Mustafa H. Chowdhury

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.

Aluminum Nanoparticles as Substrates for Metal-Enhanced Fluorescence in the Ultraviolet for the Label-Free Detection of Biomolecules

We use finite-difference time-domain calculations to show that aluminum nanoparticles are efficient substrates for metal-enhanced fluorescence (MEF) in the ultraviolet (UV) for the label-free detection of biomolecules. The radiated power enhancement of the fluorophores in proximity to aluminum nanoparticles is strongly dependent on the nanoparticle size, fluorophore-nanoparticle spacing, and fluorophore orientation. Additionally, the enhancement is dramatically increased when the fluorophore is between two aluminum nanoparticles of a dimer. Finally, we present experimental evidence that functionalized forms of amino acids tryptophan and tyrosine exhibit MEF when spin-coated onto aluminum nanostructures.

Metal-enhanced chemiluminescence: Radiating plasmons generated from chemically induced electronic excited states

In this letter, we report the observation of metal-enhanced chemiluminescence. Silver Island films, in close proximity to chemiluminescence species, can significantly enhance luminescence intensities; a 20-fold increase in chemiluminescence intensity was observed as compared to an identical control sample containing no silver. This suggests the use of silver nanostructures in the chemiluminescence-based immunoassays used in the biosciences today, to improve signal and therefore analyte detectability. In addition, this finding suggests that surface plasmons can be directly excited by chemically induced electronically excited luminophores, a significant finding toward our understanding of fluorophore-metal interactions and the generation of surface plasmons.

Metal-Enhanced Chemiluminescence

In this short paper we report the interactions of silver island films with chemiluminescing species. Our findings show that silver island films can increase the detectability of chemiluminescent reactions/species, with an approximately 5-fold increase in signal intensity. This finding not only suggests the use of silver nanostructures to amplify chemiluminscent signatures in assay platforms, and therefore increase the detectability of analytes or biospecies, but more importantly, suggests that surface plasmons can be directly excited by chemically induced electronically excited molecules. This finding is of significance towards our understanding of fluorophore–metal interactions, a relatively new near-field fluorescence concept, recently named metal-enhanced fluorescence and also radiative decay engineering.

Use of surface-enhanced Raman spectroscopy for the detection of human integrins.

Current research has revealed the importance of a class of cell surface proteins called integrins in various vital physiological functions such as blood clotting, regulation of blood pressure, tissue blood flow, and vascular remodeling. The key to integrin functionality is its ability to mediate force transmission by interacting with the extracellular matrix and cytoskeleton. In addition, they play a role in signal transduction via their connection with the proteins in focal adhesion (FA) points. To understand the complex mechanism of cell-cell and cell-extracellular matrix (ECM) adhesion that is responsible for these diverse biochemical interactions, it is necessary to identify the integrins on cells and monitor their interaction with various ligands. To this end, for the first time, we employ surface-enhanced Raman spectroscopy (SERS) to detect integrins. The results show the capability using SERS to detect the integrins to the nanomolar concentration regime and to distinguish between two different kinds of integrins, alphaVbeta3 and alpha5beta1, that are present in vascular smooth muscle cells (VSMCs). It is anticipated that the SERS approach will potentially help elucidate the mechanism of integrin-ligand interactions in a variety of phenomena of physiological importance.

Plasmon-Controlled Fluorescence Towards High-Sensitivity Optical Sensing

Fluorescence spectroscopy is widely used in chemical and biological research. Until recently most of the fluorescence experiments have been performed in the far-field regime. By far-field we imply at least several wavelengths from the fluorescent probe molecule. In recent years there has been 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 dramatically be 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. Fluorophores in the excited state can create plasmons that radiate into the far-field and fluorophores in the ground state can interact with and be excited by surface plasmons. These reciprocal 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 these phenomena as plasmon-controlled fluorescence (PCF). An overview of the recent work on metal-fluorophore interactions is presented. Recent research combining plasmonics and fluorescence suggest that PCF could lead to new classes of experimental procedures, novel probes, bioassays, and devices.

Imaging three-dimensional light propagation through periodic nanohole arrays using scanning aperture microscopy.

The authors introduce a technique for three-dimensional (3D) imaging of the light transmitted through periodic nanoapertures using a scanning probe to perform optical sectioning microscopy. For a 4×4 nanohole array, the transmitted light displays intensity modulations along the propagation axis, with the maximum intensity occurring at 450 μm above the surface. The propagating fields show low divergence, suggesting a beaming effect induced by the array. At distances within 25 μm from the surface, they observe subwavelength confinement of light propagating from the individual nanoholes. Hence, this technique can potentially be used to map the 3D distribution of propagating light, with high spatial resolution.

Single-Molecule Spectroscopic Study of Enhanced Intrinsic Phycoerythrin Fluorescence on Silver Nanostructured Surfaces

In this paper, we report on steady-state and time-resolved single-molecule fluorescence measurements performed on a phycobiliprotein, R-phycoerythrin (RPE), assembled on silver nanostructures. Single-molecule measurements clearly show that RPE molecules display a 10-fold increase in fluorescence intensity, with a 7-fold decrease in lifetime when they are assembled on silver nanostructured surfaces, as compared to control glass slides. The emission spectrum of individual RPE molecules also displays a significant fluorescence enhancement on silver nanostructures as compared to glass. From intensity and lifetime histograms, it is clear that the intensities as well as lifetimes of individual RPE molecules on silver nanostructures are more heterogeneously distributed than that on glass. This single-molecule study provides further insight on the heterogeneity in the fluorescence intensity and lifetimes of the RPE molecules on both glass and SiFs surfaces, which is otherwise not possible to observe using ensemble measurements. Finite-difference time-domain calculations have been performed to study the enhanced near-fields induced around silver nanoparticles by a radiating excited-state fluorophore, and the effect of such enhanced fields on the fluorescence enhancement observed is discussed.

Detection of differences in oligonucleotide-influenced aggregation of colloidal gold nanoparticles using absorption spectroscopy

A rapid, simple, and reproducible assay is described that can be used to detect differences in the ability of oligonucleotides to influence the aggregation of colloidal gold nanoparticles. The aggregation reaction of the gold colloid was monitored through UV-visible absorption spectroscopy. Single isolated colloidal gold particles have a surface plasmon resonance manifested as a single absorbance peak at approximately 520 nm, and aggregated gold complexes develop new red-shifted peaks/shoulders depending on the nature and extent of the aggregated complex. A simple ratiometric study of the area under the single and aggregated plasmon resonance peaks thus gives information about the extent of the aggregation. It is postulated that differences in dynamic flexibility of the oligonucleotides affect their influence on the aggregation state of the gold nanoparticles. The results of this study provide new clues toward unraveling the causes behind the preferential affinity of the Hermes transposable element for certain insertion sites compared to other sequences that also contain recognizable target sites. The technique is robust and thus can potentially be used to study similar questions for numerous transposable elements and target sequences.

Effect of Nanohole Spacing on the Self-Imaging Phenomenon Created by the Three-Dimensional Propagation of Light through Periodic Nanohole Arrays.

We present a detailed study of the inter-nanohole distance that governs the self-imaging phenomenon created by the three-dimensional propagation of light through periodic nanohole arrays on plasmonic substrates. We used scanning near-field optical microscopy (SNOM) to map the light intensity distributions at various heights above 10×10 nanohole arrays of varying pitch sizes on silver films. Our results suggest the inter-hole spacing has to be greater than the wavelength of the incident light to create the self-imaging phenomenon. We also present Finite-Difference Time-Domain (FDTD) calculations which show qualitative corroboration of our experimental results. Both our experimental and FDTD results show that the self-imaging phenomenon is more pronounced for structures with larger pitch sizes. We believe this self-imaging phenomenon is related to the Talbot imaging effect that has also been modified by a plasmonic component and can potentially be used to provide the basis for a new class of optical microscopes.