Enhanced single molecule fluorescence and reduced observation volumes on nanoporous gold (NPG) films.

We report single molecule fluorescence studies on nanoporous gold films. We observed dramatically enhanced intensities in individual immobilized fluorophores and reduced effective observation volumes in the nanopores. The enhanced localized plasmon field existing in the "nanovoid" regions can be visualized from the overlaid mapping image derived from surface reflectance and fluorescence emission.

Tuning Fluorescence Direction with Plasmonic Metal-Dielectric- Metal Substrates.

Controlling the emission properties of fluorophores is essential for improving the performance of fluorescence-based techniques in modern biochemical research, medical diagnosis, and sensing. Fluorescence emission is isotropic in nature, which makes it difficult to capture more than a small fraction of the total emission. Metal- dielectric-metal (MDM) substrates, discussed in this Letter, convert isotropic fluorescence into beaming emission normal to the substrate. This improves fluorescence collection efficiency and also opens up new avenues for a wide range of fluorescence-based applications. We suggest that MDM substrates can be readily adapted for multiple uses, such as in microarray formats, for directional fluorescence studies of multiple probes or for molecule-specific sensing with a high degree of spatial control over the fluorescence emission. SECTION: Physical Processes in Nanomaterials and Nanostructures.

Radiative decay engineering 6: fluorescence on one-dimensional photonic crystals.

During the past decade the interactions of fluorophores with metallic particles and surfaces has become an active area of research. These near-field interactions of fluorophores with surface plasmons have resulted in increased brightness and directional emission. However, using metals has some disadvantages such as quenching at short fluorophore-metal distances and increased rates of energy dissipation due to lossy metals. These unfavorable effects are not expected in dielectrics. In this article, we describe the interactions of fluorophores with one-dimensional (1D) photonic crystals (PCs), which have alternating layers of dielectrics with dimensions that create a photonic band gap (PBG). Freely propagating light at the PBG wavelength will be reflected. However, similar to metals, we show that fluorophores within near-field distances of the 1DPC interacts with the structure. Our results demonstrate that these fluorophores can interact with both internal modes and Bloch surface waves (BSWs) of the 1DPC. For fluorophores on the surface of the 1DPC, the emission dominantly occurs through the 1DPC and into the substrate. We refer to these two phenomena together as Bragg grating-coupled emission (BGCE). Here we describe our preliminary results on BGCE. 1DPCs are simple to fabricate and can be handled and reused without damage. We believe that BGCE provides opportunities for new formats for fluorescence detection and sensing.

Direct observation to chemokine receptor 5 on T-lymphocyte cell surface using fluorescent metal nanoprobes.

Chemokine receptor 5 (CCR5) is a cell surface protein required for HIV-1 infection. It is important to detect the amount and observe the spatial distribution of the CCR5 receptors on the cell surfaces. In this report, we describes the metal nanoparticles which were specially designed as molecular fluorescent probes for imaging of CCR5 receptors on the T-lymphocytic PM1 cell surfaces. These CCR5 monoclonal antibodies (mAbs) metal complexes were prepared by labeling mAbs with Alexa Fluor 680 followed by covalent binding the labeled mAbs on the 20 nm silver nanoparticles. Compared with the labeled mAbs without metal, the mAb-metal complexes were found to display enhanced emission intensity and shortened lifetime due to interactions between fluorophores and metal. The mAb-metal complexes were incubated with the PM1 cell lines. The confocal fluorescent intensity and lifetime cell images were recorded on single cells. It was observed that the mAb-metal complexes could be clearly distinguished from the cellular autofluorescence. By analyzing a pool of cell images, we observed that most CCR5 receptors appeared as clusters on the cell surfaces. The fluorophore-metal complexes developed in this report are generally useful for detection of cell surface receptors and provide a new class of probe to study the interaction between the CCR5 receptors with viral gp120 during HIV infections.

Plasmon-controlled fluorescence: a new paradigm in fluorescence spectroscopy.

Fluorescence spectroscopy is widely used in biological research. Until recently, essentially all fluorescence experiments were performed using optical energy which has radiated to the far-field. By far-field we mean at least several wavelengths from the fluorophore, but propagating far-field radiation is usually detected at larger macroscopic distances from the sample. In recent years there has been a growing interest in the interactions of fluorophores with metallic surfaces or particles. Near-field interactions are those occurring within a wavelength distance of an excited fluorophore. The spectral properties of fluorophores can be dramatically altered by near-field interactions with the electron clouds present in metals. These interactions modify the emission in ways not seen in classical fluorescence experiments. In this review we provide an intuitive description of the complex physics of plasmons and near-field interactions. Additionally, we summarize the recent work on metal-fluorophore interactions and suggest how these effects will result in new classes of experimental procedures, novel probes, bioassays and devices.

Single molecule photophysics near metallic nanostructures.

Metal-enhanced fluorescence (MEF) is useful in single molecule detection (SMD) by increasing the photostability, brightness and increase in radiative decay rates of fluorophores. We have investigated MEF from an individual fluorophore tethered to a single silver nanoparticle and also a single fluorophore between a silver dimer. The fluorescence lifetime results revealed a near-field interaction mechanism of fluorophore with the metal particle. Finite-difference time-domain (FDTD) calculations were employed to study the distribution of electric field near the metal monomer and dimer. The coupling effect of metal particles on the fluorescence enhancement was studied. We have also investigated the photophysics of FRET near metal nanoparticles and our preliminary results suggest an enhanced FRET efficiency in the presence of a metal nanoparticle. In total, our results demonstrate improved detectability at the single molecule level for a variety of fluorophores and quantum dots in proximity to the silver nanoparticles due to the near-field metal-fluorophore interactions.

Plasmon-controlled fluorescence: A new detection technology.

Fluorescence is widely used in biological research. Future advances in biology and medicine often depend on the advances in the capabilities of fluorescence measurements. In this overview paper we describe how a combination of fluorescence, and plasmonics, and nanofabrication can fundamentally change and increase the capabilities of fluorescence technology. This change will be based on the use of surface plasmons which are collective oscillations of free electrons in metallic surfaces and particles. Surface plasmon resonance is now used to measure bioaffinity reactions. However, the uses of surface plasmons in biology are not limited to their optical absorption or extinction. We have shown that fluorophores in the excited state can create plasmons which radiate into the far field; additionally fluorophores in the ground state can interact with and be excited by surface plasmons. These interactions suggest that the novel optical absorption and scattering properties of metallic nanostructures can be used to control the decay rates, location and direction of fluorophore emission. We refer to this technology as plasmon-controlled fluorescence. We predict that plasmon-controlled fluorescence (PCF) will result in a new generation of probes and devices. PCF is likely to allow design of structures which enhance emission at specific wavelengths and the creation of new devices which control and transport the energy from excited fluorophores in the form of plasmons, and then convert the plasmons back to light.

Waveguide-modulated surface plasmon-coupled emission of Nile blue in poly(vinyl alcohol) thin films.

Surface plasmon-coupled emission (SPCE) phenomenon is the coupling of excited fluorophores near a silver film with surface plasmons, resulting in directional emission into the underlying glass substrates. We report a complex coupling of Nile Blue fluorophore with 50 nm silver mirror, resulting in emission at several angles in the glass substrate, with either s or p polarization. This complex pattern of directional and polarized emission appears to be due to optical waveguide effects occurring when the sample thickness becomes comparable to the emission wavelength. We expect waveguide-modulated SPCE to have applications to biophysics and sensing.

Ultraviolet surface plasmon-coupled emission using thin aluminum films.

Surface plasmon-coupled emission (SPCE) is the directional radiation of light into a substrate due to excited fluorophores above a thin metal film. To date, SPCE has only been observed with visible wavelengths using silver or gold films. We now show that SPCE can be observed in the ultraviolet region of the spectrum using thin (20 nm) aluminum films. We observed directional emission in a quartz substrate from the DNA base analogue 2-aminopurine (2-AP). The SPCE radiation occurs within a narrow angle at 59 degrees from the normal to the hemicylindrical prism. The excitation conditions precluded the creation of surface plasmons by the incident light. The directional emission at 59 degrees is almost completely p-polarized, consistent with its origin from surface plasmons due to coupling of excited 2-AP with the aluminum. The emission spectra and lifetimes of the SPCE are those characteristic of 2-AP. Different emission wavelengths radiate at slightly different angles on the prism providing intrinsic spectral resolution from the aluminum film. These results indicate that SPCE can be used with numerous UV-absorbing fluorophores, suggesting biochemical applications with simultaneous surface plasmon resonance and SPCE binding assays.

Effects of Sample Thickness on the Optical Properties of Surface Plasmon-Coupled Emission.

In recent reports, we demonstrated coupling of excited fluorophores with thin silver or gold films resulting in directional surface plasmon-coupled emission (SPCE) through the silver film and into the glass substrate. In the present report, we describe the spectral and spatial properties of SPCE from sulforhomamine 101 in polyvinyl alcohol (PVA) films of various thicknesses on 50-nm silver films. The PVA thickness varied from about 30 to 750 nm. In thin PVA films with a thickness less than 160 nm, SPCE occurred at a single angle in the glass substrate and displayed only p polarization. As the PVA thickness increased to 300 nm, we observed SPCE at two angles, with different s or p polarization for each angle. For PVA films from 500 to 750 nm thick, we observed SPCE at three or four angles, with alternating s and p polarizations. The multiple rings of SPCE and the unusual s-polarized emission are consistent with the expected waveguide modes in the silver-PVA composite film. However, in contrast to our expectations, the average lifetimes of SPCE were not substantially changed from the PVA films. The observation of SPCE at multiple angles and with different polarization opens new opportunities for the use of SPCE to study anisotropic systems or to develop unique sensing devices.

Surface plasmon-coupled directional fluorescence emission.

Directional fluorescence emission of a sulforhodamine 101 in polyvinyl alcohol film has been observed from samples deposited on semi-transparent silver mirror. The fully p-polarized fluorescence emerges through the glass prism in form of hollow cone. The angle of this cone of emission depends on the thickness of the sample, and does not depend on the mode of excitation. The angular dependence of surface plasmon-coupled emission (SPCE) on the sample thickness has been discussed as well as its relevance to the surface plasmon resonance (SPR) analysis.