Resonance energy transfer-amplifying fluorescence quenching at the surface of silica nanoparticles toward ultrasensitive detection of TNT

This paper reports a resonance energy transfer-amplifying fluorescence quenching at the surface of silica nanoparticles for the ultrasensitive detection of 2,4,6-trinitrotoluene (TNT) in solution and vapor environments. Fluorescence dye and organic amine were covalently modified onto the surface of silica nanoparticles to form a hybrid monolayer of dye fluorophores and amine ligands. The fluorescent silica particles can specifically bind TNT species by the charge-transfer complexing interaction between electron-rich amine ligands and electron-deficient aromatic rings. The resultant TNT-amine complexes bound at the silica surface can strongly suppress the fluorescence emission of the chosen dye by the fluorescence resonance energy transfer (FRET) from dye donor to the irradiative TNT-amine acceptor through intermolecular polar-polar interactions at spatial proximity. The quenching efficiency of the hybrid nanoparticles with TNT is greatly amplified by at least 10-fold that of the corresponding pure dye. The nanoparticle-assembled arrays on silicon wafer can sensitively detect down to approximately 1 nM TNT with the use of only 10 microL of solution (approximately 2 pg TNT) and several ppb of TNT vapor in air. The simple FRET-based nanoparticle sensors reported here exhibit a high and stable fluorescence brightness, strong analyte affinity, and good assembly flexibility and can thus find many applications in the detection of ultratrace analytes.     

Alkyl-functionalized oxide-free silicon nanoparticles: synthesis and optical properties

Highly monodisperse silicon nanoparticles (1.57 +/- 0.21 nm) are synthesized with a covalently attached alkyl monolayer on a gram scale. Infrared spectroscopy shows that these silicon nanoparticles contain only a few oxygen atoms per nanoparticle. XPS spectra clearly show the presence of unoxidized Si and attached alkyl chains. Owing to the relatively efficient synthesis (yields approximately 100-fold higher than of those previously reported) the molar extinction coefficient epsilon can be measured: epsilon(max) = 1.7 x 10(-4) M(-1)cm(-1), only a factor of 4 lower than that of CdS and CdSe nanoparticles of that size. The quantum yield of emission ranges from 0.12 (C(10)H(21)-capping) to 0.23 (C(16)H(33)-capping). UV/Vis absorption and emission spectroscopy show clear vibrational progressions (974 +/- 14 cm(-1); up to five vibrational bands visible at room temperature), resembling bulk SiC phonons, which support the monodispersity observed by TEM. This was also confirmed by time-resolved fluorescence anisotropy measurements, which display a strictly monoexponential decay that can only be indicative of monodisperse, ball-shaped nanoparticles.     

Single molecule spectroscopy of organic dye nanoparticles

Organic dye nanoparticles 1-13 nm in height and 10-45 nm in width were prepared by the reprecipitation method. With single-molecule/nanoparticle spectroscopy, two distinct types of nanoparticles were found: particles with blue emission and particles with red emission. The difference in spectral characteristics is attributed to the presence of two morphological types of particles in the samples. The presence of two types of nanoparticles in the samples was further corroborated by our ability to separate blue nanoparticles from red nanoparticles by centrifugation.     

Dependence of fluorescence intensity on the spectral overlap between fluorophores and plasmon resonant single silver nanoparticles

We investigate the fluorescence from dyes coupled to individual DNA-functionalized metal nanoparticles. We use single-particle darkfield scattering and fluorescence microscopy to correlate the fluorescence intensity of the dyes with the localized surface plasmon resonance (LSPR) spectra of the individual metal nanoparticles to which they are attached. For each of three different dyes, we observe a strong correlation between the fluorescence intensity of the dye and the degree of spectral overlap with the plasmon resonance of the nanoparticle. On average, we observe the brightest fluorescence from dyes attached to metal nanoparticles that have a LSPR scattering peak approximately 40-120 meV higher in energy than the emission peak of the fluorophore. These results should prove useful for understanding and optimizing metal-enhanced fluorescence.     

Tuning luminescence intensity of RHO6G dye using silver nanoparticles

The photoluminescence (PL) from rhodamine (RHO6G) dye dispersed in ethanol has been studied in the presence of different amounts of citrate stabilized silver nanoparticles of size, ∼10 nm. Enhancement as well as quenching of luminescence intensity has been observed and it was found that luminescence intensity can be tuned by adding various amounts of silver nanoparticles to the RHO6G dye dispersion. The luminescence spectra of dye consist of two peaks at 440 nm and 550 nm. Peak at 440 nm shows an enhancement in intensity at all the concentrations of added silver nanoparticles with the maximum intensity for dye with 0.25 ml silver nanoparticles (82% enhancement in the luminescence intensity). PL intensity of intense peak at 550 nm of dye molecules was found to be quenched in presence of silver nanoparticles and maximum quenching was found to be 41% for the dye with 1 ml silver nanoparticles. However, for lowest concentration of silver nanoparticles viz. (0.01 ml), enhancement in intensity was observed (13% enhancement than the dye molecules). The quenching as well as enhancement in the intensity can be understood by considering the possibility of three different phenomena. It has been reported earlier that when metal nanoparticles are in close proximity to the fluorophores, quenching of luminescence occurs, whereas when metal nanoparticles are located at certain distance, enhancement in luminescence is observed. This effect has been explained by coupling of surface plasmon resonance from metal nanoparticles with fluorophore, resulting in the increase of excitation and emission rate of the fluorophore in the localized electromagnetic field. The quenching and enhancement of luminescence intensity of the dye molecules can also be explained as the transfer of electrons from dye to the silver nanoparticles and to an extent it can be attributed to the aggregation of dye molecules upon addition of silver nanoparticles.     

Photoreversible Fluorescent Modulation of Nanoparticles via One‐Step Miniemulsion Polymerization

A nitrobenzoxadiazolyl(NBD)‐based fluorescent dye and a photochromic spiropyran derivative are incorporated into polymeric nanoparticles via a one‐step miniemulsion polymerization. The diameter of the nanoparticles can be varied from approximately 40 nm to 80 nm by adjusting the polymerization conditions. The prepared nanoparticles exhibit the spectral properties of both NBD dye and spiropyran, indicating that the two chromophores are incorporated into the nanoparticles. The determined amount of NBD and spiropyran in the nanoparticles are about ≈85–90% of the feed amount, while the determined weight ratios of spiropyran to NBD in nanoparticles are very close to that of feed ratios, suggesting the miniemulsion polymerization is a suitable approach for incorporating multiple chromophores into individual nanoparticles with controlled amounts (content) and ratio. UV and visible light can be applied to modulate the fluorescence emission of NBD dye in nanoparticles. Upon UV irradiation, the spiropyran moieties in nanoparticles are converted to the open‐ring (McH form) structure and upon visible‐light irradiation they return to the closed‐ring (SP form) structure; as a result, the fluorescence of NBD can be reversibly “switched off” and “switched on”. Fluorescence resonance energy transfer from the excited NBD dye molecules to the McH form of the spiropyran moieties is the drives the fluorescence modulation. The nanoparticles display fairly good photoreversibility, photostability, and relatively fast photoresponsivity upon alternate UV/Vis irradiation. This class of photoresponsive nanoparticles may find applications in biological fields, such as labeling and imaging, as well as in optical fields, for example, individually light‐addressable nanoscale devices.     

Plasmonic enhancement of molecular fluorescence

Metallic nanoparticles are known to dramatically modify the spontaneous emission of nearby fluorescent molecules and materials. Here we examine the role of the nanoparticle plasmon resonance energy and nanoparticle scattering cross section on the fluorescence enhancement of adjacent indocyanine green (ICG) dye molecules. We find that enhancement of the molecular fluorescence by more than a factor of 50 can be achieved for ICG next to a nanoparticle with a large scattering cross section and a plasmon resonance frequency corresponding to the emission frequency of the molecule.     

Dual-emission fluorescent silica nanoparticle-based probe for ultrasensitive detection of Cu2+

An effective dual-emission fluorescent silica nanoparticle-based probe has been constructed for rapid and ultrasensitive detection of Cu(2+). In this nanoprobe, a dye-doped silica core served as a reference signal, thus providing a built-in correction for environmental effects. A response dye was covalently grafted on the surface of the silica nanoparticles through a chelating reagent for Cu(2+). The fluorescence of the response dye could be selectively quenched in the presence of Cu(2+), accompanied by a visual orange-to-green color switch of the nanoprobe. The nanoprobe provided an effective platform for reliable detection of Cu(2+) with a detection limit as low as 10 nM, which is nearly 2 × 10(3) times lower than the maximum level (～20 μM) of Cu(2+) in drinking water permitted by the U.S. Environmental Protection Agency (EPA). The high sensitivity was attributed to the strong chelation of Cu(2+) with polyethyleneimine (PEI) and a signal amplification effect. The nanoprobe constructed by this method was very stable, enabling the rapid detection of Cu(2+) in real water samples. Good linear correlations were obtained over the concentration range from 1 × 10(-7) to 8 × 10(-7) (R(2) = 0.99) with recoveries of 103.8-99.14% and 95.5-95.14% for industrial wastewater and lake water, respectively. Additionally, the long-wavelength emission of the response dye can avoid the interference of the autofluorescence of the biosystems, which facilitated their applications in monitoring Cu(2+) in cells. Furthermore, the nanoprobe showed a good reversibility; the fluorescence can be switched "off" and "on" by an addition of Cu(2+) and EDTA, respectively.     

A novel luminescent property from composite of ZnS nanocrystallites and organic chromophore molecules

ZnS nanocrystallites doped with organic dye (molecular chromophore) have been synthesized using a simple chemical method. Composite ZnS nanoparticles having different sizes and morphology can be obtained. The size of these composite nanoparticles is typically 10–30 nm. A dramatic increase in the luminescence intensity and a change in the emission wavelength have been observed from the composite nanoparticles. Because of the different structure and properties of dyes, the absorption, excitation and emission spectra of the composite nanoparticles vary with different dyes.     

Magnetic and fluorescent core-shell nanoparticles for ratiometric pH sensing

This paper describes the preparation of nanoparticles composed of a magnetic core surrounded by two successive silica shells embedding two fluorophores, showing uniform nanoparticle size (50-60 nm in diameter) and shape, which allow ratiometric pH measurements in the pH range 5-8. Uncoated iron oxide magnetic nanoparticles (～10 nm in diameter) were formed by the coprecipitation reaction of ferrous and ferric salts. Then, they were added to a water-in-oil microemulsion where the hydrophilic silica shells were obtained through hydrolysis and condensation of tetraethoxyorthosilicate together with the corresponding silylated dye derivatives-a sulforhodamine was embedded in the inner silica shell and used as the reference dye while a pH-sensitive fluorescein was incorporated in the outer shell as the pH indicator. The magnetic nanoparticles were characterized using vibrating sample magnetometry, dynamic light scattering, transmission electron microscopy, x-ray diffraction and Fourier transform infrared spectroscopy. The relationship between the analytical parameter, that is, the ratio of fluorescence between the sensing and reference dyes versus the pH was adjusted to a sigmoidal fit using a Boltzmann type equation giving an apparent pK(a) value of 6.8. The fluorescence intensity of the reference dye did not change significantly (～3.0%) on modifying the pH of the nanoparticle dispersion. Finally, the proposed method was statistically validated against a reference procedure using samples of water and physiological buffer with 2% of horse serum, indicating that there are no significant statistical differences at a 95% confidence level.     

Sensitive and selective detection of adenine using fluorescent ZnS nanoparticles

We have used fluorescent ZnS nanoparticles as a probe for the determination of adenine. A typical 2 × 10(-7) M concentration of adenine quenches 39.3% of the ZnS fluorescence. The decrease in ZnS fluorescence as a function of adenine concentration was found to be linear in the concentration range 5 × 10(-9)-2 × 10(-7) M. The limit of detection (LOD) of adenine by this method is 3 nM. Among the DNA bases, only adenine quenched the fluorescence of ZnS nanoparticles in the submicromolar concentration range, thus adding selectivity to the method. The amino group of adenine was important in determining the quenching efficiency. Steady-state fluorescence experiments suggest that one molecule of adenine is sufficient to quench the emission arising from a cluster of ZnS consisting of about 20 molecules. Time-resolved fluorescence measurements indicate that the adenine molecules block the sites on the surface of ZnS responsible for emission with the longest lifetime component. This method may be applied for the determination of adenine in biological samples since the measurements have been carried out at pH 7.     

Fluorescent Polymer Nanoparticles Based on Dyes: Seeking Brighter Tools for Bioimaging

Speed, resolution and sensitivity of today's fluorescence bioimaging can be drastically improved by fluorescent nanoparticles (NPs) that are many‐fold brighter than organic dyes and fluorescent proteins. While the field is currently dominated by inorganic NPs, notably quantum dots (QDs), fluorescent polymer NPs encapsulating large quantities of dyes (dye‐loaded NPs) have emerged recently as an attractive alternative. These new nanomaterials, inspired from the fields of polymeric drug delivery vehicles and advanced fluorophores, can combine superior brightness with biodegradability and low toxicity. Here, we describe the strategies for synthesis of dye‐loaded polymer NPs by emulsion polymerization and assembly of pre‐formed polymers. Superior brightness requires strong dye loading without aggregation‐caused quenching (ACQ). Only recently several strategies of dye design were proposed to overcome ACQ in polymer NPs: aggregation induced emission (AIE), dye modification with bulky side groups and use of bulky hydrophobic counterions. The resulting NPs now surpass the brightness of QDs by ≈10‐fold for a comparable size, and have started reaching the level of the brightest conjugated polymer NPs. Other properties, notably photostability, color, blinking, as well as particle size and surface chemistry are also systematically analyzed. Finally, major and emerging applications of dye‐loaded NPs for in vitro and in vivo imaging are reviewed.     

On the quenching of semiconductor quantum dot photoluminescence by proximal gold nanoparticles

Luminescent quantum dots (QDs) were proven to be very effective fluorescence resonance energy transfer donors with an array of organic dye acceptors, and several fluorescence resonance energy transfer based biosensing assemblies utilizing QDs have been demonstrated in the past few years. Conversely, gold nanoparticles (Au-NPs) are known for their capacity to induce strong fluorescence quenching of conventional dye donors. Using a rigid variable-length polypeptide as a bifunctional biological linker, we monitor the photoluminescence quenching of CdSe-ZnS QDs by Au-NP acceptors arrayed around the QD surface, where the center-to-center separation distance was varied over a broad range of values (approximately 50-200 Angstrom). We measure the Au-NP-induced quenching rates for such QD conjugates using steady-state and time-resolved fluorescence measurements and examine the results within the context of theoretical treatments based on the F?rster dipole-dipole resonance energy transfer, dipole-metal particle energy transfer, and nanosurface energy transfer. Our results indicate that nonradiative quenching of the QD emission by proximal Au-NPs is due to long-distance dipole-metal interactions that extend significantly beyond the classical F?rster range, in agreement with previous studies using organic dye-Au-NP donor-acceptor pairs.     

Investigation of the enhancement fluorescence of ethanol doped SiO2 nanoparticles

As one of the most commonly used solvents, ethanol exhibits weak fluorescence when excited by ultraviolet (UV) light. Until now, the fluorescence of ethanol-doped nanoparticles has not been studied. In this paper, eleven different concentrations of SiO2 nanoparticles (diameter 100 nm) were doped in ethanol, and corresponding colloids were formed. The excitation and emission spectra of the colloids were measured. The experimental results indicated that the SiO2 nanoparticles obviously enhanced the fluorescence of ethanol. Under excitation at 306 nm, the enhancement effect is the best when the concentration of SiO2 nanoparticles is 4.452 x 10(12) ml(-1), and the enhancement factor is nearly 50 times at the peak position of 360 nm. At the excitation wavelength of 360 nm, the enhancement effect is the best when the concentration of SiO2 nanoparticles is 1.113 x 10(13) ml(-1), and the enhancement factor is nearly 40 times at the peak position of 397 nm. The result of this article will reduce the test limit of ethanol by two magnitudes.     

Dipolar versus octupolar triphenylamine-based fluorescent organic nanoparticles as brilliant one- and two-photon emitters for (bio)imaging

Two related triphenylamine-based dipolar and octupolar fluorophores are used to prepare aqueous suspensions of fluorescent organic nanoparticles (FONs) via the reprecipitation method. The obtained spherical nanoparticles (30-40 nm in diameter) are fluorescent in aqueous solution (up to 15% fluorescence quantum yield) and exhibit extremely high one- and two-photon brightness, superior to those obtained for quantum dots. Despite the two chromophores showing similar fluorescence in solution, the fluorescence of FONs made from the octupolar derivative is significantly red-shifted compared to that generated by the dipolar FONs. In addition, the maximum two-photon absorption cross section of the FONs made from the octupolar derivative is 55% larger than that of the dipolar derivative FONs. The experimental observations provide evidence that the different molecular shape (rodlike versus three-branched) and charge distribution (dipolar versus octupolar) of the two chromophores strongly affect the packing inside the nanoparticles as well as their spectroscopic properties and colloidal stability in pure water. The use of these FONs as probes for biphotonic in-vivo imaging is investigated on Xenopus laevis tadpoles to test their utilization for angiography. When using FONs made from the octupolar dye, the formation of microagglomerates (2-5 μm scale) is observed in vivo, with subsequent lethal occlusion of the blood vessels. Conversely, the nanoparticles of the dipolar dye allow acute imaging of blood vessels thanks to their suitable size and brightness, while no toxic effect is observed. Such a goal cannot be achieved with the dissolved dye, which permeates the vessel walls.     

Optical detection and charge-state analysis of MALDI-generated particles with molecular masses larger than 5 MDa

Charged polystyrene nanoparticles are generated by matrix-assisted laser desorption/ionization (MALDI) and detected by laser-induced fluorescence (LIF) in a quadrupole ion trap. Employing the LIF technique, observations of individual fluorescent nanospheres (27 nm in diameter and containing 180 fluorescein dye equivalents) have been achieved with an average signal-to-noise ratio of approximately 10. With the trap operating at a frequency around 5 kHz, charge state analysis of the particles reveals that the number of charges carried by the spheres is between 1 and 10. It suggests a mass-to-charge ratio (m/z) in the range of 10(5)-10(6) for the MALDI-generated particles. To effectively trap such large particles (m > 5 MDa), damping of the particles' motions by using approximately 50 mTorr He buffer gas is absolutely required. Similar findings are obtained for particles with a nominal size of 1 microm in diameter, demonstrating that production of charged particles with a molecular mass as high as 10(12) Da is possible using the MALDI technique.     

Highly sensitive interfacial fluorescence sensing of deoxyribonucleic acid based on self-assembled multilayers

Fluorescent dye Acridine Orange (AO) was indirectly assembled onto the surface of colloidal gold nanoparticles modified quartz and gold wafer, with cysteine as mediation. Strong fluorescence emission was observed from AO self-assembled multilayers. Formation of the self-assembled multilayers was confirmed and primarily characterized by electrochemical and fluorescence methods. The effect of assembling substrate on fluorescence intensity of the film was discussed. The self-assembled multilayers offer a high sensitive interfacial fluorescence sensing of deoxyribonucleic acid with the detection limit of 7.145 × 10^− 9 g L^− 1 (3 S.D./K), which was five orders of magnitude lower than that of the bulk solution-phase. The self-assembled multilayers film has higher stability and good regeneration capacity.     

Fluorophore-gold nanoparticle complex for sensitive optical biosensing and imaging

Fluorophores have been extensively used as the signal mediator in biosensing and bioimaging for a long time. Enhancement of fluorescence can amplify the signal, thus improving the sensitivity, enabling earlier and accurate disease detection and diagnosis. Some metal nanoparticles, such as gold and silver, can generate a strong electromagnetic field on their surface (surface plasmon field) upon receiving photonic energy. When a fluorophore is placed in the field, the field can affect the fluorophore electrons participating in fluorescence emission and change the fluorescence output. The change can be from complete quenching to significant enhancement, depending on the metal type, particle size and shape, excitation/emission wavelengths and quantum yield of the fluorophore, and the distance between the fluorophore and the particle surface. In this study, the effects of these parameters on the fluorescence enhancement of commonly used fluorophores by gold nanoparticles (GNPs) are theoretically analyzed. Experimentally, an NIR contrast agent with enhanced fluorescence was developed by carefully tailoring the distance between Cypate (ICG based fluorophore) and a GNP, via biocompatible spacer constructs. The effect of the GNP size (3.7-16.4 nm) and spacer length (3.2-4.6 nm) on fluorescence enhancement was studied, and the spacer length that provided the significant enhancement was determined. The spacer of 3.9 nm with 16.4 nm GNP provided the fluorescence of 360% of the control. The experimental data qualitatively agreed with the theoretical results and, thus, the theoretical analysis can be used as a guide for significantly improving the sensitivity of existing fluorescent contrast agents by properly utilizing GNPs and spacers.     

Novel functionalized ternary copolymer fluorescent nanoparticles: synthesis, fluorescent characteristics and protein immobilization

Novel fluorescent poly(styrene-acrylamide-acrylic acid) nanoparticles (FPSAAN) were synthesized by means of soapless emulsion polymerization and being modified with hydrazine hydrate by hydrazinolysis. The azidocarbonyl groups which can be rapidly coupled with proteins under mild condition were introduced onto the fluorescent nanoparticles by azido-reaction. Bovine serum albumin (BSA) was selected as a model protein to be covalently immobilized on the azidocarbonyl FPSAAN. Atom force microscopy (AFM), Transmission electron microscopy (TEM), Ultraviolet-visible (UV/Vis) spectrometer, Fourier transforms infrared spectrometer (FTIR), nanoparticle size analyzer and fluorescence spectrophotometer were used to characterize the FPSAAN. Results showed that FPSAAN had a regular spherical shape, and a dramatic narrow size distribution (polydispersity index 0.046 +/- 0.009). The fluorescence intensity of FPSAAN (lambda(ex)/lambda(em) = 253/306 nm), hydrazide-FPSAAN (lambda(ex/)lambda(em) = 260/326 nm), and protein-immobilized FPSAAN (lambda(ex)/lambda(em) = 258/325 nm) was linearly related to the concentration ranging from 1.0 x 10(-3) g l(-1) to 10.0 x 10(-3) g l(-1). The linear relationship was obtained. The equations are y = 52.808x + 16.465 (R (2) = 0.9927), y = 5.1814x + 4.1535 (R (2) = 0.9935) and y = 5.2227x + 5.2883 (R (2) = 0.9937), respectively. In addition, external factors such as pH and ionic strength exert a slight influence on fluorescent properties. The experiments of the immobilization of BSA indicated that FPSAAN with azidocarbonyl groups could be covalently coupled with BSA at the rate of 41.1%. Meanwhile, hCG antibody immobilized FPSAAN have the similar fluorescence characteristics to BSA immobilized FPSAAN. Only negligible difference of the fluorescence characteristics can be found. Furthermore, the fluorescence characteristics of hCG antibody immobilized FPSAAN have not been obviously affected after mixed with the hCG antigen and human plasma. These novel azidocarbonyl FPSAAN with stable fluorescence and active functional azidocarbonyl groups could be used as a promising fluorescent probe for quantitative detection, protein immobilization, cell labeling research and early rapid clinical diagnostics.     

Synthesis, characterization, and application of Eu(III), Tb(III), Sm(III), and Dy(III) lanthanide chelate nanoparticle labels

Preparation and characterization of europium(III), terbium(III), samarium(III), and dysprosium(III) polystyrene nanoparticle labels with lanthanide-specific fluorescence properties has been presented. Emulsion copolymerization of styrene and acrylic acid was used to synthesize uniform-sized nanoparticles approximately 45 nm in diameter. Europium(III) and samarium(III) lanthanides were chelated with 2-naphthoyltrifluoroacetone and trioctylphosphine oxide to dye the spherical particles, whereas terbium(III) and dysprosium(III) chelate complexes contained a newly synthesized ligand, 4-(2,4,6-tridecyloxyphenyl)pyridine-2,6-dicarboxylic acid. The fluorescence properties of the four lanthanides-including a wide Stokes shift, a narrow emission peak, and long fluorescence lifetime-were retained despite the incorporation into the nanoparticles. Furthermore, the nanoparticles, containing more than 1000 lanthanide chelates, were detectable at label concentrations 3 orders of magnitude lower than the corresponding soluble lanthanide chelate labels. The applicability of the labels prepared was demonstrated by a heterogeneous sandwich-type immunoassay for human prostate-specific antigen, where the lowest limits of detection of 1.6, 2.4, 10.1, and 114.2 ng/L were achieved using europium(III), terbium(III), samarium(III), and dysprosium(III) nanoparticles, respectively. The spectral and functional properties of the lanthanide-embedded polystyrene nanoparticles developed here suggest that the technology is applicable for high-sensitivity multicolor assays.