Fluorophore-Decorated Mie Resonant Silicon Nanosphere for Scattering/Fluorescence Dual-Mode Imaging

Inorganic nanoparticles with multiple functions have been attracting attention as multimodal nanoprobes in bioimaging, biomolecule detection, and medical diagnosis and treatment. A drawback of conventional metallic nanoparticle-based nanoprobes is the Ohmic losses that lead to fluorescence quenching of attached molecules and local heating under light irradiation. Here, metal-free nanoprobes capable of scattering/fluorescence dual-mode imaging are developed. The nanoprobes are composed of a silicon nanosphere core having efficient Mie scattering in the visible to near infrared range and a fluorophore doped silica shell. The dark-field scattering and photoluminescence images/spectra for nanoprobes made from different size silicon nanospheres and different kinds of fluorophores are studied by single particle spectroscopy. The fluorescence spectra are strongly modified by the Mie modes of a silicon nanosphere core. By comparing scattering and fluorescence spectra and calculated Purcell factors, the fluorescence enhancement factor is quantitatively discussed. In vitro scattering/fluorescence imaging studies on human cancer cells demonstrate that the developed nanoparticles work as scattering/fluorescence dual-mode imaging nanoprobes.     

Biological imaging of HEK293 cells expressing PLCgamma1 using surface-enhanced Raman microscopy

Surface-enhanced Raman scattering (SERS) imaging has been used for the targeting and imaging of specific cancer markers in live cells. For this purpose, Au/Ag core-shell nanoparticles, conjugated with monoclonal antibodies, were prepared. The procedures to label live cells with those bimetallic nanoprobes have been developed and used for highly sensitive SERS imaging of live cells. In the present study, live HEK293 cells expressing PLCgamma1 have been used as the optical imaging target. Our results demonstrate the potential feasibility of SERS imaging technology for the highly sensitive imaging of cancer biomarkers in live cells.     

Gold and magnetic oxide/gold core/shell nanoparticles as bio-functional nanoprobes

The ability to create bio-functional nanoprobes for the detection of biological reactivity is important for developing bioassay and diagnostic methods. This paper describes the findings of an investigation of the surface functionalization of gold (Au) and magnetic nanoparticles coated with gold shells (M/Au) by proteins and spectroscopic labels for the creation of nanoprobes for use in surface enhanced Raman scattering (SERS) assays. Highly monodispersed Au nanoparticles and M/Au nanoparticles with two types of magnetic nanoparticle cores (Fe(2)O(3) and MnZn ferrite) were studied as model systems for the bio-functionalization and Raman labeling. Comparison of the SERS intensities obtained with different particle sizes (30-100?nm) and samples in solution versus on solid substrates have revealed important information about the manipulation of the SERS signals. In contrast to the salt-induced uncontrollable and irreversible aggregation of nanoparticles, the ability to use a centrifugation method to control the formation of stable small clustering sizes of nanoparticles was shown to enhance SERS intensities for samples in solution as compared with samples on solid substrates. A simple method for labeling protein-capped Au nanoparticles with Raman-active molecules was also described. The functionalized Au and M/Au nanoparticles are shown to exhibit the desired functional properties for the detection of SERS signals in the magnetically separated reaction products. These results are discussed in terms of the interparticle distance dependence of 'hot-spot' SERS sites and the delineation of the parameters for controlling the core-shell reactivity of the magnetic functional nanocomposite materials in bio-separation and spectroscopic probing.     

Reaction Kinetics‐Mediated Control over Silver Nanogap Shells as Surface‐Enhanced Raman Scattering Nanoprobes for Detection of Alzheimer's Disease Biomarkers

It is very challenging to accurately quantify the amounts of amyloid peptides Aβ40 and Aβ42, which are Alzheimer's disease (AD) biomarkers, in blood owing to their low levels. This has driven the development of sensitive and noninvasive sensing methods for the early diagnosis of AD. Here, an approach for the synthesis of Ag nanogap shells (AgNGSs) is reported as surface‐enhanced Raman scattering (SERS) colloidal nanoprobes for the sensitive, selective, and multiplexed detection of Aβ40 and Aβ42 in blood. Raman label chemicals used for SERS signal generation modulate the reaction rate for AgNGSs production through the formation of an Ag‐thiolate lamella structure, enabling the control of nanogaps at one nanometer resolution. The AgNGSs embedded with the Raman label chemicals emit their unique SERS signals with a huge intensity enhancement of up to 10⁷ and long‐term stability. The AgNGS nanoprobes, conjugated with an antibody specific to Aβ40 or Aβ42, are able to detect these AD biomarkers in a multiplexed manner in human serum based on the AgNGS SERS signals. Detection is possible for amounts as low as 0.25 pg mL^?1. The AgNGS nanoprobe‐based sandwich assay has a detection dynamic range two orders of magnitude wider than that of a conventional enzyme‐linked immunosorbent assay.     

Synthesis and characterization of a disulfide reporter molecule for enhancing pH measurements based on surface-enhanced Raman scattering

In this paper, we describe the synthesis and characterization of 2,5-dimercaptobenzoic acid as a novel pH-sensitive disulfide reporter molecule for surface-enhanced Raman scattering (SERS) capable of inducing the controlled aggregation of gold (Au) colloids in solution without the addition of salts. While weak acids have been shown to yield some pH sensitivity as reporter molecules for SERS measurements, the reproducibility and signal strength of nanoparticle probes based on such molecules can vary greatly. This limited reproducibility depends greatly on the salt-induced aggregation of the colloidal nanoprobes, which is required in order to obtain SERS signals strong enough to probe individual clusters. This complicates their use in live cell sensing applications. We show that our approach results in primarily bridged nanoparticles comprising a pH-sensitive nanoprobe that can quantify accurately pH values well below 5.5. The robustness and sensitivity of this system makes it a powerful tool for measuring pH values on the nanoscale under in vitro conditions.     

Improving nanoprobes using surface-enhanced Raman scattering from 30-nm hollow gold particles

We report the development of nanoprobes that exploit the surface-enhanced Raman scattering (SERS) from nonaggregated, hollow, gold nanospheres (HGNSs). The homogeneity of the HGNSs leads to a nearly 10-fold improvement in signal consistency over standard silver SERS substrates, which translates into a significant increase in sensitivity and dynamic range for the model application of pH sensing. Moreover, the small size (30-nm diameter) of these SERS-active nanoparticles represents a major step in advancing sensing technology based on SERS, making this technology more amenable to intracellular sensing.     

High Surface‐Enhanced Raman Scattering Performance of Individual Gold Nanoflowers and Their Application in Live Cell Imaging

Molecular imaging techniques based on surface‐enhanced Raman scattering (SERS) face a lack of reproducibility and reliability, thus hampering its practical application. Flower‐like gold nanoparticles have strong SERS enhancement performance due to having plenty of hot‐spots on their surfaces, and this enhancement is not dependent on the aggregation of the particles. These features make this kind of particle an ideal SERS substrate to improve the reproducibility in SERS imaging. Here, the SERS properties of individual flower‐like gold nanoparticles are systematically investigated. The measurements reveal that the enhancement of a single gold nanoparticle is independent of the polarization of the excitation laser with an enhancement factor as high as 10⁸. After capping with Raman signal molecules and folic acid, the gold nanoflowers show strong Raman signal in the living cells, excellent targeting properties, and a high signal‐to‐noise ratio for SERS imaging.     

Arginine-glycine-aspartic acid-conjugated dendrimer-modified quantum dots for targeting and imaging melanoma

Angiogenesis is essential for the development of malignant tumors and provides important targets for tumor diagnosis and therapy. Quantum dots have been broadly investigated for their potential application in cancer molecular imaging. In present work, CdSe quantum dots were synthesized, polyamidoamine dendrimers were used to modify surface of quantum dots and improve their solubility in water solution. Then, dendrimer-modified CdSe quantum dots were conjugated with arginine-glycine-aspartic acid (RGD) peptides. These prepared nanoprobes were injected into nude mice loaded with melanoma (A375) tumor xenografts via tail vessels, IVIS imaging system was used to image the targeting and bio-distribution of as-prepared nanoprobes. The dendrimer-modified quantum dots exhibit water-soluble, high quantum yield, and good biocompatibility. RGD-conjugated quantum dots can specifically target human umbilical vein endothelial cells (HUVEC) and A375 melanoma cells, as well as nude mice loaded with A735 melanoma cells. High-performance RGD-conjugated dendrimers modified quantum dot-based nanoprobes have great potential in application such as tumor diagnosis and therapy.     

Split‐GFP: SERS Enhancers in Plasmonic Nanocluster Probes

The assembly of plasmonic metal nanoparticles into hot spot surface‐enhanced Raman scattering (SERS) nanocluster probes is a powerful, yet challenging approach for ultrasensitive biosensing. Scaffolding strategies based on self‐complementary peptides and proteins are of increasing interest for these assemblies, but the electronic and the photonic properties of such hybrid nanoclusters remain difficult to predict and optimize. Here, split‐green fluorescence protein (sGFP) fragments are used as molecular glue and the GFP chromophore is used as a Raman reporter to assemble a variety of gold nanoparticle (AuNP) clusters and explore their plasmonic properties by numerical modeling. It is shown that GFP seeding of plasmonic nanogaps in AuNP/GFP hybrid nanoclusters increases near‐field dipolar couplings between AuNPs and provides SERS enhancement factors above 10⁸. Among the different nanoclusters studied, AuNP/GFP chains allow near‐infrared SERS detection of the GFP chromophore imidazolinone/exocyclic C?C vibrational mode with theoretical enhancement factors of 10⁸–10⁹. For larger AuNP/GFP assemblies, the presence of non‐GFP seeded nanogaps between tightly packed nanoparticles reduces near‐field enhancements at Raman active hot spots, indicating that excessive clustering can decrease SERS amplifications. This study provides rationales to optimize the controlled assembly of hot spot SERS nanoprobes for remote biosensing using Raman reporters that act as molecular glue between plasmonic nanoparticles.     

Detection of human immunodeficiency virus type 1 DNA sequence using plasmonics nanoprobes

This paper describes the use of plasmonics-based nanoprobes that act as molecular sentinels for DNA diagnostics. The plasmonics nanoprobe comprises a metal nanoparticle and a stem-loop DNA molecule tagged with a Raman label. The nanoprobe utilizes the specificity and selectivity of the DNA hairpin probe sequence to detect a specific target DNA sequence of interest. In the absence of target DNA, the stem-loop configuration maintains the Raman label in proximity to the metal nanoparticle, inducing an intense surface-enhanced Raman scattering (SERS) effect that produces a strong Raman signal upon laser excitation. Upon hybridization of a complementary target DNA sequence to the nanoprobe, the stem-loop configuration is disrupted, causing the Raman label to physically separate from the metal nanoparticle, thus quenching the SERS signal. The usefulness and potential application of the plasmonics nanoprobe for diagnosis is demonstrated using the gag gene sequence of the human immunodeficiency virus type 1 (HIV-1). We successfully demonstrated the specificity and selectivity of the plasmonics nanoprobes to detect PCR amplicons of the HIV gene. The potential for combining the spectral selectivity and high sensitivity of the SERS process with inherent molecular specificity of DNA hairpins to diagnose molecular target sequences in homogeneous solutions is discussed.     

In vivo MRI detection of gliomas by chlorotoxin-conjugated superparamagnetic nanoprobes

Converging advances in the development of nanoparticle-based imaging probes and improved understanding of the molecular biology of brain tumors offer the potential to provide physicians with new tools for the diagnosis and treatment of these deadly diseases. However, the effectiveness of promising nanoparticle technologies is currently limited by insufficient accumulation of these contrast agents within tumors. Here a biocompatible nanoprobe composed of a poly(ethylene glycol) (PEG) coated iron oxide nanoparticle that is capable of specifically targeting glioma tumors via the surface-bound targeting peptide, chlorotoxin (CTX), is presented. The preferential accumulation of the nanoprobe within gliomas and subsequent magnetic resonance imaging (MRI) contrast enhancement are demonstrated in vitro in 9L cells and in vivo in tumors of a xenograft mouse model. TEM imaging reveals that the nanoprobes are internalized into the cytoplasm of 9L cells and histological analysis of selected tissues indicates that there are no acute toxic effects of these nanoprobes. High targeting specificity and benign biological response establish this nanoprobe as a potential platform to aid in the diagnosis and treatment of gliomas and other tumors of neuroectodermal origin.     

Multilayer enhanced gold film over nanostructure surface-enhanced Raman substrates

We have developed a novel class of gold multilayer, surface-enhanced Raman scattering (SERS) substrates that are capable of enhancing SERS signals by 15.3-fold over conventional gold film over nanostructure (GFON) SERS substrates, making them comparable in sensitivity to optimized silver film over nanostructure (SFON) substrates, while providing the long-term stability obtained from gold. They are fabricated by depositing 10 A thick silver oxide islands on conventional GFON substrates, followed by deposition of a second continuous gold layer. The silver oxide layer acts as a dielectric spacer between the two continuous gold films and produces significantly enhanced SERS signals, as compared to optimized single layer substrates of the same geometry or comparable substrates prepared by deposition of silver islands that are not oxidized. In addition to the enhanced sensitivity of these multilayer substrates, they also exhibit long SERS active shelf-lives (i.e., months), with no measurable degradation in SERS enhancement, and relative standard deviations in SERS enhancement of less than 5.2% across the substrate's surface.     

Competitive Reaction Pathway for Site‐Selective Conjugation of Raman Dyes to Hotspots on Gold Nanorods for Greatly Enhanced SERS Performance

Common methods to prepare SERS (surface‐enhanced Raman scattering) probes rely on random conjugation of Raman dyes onto metal nanostructures, but most of the Raman dyes are not located at Raman‐intense electromagnetic hotspots thus not contributing to SERS enhancement substantially. Herein, a competitive reaction between transverse gold overgrowth and dye conjugation is described to achieve site selective conjugation of Raman dyes to the hotspots (ends) on gold nanorods (GNRs). The preferential overgrowth on the nanorod side surface creates a barrier to prevent the Raman dyes from binding to the side surface except the ends of the GNRs, where the highest SERS enhancement factors are expected. The SERS enhancement observed from this special structure is dozens of times larger than that from conjugates synthesized by conventional methods. This simple and powerful strategy to prepare SERS probes can be extended to different anisotropic metal nanostructures with electromagnetic hotspots and has immense potential in in‐depth SERS‐based biological imaging and single‐molecule detection.     

New surface-enhanced Raman spectroscopy substrates via self-assembly of silver nanoparticles for perchlorate detection in water

Perchlorate (ClO4-) has recently emerged as a widespread contaminant in drinking water and groundwater supplies in the United States, and a need exists for rapid detection and monitoring of this contaminant. In this study, surface-enhanced Raman spectroscopy (SERS) was studied as a means of ClO4- detection, and new sol-gel-based SERS substrates were developed by self-assembly of silver colloidal nanoparticles with various functionalized silane reagents. These substrate materials were tailored to allow detection of ClO4- in water with improved sorptivity, stability, and sensitivity and with the ability to detect ClO4- at concentrations as low as 10(-6) M (or 100 microg/L) with good reproducibility. Similar techniques were used to fabricate capillary SERS flow cells by assembling functionalized silver nanoparticles capable of attracting ClO4- to the SERS surface or the internal wall of glass capillaries. These capillary flow cells could be readily configured to allow for in situ, nondestructive detection of ClO4- via fiber optics.     

Synthesis and characterisation of iron oxide nanoparticles conjugated with epidermal growth factor receptor (EGFR) monoclonal antibody as MRI contrast agent for cancer detection

The aim of this study is to synthesise superparamagnetic iron oxide nanoparticles conjugated with anti‐epidermal growth factor receptor monoclonal antibody (ANTI‐EGFR‐SPION) and investigate its physicochemical characterisation and biocompatibility as a targeted magnetic resonance imaging (MRI) contrast agent for the EGFR‐specific detection in EGFR expressing tumour cells. These particles employed biocompatible polymers, poly(D,L‐lactide‐co‐glycolide) (PLGA) and polyethylene glycol aldehyde (PEG‐aldehyde), to increase the half‐life of particles in circulation and reduce their side effects. The Fe₃ O₄ ‐loaded PLGA‐PEG‐aldehyde nanoparticles were prepared by a modified water‐in‐oil‐in‐water double emulsion method. The EGFR antibody was conjugated to the surface of SPIONs using the aldehyde‐amine reaction. Synthesised conjugates (nanoprobes) were characterised using Fourier transform infrared spectrophotometry, dynamic light scattering, transmission electron microscopy images, and vibrating‐sample magnetometery, and the results showed that the conjugation was successful. The mean diameter of nanoprobes was about 25 nm. These nanoprobes exhibited excellent water‐solubility, stability, and biocompatibility. Meanwhile, MR susceptibility test proved that synthesised nanoprobes can be managed for negative contrast enhancement. The results of this study suggested the potential use of these nanoprobes for non‐invasive molecular MRI in EGFR detection in the future.Inspec keywords: solubility, nanomedicine, cancer, spectrophotometry, emulsions, biomedical MRI, nanomagnetics, nanofabrication, tumours, nanoparticles, magnetic particles, molecular biophysics, light scattering, proteins, cellular biophysics, Fourier transform spectra, superparamagnetism, polymers, transmission electron microscopy, iron compoundsOther keywords: physicochemical characterisation, superparamagnetic iron oxide nanoparticles, novel targeting cancer detection, anti‐epidermal growth factor receptor monoclonal antibody, ANTI‐EGFR‐SPION, biocompatibility, targeted magnetic resonance imaging contrast agent, EGFR‐specific detection, EGFR expressing tumour cells, biocompatible polymers, PLGA‐PEG‐aldehyde nanoparticles, modified water‐in‐oil‐in‐water double emulsion method, EGFR antibody, aldehyde‐amine reaction, synthesised conjugates were characterised using Fourier, transmission electron microscopy images, synthesised nanoprobes, EGFR detection, size 25.0 nm, Fe₃ O₄      

Smart nanoprobes for ultrasensitive detection of breast cancer via magnetic resonance imaging

Antibody-conjugated hydrophilic magnetic nanocrystals for use as smart nanoprobes were developed for ultrasensitive detection of breast cancer via magnetic resonance (MR) imaging. MnFe(2)O(4) nanocrystals (MNCs) for use as MR imaging contrast agents were synthesized by thermal decomposition to take advantage of their MR signal enhancement effect. The MNC surfaces were then modified with amphiphilic tri-block copolymers (dicarboxy poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol)), not only allowing the MNCs to transfer from the organic to the aqueous phase, but also increasing the colloidal stability of the MNCs by masking poly(ethylene glycol). The physicochemical properties of the synthesized hydrophilic magnetic nanocrystals (HMNCs) were fully investigated. Trastuzumab (TZ), a monoclonal antibody against human epidermal growth factor receptor (HER2/neu), was further conjugated on the surface of HMNCs to specifically target HER2/neu over-expressed breast cancer cells. MR imaging analysis of target cells treated with TZ-conjugated HMNCs (TZ-HMNCs) clearly demonstrated their potential as high-performance nanoprobes for selective imaging.     

Development of optical nanoprobes for molecular imaging of reactive oxygen and nitrogen species

Reactive oxygen and nitrogen species (RONS) play important roles in cell signal transduction. However, overproduction of RONS is associated with a series of pathological processes and may disrupt cellular homeostasis, causing oxidative and nitrosative stress. Accurate methods to selectively and specifically monitor RONS in living systems are required to further elucidate the biological functions of these species. Optical imaging possesses high sensitivity, high spatiotemporal resolution, and real-time imaging capability. These qualities are advantageous for the detection of RONS in living systems. This review summarizes the development of optical nanoprobes with near-infrared (NIR) fluorescent, upconversion luminescent, chemiluminescent, or photoacoustic signals for molecular imaging of RONS in living systems. In this review, we discuss the design principles and advantages of RONS-responsive activatable nanoprobes, as well as applications of these optical imaging modalities in different disease models.     

Ag nanorod arrays tailored for surface-enhanced Raman imaging in the near-infrared region

Using a dynamic oblique angle deposition technique, we demonstrate the direct formation of Ag nanorods with quasi-parallel major axes on a template layer of oxide having a strongly anisotropic surface morphology. The optical properties of the nanorods are tuned by varying the deposition conditions without any pre-?or post-treatment, and the resulting Ag nanorod arrays exhibit high surface-enhanced Raman scattering (SERS) activity. In addition to high macroscopic uniformity over a large area, our nanorod arrays contain a high density of isolated nanorods. Using the optimum Ag nanorod arrays, the SERS imaging of the microdroplets of a rhodamine 6G solution is successfully demonstrated. The space resolution of the imaging is of the order of at least a few μm. These features are suitable for the SERS imaging of biomaterials.     

Characterization of silane-modified immobilized gold colloids as a substrate for surface-enhanced Raman spectroscopy

Immobilized gold colloid particles coated with a C-18 alkylsilane layer have been characterized as a substrate for surface-enhanced Raman scattering (SERS) studies of adsorption onto hydrophobic surfaces. Atomic force microscopy images, optical extinction spectra, and SERS measurements are reported as a function of accumulation of gold colloid on glass. As the metal particles become increasingly aggregated on the surface, the SERS enhancement increases until the plasmon resonance shifts to wavelengths longer than the excitation laser. The gold colloid substrates are stable and exhibit reproducible SERS enhancement. When octadecyltrimethoxysilane is self-assembled over the gold, the metal surface is protected from exposure to solution-phase species, as evidenced by the inhibition of chemisorption of a disulfide reagent to the overcoated gold surface. The results show that interactions with gold can be blocked by a silane layer so as not to significantly influence physisorption of molecules at the C-18/solution interface. The SERS enhancement from these C-18-overcoated gold substrates is reproducible for different films prepared from the same colloidal suspension; the substrates are also stable with time and upon exposure to laser irradiation.     

Plasmonic coupling interference (PCI) nanoprobes for nucleic acid detection

A label-free approach using plasmonic coupling interference (PCI) nanoprobes for nucleic acid detection using surface-enhanced Raman scattering (SERS) is described. To induce a strong plasmonic coupling effect, a nanonetwork of silver nanoparticles with the Raman label located between adjacent nanoparticles is assembled by Raman-labeled DNA-locked nucleic acid (LNA) duplexes. The PCI method then utilizes specific nucleic acid sequences of interest as competitor elements for the Raman-labeled DNA strands to interfere the formation of nanonetworks in a competitive binding process. As a result, the plasmonic coupling effect induced through the formation of the nanonetworks is significantly diminished, resulting in a reduced SERS signal. The potential of the PCI technique for biomedical applications is illustrated by detecting single-nucleotide polymorphism (SNP) and microRNA sequences involved in breast cancers. The results of this study could lead to the development of nucleic acid diagnostic tools for biomedical diagnostics and biosensing applications using SERS detection.