Coupling to Au Film

A particular interesting plasmonic system is that of metallic nanostructures interacting with metal films. As the localized surface plasmon resonance (LSPR) behavior of gold nanostructures (Au NPs) on the top of a gold thin film is exquisitely sensitive to the spacer distance of the film-Au NPs, we investigate in the present work the influence of a few-layered graphene spacer on the LSPR behavior of the NPs. The idea is to evidence the role of few-layered graphene as one of the thinnest possible spacer. We first show that the coupling to the Au film induces a strong lowering at around 507 nm and sharpening of the main LSPR of the Au NPs. Moreover, a blue shift in the main LSP resonance of about 13 nm is observed in the presence of a few-layered graphene spacer when compared to the case where gold nanostructures are directly linked to a gold thin film. Numerical simulations suggest that this LSP mode is dipolar and that the hot spots of the electric field are pushed to the top corners of the NPs, which makes it very sensitive to surrounding medium optical index changes and thus appealing for sensing applications. A figure of merit of such a system (gold/graphene/Au NPs) is 2.8, as compared to 2.1 for gold/Au NPs. This represents a 33 % gain in sensitivity and opens-up new sensing strategies.     

Biosynthesis of fluorescent gold nanoparticles using an edible freshwater red alga,Lemanea fluviatilis (L.) C.Ag. and antioxidant activity of biomatrix loaded nanoparticles

Biosynthesis of gold nanoparticles has been accomplished via reduction of an aqueous chloroauric acid solution with the dried biomass of an edible freshwater epilithic red alga, Lemanea fluviatilis (L.) C.Ag., as both reductant and stabilizer. The synthesized nanoparticles were characterized by UV–visible, powder X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier transform infrared (FT-IR), and dynamic light scattering (DLS) studies. The UV–visible spectrum of the synthesized gold nanoparticles showed the surface plasmon resonance (SPR) at around 530 nm. The powder XRD pattern furnished evidence for the formation of face-centered cubic structure of gold having average crystallite size 5.9 nm. The TEM images showed the nanoparticles to be polydispersed, nearly spherical in shape and have sizes in the range 5–15 nm. The photoluminescence spectrum of the gold nanoparticles excited at 300 nm showed blue emission at around 440 nm. Gold nanoparticles loaded within the biomatrix studied using a modified 2,2-diphenyl-1-picrylhydrazyl (DPPH) method exhibited pronounced antioxidant activity.     

Effects of Growth Solutions Ageing Time to the Formation of Gold Nanorods via Two-Step Approach for Plasmonic Applications

Plasmonics - We demonstrate the structural reorganization of gold nanorods (GNRs) that could fine-tune localized surface plasmon resonance (LSPR) by using modified wet chemical synthesis on the...     

The Effect of Aspect Ratio of Gold Nanorods on Cell Imaging with Two-Photon Excitation

To make the gold nanorod (AuNR) a better photoluminescence (PL) probe for cell imaging under two-photon excitation (TPE), the effect of the aspect ratio of AuNRs was studied. The AuNRs with the aspect ratios of 2.7, 3.2, 4.1, and 4.5 and correlated longitudinal surface plasmon resonance (LSPR) bands of 710, 760, 820, and 870 nm were compared. The approach of two-photon excited PL was used to measure the two-photon absorption cross section (TPACS) of these AuNRs in aqueous solutions. Under TPE of an 800-nm femtosecond laser, the TPACS of AuNRs with an aspect ratio of 3.2 was found to be the highest (about 3?×?10⁹ GM), and that of AuNRs (aspect ratio of 2.7) was only 1.5?×?10⁹ GM. The probe function of these two AuNRs was further compared in cell imaging studies using the human liver cancer cell (QGY) as the cell model. Both TPE PL image and confocal reflectance image of AuNR-loaded cells were acquired comparatively in measurements. The brightness and contrast of confocal reflectance images for these two AuNRs in cells are similar. In contrast, the PL images of cellular AuNRs (2.7) under TPE of 800 nm are weak but that of cellular AuNRs (3.2) is much better. These results show that when the LSPR band of AuNRs is coincided with the excitation wavelength, the TPACS of these AuNRs will be enhanced ensuring a good quality of cell imaging under TPE. The LSPR band is correlated to the aspect ratio of AuNRs. Therefore, in cell imaging studies with TPE, the aspect ratio effect of AuNRs should be taken into consideration.     

Plasmonic Biosensor Based on Triangular Au/Ag and Au/Ag/Au Core/Shell Nanoprisms onto Indium Tin Oxide Glass

Gold–silver core–shell triangular nanoprisms (Au/AgTNPs) were grown onto transparent indium tin oxide (ITO) thin film-coated glass substrate through a seed-mediated growth method without using peculiar binder molecules. The resulting Au/AgTNPs were characterized by scanning electron microscopy, atomic force microscopy, X-ray diffraction, UV–vis spectroscopy, and cyclic voltammograms. The peak of dipolar plasmonic resonance was located at near infrared region of ～700 nm, which showed the refractive index (RI) sensitivity of 248 nm/RIU. Moreover, thin gold shells were electrodeposited onto the surface of Au/AgTNPs in order to stabilize nanoparticles. Compared with the Au/AgTNPs, this peak of localized surface plasmon resonance (LSPR) was a little red-shift and decreased slightly in intensity. The refractive index sensitivity was estimated to be 287 nm/RIU, which showed high sensitivity as a LSPR sensing platform. Those triangular nanoprisms deposited on the ITO substrate could be further functionalized to fabricate LSPR biosensors. Results of this research show a possibility of improving LSPR sensor by using core–shell nanostructures.     

Obtain Quadruple Intense Plasmonic Resonances from Multilayered Gold Nanoshells by Silver Coating: Application in Multiplex Sensing

Four intense and separate localized surface plasmon resonance (LSPR) absorption peaks have been obtained in the gold-dielectric–gold–silver multilayer nanoshells. The silver coating on the gold shell results in a new LSPR peak at about 400 nm corresponding to the $ {{\left| {\omega_{+}^{-}} \right\rangle}_{Ag }} $ mode. The intense local electric field concentrated in the silver shell at the wavelength of 400 nm indicates that this new plasmonic band is coming from the symmetric coupling between the antibonding silver shell plasmon mode and the inner sphere plasmon. Increasing the silver shell thickness also leads to the intensity increasing of the $ {{\left| {\omega_{+}^{-}} \right\rangle}_{Au }} $ mode and blue shift of $ \left| {\omega_{-}^{+}} \right\rangle $ and $ \left| {\omega_{-}^{-}} \right\rangle $ modes. Therefore, quadruple intense plasmonic resonances in the visible region could be achieved in gold-dielectric–gold–silver multilayer nanoshells by tuning the geometrical parameters. And the quadruple intense plasmonic resonances in the visible region provide well potential for multiplex biosensing based on LSPR.     

Large Scale Fabrication of Gold Nano-Structured Substrates Via High Temperature Annealing and Their Direct Use for the LSPR Detection of Atrazine

The present work is reporting on the fabrication of localized surface plasmonic resonant (LSPR) gold nano-structures on glass substrate by using different high annealing temperatures (500 °C, 550 °C, 600 °C) of initially created semi-continue gold films (2 nm and 5 nm) by the electron beam evaporation technique. Interestingly, well-defined gold nano-structures were also obtained from continuous 8 nm evaporated gold film - known as the value above gold percolated thickness - once exposed to high temperatures. The surface morphology and plasmonic spectroscopy of “annealed” nano-structures were controlled by key experimental parameters such as evaporated film thickness and annealing temperature. By using scanning electron microscopy (SEM) characterization of annealed surface it was noticed that the size and inter-particle distance between nano-structures were highly dependent on the evaporated thin film thickness, while the nanoparticle shape evolution was mainly affected by the employed annealing temperature. Due to the well-controlled morphology of gold nano-particles, prominent and stable LSPR spectra were observed with good plasmon resonance tunability from 546 nm to 780 nm that recommend the developed protocol as a robust alternative to fabricate large scale LSPR surface. An example of a LSPR-immunosensor is reported. Thus, the monoclonal anti-atrazine antibodies immobilizion on the “annealed” gold nano-structures, as well as the specific antigen (atrazine) recognition were monitored as variations of the resonance wavelength shifts and optical density changes in the extinction measurements.     

Nanostructure shape effects on response of plasmonic aptamer sensors

A localized surface plasmon resonance (LSPR) sensor surface was fabricated by the deposition of gold nanorods on a glass substrate and subsequent immobilization of the DNA aptamer, which specifically bind to thrombin. This LSPR aptamer sensor showed a response of 6‐nm λ_max shift for protein binding with the detection limit of at least 10 pM, indicating one of the highest sensitivities achieved for thrombin detection by optical extinction LSPR. We also tested the LSPR sensor fabricated using gold bipyramid, which showed higher refractive index sensitivity than the gold nanorods, but the overall response of gold bipyramid sensor appears to be 25% less than that of the gold nanorod substrate, despite the approximately twofold higher refractive index sensitivity. XPS analysis showed that this is due to the low surface density of aptamers on the gold bipyramid compared with gold nanorods. The low surface density of the aptamers on the gold bipyramid surface may be due to the effect of shape of the nanostructure on the kinetics of aptamer monolayer formation. The small size of aptamers relative to other bioreceptors is the key to achieving high sensitivity by biosensors on the basis of LSPR, demonstrated here for protein binding. The generality of aptamer sensors for protein detection using gold nanorod and gold nanobipyramid substrates is anticipated to have a large impact in the important development of sensors toward biomarkers, environmental toxins, and warfare agents. Copyright © 2013 John Wiley & Sons, Ltd.     

Improvement of Figure of Merit for Gold Nanobar Array Plasmonic Sensors

We report a strategy to improve two types of the figure of merit (FOM and FOM*) of the refractive index sensitivity of a gold nanobar array localized surface plasmon resonance (LSPR) biosensor by simply placing it close to a thin gold film with a dielectric spacer. The thickness of the dielectric spacer determines the plasmon coupling strength between the gold nanobars and the gold film and consequently the FOM and FOM* of the biosensor. From our calculations, when the spacer thickness is 20 nm, the FOM and FOM* reach maximal (4.68 and 310, respectively) and the sensitivity remains at a high value of 600 nm per refractive index unit. This biosensor scheme is practically realizable, and this strategy is also potentially applicable to the LSPR biosensors with other geometries.     

Tailored Aggregate-Free Au Nanoparticle Decorations with Sharp Plasmonic Peaks on a U-Type Optical Fiber Sensor by Nanosecond Laser Irradiation

A novel experimental methodology is presented for fabricating U-shaped optical fiber probes decorated with aggregate-free Au nanoparticles exhibiting sharp localized surface plasmon resonance (LSPR) spectra. The U-type tip is coated with gold nanoparticles (AuNPs) using a simple and time-efficient dip-coating procedure, without initially taking any care to prevent the formation of nanoparticle aggregates in the coated area. In a second step, the coating was irradiated with a few tens of laser pulses of 5-ns duration at 532 nm with intensities in the range of 2–14 MW/cm², leading to the formation of aggregate-free LSPR optical fiber probes. The process was monitored and controlled in real time through the changes induced into the fiber’s extinction spectra by the laser irradiation, and the coated fibers were characterized by electron microscopy. The proposed methodology resulted into the fabrication of U-type optical fiber probes coated with AuNPs exhibiting a sharp plasmon peak, which is a perquisite for their application as sensing devices.     

Novel polyhedral gold nanoparticles: green synthesis,optimization and characterization by environmental isolate of Acinetobacter sp. SW30

Gold nanoparticles have enormous applications in cancer treatment, drug delivery and nanobiosensor due to their biocompatibility. Biological route of synthesis of metal nanoparticles are cost effective and eco-friendly. Acinetobacter sp. SW 30 isolated from activated sewage sludge produced cell bound as well as intracellular gold nanoparticles when challenged with HAuCl₄ salt solution. We first time report the optimization of various physiological parameters such as age of culture, cell density and physicochemical parameters viz HAuCl₄ concentration, temperature and pH which influence the synthesis of gold nanoparticles. Gold nanoparticles thus produced were characterized by various analytical techniques viz. UV–Visible spectroscopy, X-ray diffraction, cyclic voltammetry, transmission electron microscopy, selected area electron diffraction, high resolution transmission electron microscopy, environmental scanning electron microscopy, energy dispersive X-ray spectroscopy, atomic force microscopy and dynamic light scattering. Polyhedral gold nanoparticles of size 20 ± 10 nm were synthesized by 24 h grown culture of cell density 2.4 × 10⁹ cfu/ml at 50 °C and pH 9 in 0.5 mM HAuCl₄. It was found that most of the gold nanoparticles were released into solution from bacterial cell surface of Acinetobacter sp. at pH 9 and 50 °C.     

A Localized Surface Plasmon Resonance Biosensor Based on Integrated Controllable Au2S/AuAgS-Coated Gold Nanorods Composite

In this work, we have demonstrated that the exquisite optical properties based on localized surface plasmon resonance (LSPR) of Au₂S/AuAgS-coated gold nanorods (Au₂S/AuAgS-coated GNRs) can be utilized to develop a simple and sensitive biosensor, and goat anti-human IgG can be detected by the human IgG probe as low as 0.2 nM. Moreover, we introduce an integrated LSPR biosensor constructed by integrating Au₂S/AuAgS-coated GNRs immobilized on glass slide and isolated Au₂S/AuAgS-coated GNRs in the form of liquid. The detection of target binding was performed via direct spectral changes induced by changes of refractive index in the vicinity of individual particles. The integrated LSPR optical biosensor is label-free, cost-effective, and easy to fabricate and requires only a visible/near-infrared spectrometer for detection purposes. Additionally, the investigation on the mutual influence of the two types of nanorods in the integrated LSPR biosensor was performed. The results of separate experiments indicate that the nanorods in the form of isolate or in integrated exhibit a similar behavior.     

Speckled SiO2@Au Core–Shell Particles as Surface Enhanced Raman Scattering Probes

Silica particles of ~800 nm size were functionalized using 3-amino propyl triethoxysilane molecules on which gold particles (~20 nm size) were deposited. The resulting particles appeared to form speckled SiO₂@Au core–shell particles. The surface roughness, along with hot spots, due to nanogaps between the gold nanoparticles was responsible for the enhancement of the Raman signal of crystal violet molecules by ~3.2?×?10⁷ and by ~1.42?×?10⁸ of single-wall carbon nanotubes. It has also been observed that the electromagnetic excitation near surface plasmon resonance (SPR) of core–shell particles is more effective than off resonance SPR excitation.     

Synthesis of Silver and Gold Nanoparticles Using Cashew Nut Shell Liquid and Its Antibacterial Activity Against Fish Pathogens

This study reveals a green process for the production of multi-morphological silver (Ag NPs) and gold (Au NPs) nanoparticles, synthesized using an agro-industrial residue cashew nut shell liquid. Aqueous solutions of Ag⁺ ions for silver and chloroaurate ions for gold were treated with cashew nut shell extract for the formation of Ag and Au NPs. The nano metallic dispersions were characterized by measuring the surface plasmon absorbance at 440 and 546 nm for Ag and Au NPs. Transmission electron microscopy showed the formation of nanoparticles in the range of 5–20 nm for silver and gold with assorted morphologies such as round, triangular, spherical and irregular. Scanning electron microscopy with energy dispersive spectroscopy and X-ray diffraction analyses of the freeze-dried powder confirmed the formation of metallic Ag and Au NPs in crystalline form. Further analysis by Fourier transform infrared spectroscopy provided evidence for the presence of various biomolecules, which might be responsible for the reduction of silver and gold ions. The obtained Ag and Au NPs had significant antibacterial activity, minimum inhibitory concentration and minimum bactericidal concentration on bacteria associated with fish diseases.     

Gold Nanoring Arrays for Near Infrared Plasmonic Biosensing

Gold nanoring array surfaces that exhibit strong localized surface plasmon resonances (LSPR) at near infrared (NIR) wavelengths from 1.1 to 1.6 μm were used as highly sensitive real-time refractive index biosensors. Arrays of gold nanorings with tunable diameter, width, and spacing were created by the nanoscale electrodeposition of gold nanorings onto lithographically patterned nanohole array conductive surfaces over large areas (square centimeters). The bulk refractive index sensitivity of the gold nanoring arrays was determined to be up to 3,780 cm⁻¹/refractive index unit by monitoring shifts in the LSPR peak by FT-NIR transmittance spectroscopy measurements. As a first application, the surface polymerization reaction of dopamine to form polydopamine thin films on the nanoring sensor surface from aqueous solution was monitored with the real-time LSPR peak shift measurements. To demonstrate the utility of the gold nanoring arrays for LSPR biosensing, the hybridization adsorption of DNA-functionalized gold nanoparticles onto complementary DNA-functionalized gold nanoring arrays was monitored. The adsorption of DNA-modified gold nanoparticles onto nanoring arrays modified with mixed DNA monolayers that contained only 0.5 % complementary DNA was also detected; this relative surface coverage corresponds to the detection of DNA by hybridization adsorption from a 50 pM solution.

     

Spectral Characteristics of Near-Infrared Surface Plasmon Resonance

Intrinsic properties of surface plasmons (SPs) excited with Kretschmann configuration were analyzed as a function of wavelength, including the propagation length, the penetration depth, the Goos–Hänchen (GH) shift, and the field enhancement. The calculated results indicate that there exists a critical thickness (t _cr) of the gold layer and that the maximum GH shift occurring exactly at the SP resonance wavelength (λ _R) rapidly varies from positive to negative with changing of the gold layer thickness from t?<?t _cr to t?>?t _cr. The maximum field enhancement happens not at λ _R but at a wavelength smaller than λ _R due to the phase retardation between the transmitted and reflected light. Simulations also reveal that a broadband collimated near-infrared beam can simultaneously excite two SPs with different responses to a refractive index (RI) change: the shorter-wavelength SP able to make a small redshift and the longer-wavelength SP capable of yielding a large blueshift. Only the shorter-wavelength SP was experimentally observed and its RI sensitivity was measured to increase from 3,539 nm/RIU at λ _R?=?707.6 nm to 57,143 nm/RIU at λ _R?=?1,398 nm. The SP at λ _R?=?1,013 nm moved to λ _R?=?1,029 nm in response to the saturation adsorption of bovine serum albumin, and the corresponding surface coverage was determined to be Γ?=?1.565 ng/mm² based on a quasilinear dependence of Γ on the resonance wavelength shift (?λ _R) deduced theoretically. Butyrylcholinesterase adsorption from a dilute solution of 10 nM protein in phosphate buffer solution leads to a redshift of ?λ _R?=?10 nm, corresponding to Γ?≈?0.97 ng/mm².     

Search of Extremely Sensitive Near-Infrared Plasmonic Interfaces: A Theoretical Study

The sensitivity of the wavelength position of localized surface plasmon resonance (LSPR) in metal nanostructures to local changes in the refractive index has been widely used for label-free detection strategies. Tuning the optical properties of the nanostructures from the visible to the infrared region is expected to have a drastic effect on the refractive index sensitivity. Here, we theoretically investigate the optical response of a newly designed plasmonic interface to changes in the bulk refractive index by the finite difference time domain method. It consists of a structured interface, where the planar interface is superposed with dielectric pillars 30 nm in height and 125 nm in length with a separation distance of 15 nm. The pillars are covered with U-shaped gold nanostructures of 50 nm in height, 125 nm in length, and 5 nm of gold base thickness. The whole structure is finally covered with a 5-nm thick dielectric layer of n ₂?=?2.63. This plasmonic structure shows bulk refractive index sensitivities up to 1750 nm/RIU (RIU : refractive index unit) in the near infrared (λ?=?2621 nm). The enhanced sensitivity is a consequence of the extremely enhanced electrical field between the gold nanopillars of the plasmonic interface.     

Localized surface plasmon resonance based gold nanobiosensor: Determination of thyroid stimulating hormone

An immunoassay method based on the peak shift of the localized surface plasmon resonance (LSPR) absorption maxima has been developed for the determination of the thyroid stimulating hormone (TSH) in human blood serum. The anti-TSH antibody was adsorbed on the synthesized gold nanoparticles by electrostatic forces. The efficiency of the nanobiosensor was improved by optimizing the factors affecting the probe construction such as the pH and the antibody to gold nanoparticles ratio. Dynamic light scattering was applied for the characterization of the constructed probe. The amount of peak shift of the LSPR absorption maxima was selected as the basis for determination of TSH antigen. The linear dynamic range of 0.4–12.5 mIU L⁻¹ and the calibration sensitivity of 1.71 L mIU⁻¹ were obtained. The human control serum sample was analyzed for TSH by constructed nanobiosensor and the acceptable results were obtained.     

Interaction of dimeric horse cytochrome c with cyanide ion

We have previously shown that methionine–heme iron coordination is perturbed in domain-swapped dimeric horse cytochrome c. To gain insight into the effect of methionine dissociation in dimeric cytochrome c, we investigated its interaction with cyanide ion. We found that the Soret and Q bands of oxidized dimeric cytochrome c at 406.5 and 529 nm redshift to 413 and 536 nm, respectively, on addition of 1 mM cyanide ion. The binding constant of dimeric cytochrome c and cyanide ion was obtained as 2.5 × 10⁴ M^?1. The Fe–CN and C–N stretching (ν _Fe–CN and ν _CN) resonance Raman bands of CN^?-bound dimeric cytochrome c were observed at 443 and 2,126 cm^?1, respectively. The ν _Fe–CN frequency of dimeric cytochrome c was relatively low compared with that of other CN^?-bound heme proteins, and a relatively strong coupling between the Fe–C–N bending and porphyrin vibrations was observed in the 350–450-cm^?1 region. The low ν _Fe–CN frequency suggests weaker binding of the cyanide ion to dimeric cytochrome c compared with other heme proteins possessing a distal heme cavity. Although the secondary structure of dimeric cytochrome c did not change on addition of cyanide ion according to circular dichroism measurements, the dimer dissociation rate at 45 °C increased from (8.9 ± 0.7) × 10^?6 to (3.8 ± 0.2) × 10^?5 s^?1, with a decrease of about 2 °C in its dissociation temperature obtained with differential scanning calorimetry. The results show that diatomic ligands may bind to the heme iron of dimeric cytochrome c and affect its stability.