Ramachandram Badugu

Advances in Surface-Enhanced Fluorescence

We report recent achievements in metal-enhanced fluorescence from our laboratory. Several fluorophore systems have been studied on metal particle-coated surfaces and in colloid suspensions. In particular, we describe a distance dependent enhancement on silver island films (SIFs), release of self-quenching of fluorescence near silver particles, and the applications of fluorescence enhancement near metalized surfaces to bioassays. We discuss a number of methods for various shaped silver particle deposition on surfaces.

A Glucose Sensing Contact Lens: A Non-Invasive Technique for Continuous Physiological Glucose Monitoring

We have developed a range of glucose sensing contact lenses, using a daily, disposable contact lens embedded with newly developed boronic acid containing fluorophores. Our findings show that our approach may be suitable for the continuous monitoring of tear glucose levels in the range 50–1000 μM, which typically track blood glucose levels, which are ≈5–10 fold higher. Our non-invasive approach may well offer an alternative solution to current invasive glucose monitoring techniques for diabetes, such as “finger pricking.”

Ophthalmic Glucose Monitoring Using Disposable Contact Lenses—A Review

We have developed a range of disposable and colorless tear glucose sensing contact lenses, using off-the-shelf lenses embedded with new water soluble, highly fluorescent and glucose sensitive boronic acid containing fluorophores. The new lenses are readily able to track tear glucose levels and therefore blood glucose levels, which are ideally suited for potential use by diabetics. The fluorescence responses from the lenses can be monitored using simple excitation and emission detection devices. The novelty of our approach is two fold. Firstly, the notion of sensing extremely low glucose concentrations in tears, which track blood levels, by our contact lens approach, and secondly, the unique compatibility of our new glucose signaling probes with the internal mildly acidic contact lens environment. The new lenses are therefore ideal for the non-invasive and continuous monitoring of tear glucose, with about 15-min response time, and a measured shelf life in excess of 3 months. In this review article, we show that fluorescence based signaling using plastic disposable lenses, which have already been industrially optimized with regard to vision correction and oxygen/analyte permeability etc, may a notable alternative to invasive and random finger pricking, the most widely used glucose monitoring technology by diabetics.

Fluorescence sensors for monosaccharides based on the 6-methylquinolinium nucleus and boronic acid moiety: potential application to ophthalmic diagnostics

Continuous monitoring of glucose levels in human physiology is important for the long-term management of diabetes. New signaling methods/probes may provide an improved technology to monitor glucose and other physiologically important analytes. The glucose sensing probes, BMQBAs, fabricated using the 6-methylquinolinium moiety as a fluorescent indicator, and boronic acid as a chelating group, may have versatile applications in glucose sensing because of their unique properties. In this paper we discuss the design logic, synthesis, characterization and spectral properties of three new isomeric glucose sensors (BMQBAs), and a control compound (BMQ) in the presence and absence of sugars. The sensing ability of the new probes is based on a charge neutralization and stabilization mechanism upon sugar binding. The new probes have attractive fluorescence quantum yields, are highly water-soluble, and have spectral characteristics compatible with cheap and portable LEDs and LDs. One of the probes, o-BMQBA, has a sugar bound pK(a) of 6.1, and a dissociation constant K(D) of 100mM glucose. These probes have been designed specifically to respond to tear glucose in a contact lens polymer for ophthalmic glucose monitoring, where the reduced sugar bound pK(a) affords for sensing, in a lens environment that we have previously shown to be mildly acidic.

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.

Radiative decay engineering 7: Tamm state-coupled emission using a hybrid plasmonic–photonic structure

There is a continuing need to increase the brightness and photostability of fluorophores for use in biotechnology, medical diagnostics, and cell imaging. One approach developed during the past decade is to use metallic surfaces and nanostructures. It is now known that excited state fluorophores display interactions with surface plasmons, which can increase the radiative decay rates, modify the spatial distribution of emission, and result in directional emission. One important example is surface plasmon-coupled emission (SPCE). In this phenomenon, the fluorophores at close distances from a thin metal film, typically silver, display emission over a small range of angles into the substrate. A disadvantage of SPCE is that the emission occurs at large angles relative to the surface normal and at angles that are larger than the critical angle for the glass substrate. The large angles make it difficult to collect all of the coupled emission and have prevented the use of SPCE with high-throughput and/or array applications. In the current article, we describe a simple multilayer metal-dielectric structure that allows excitation with light that is perpendicular (normal) to the plane and provides emission within a narrow angular distribution that is normal to the plane. This structure consists of a thin silver film on top of a multilayer dielectric Bragg grating, with no nanoscale features except for the metal or dielectric layer thicknesses. Our structure is designed to support optical Tamm states, which are trapped electromagnetic modes between the metal film and the underlying Bragg grating. We used simulations with the transfer matrix method to understand the optical properties of Tamm states and localization of the modes or electric fields in the structure. Tamm states can exist with zero in-plane wavevector components and can be created without the use of a coupling prism. We show that fluorophores on top of the metal film can interact with the Tamm state under the metal film and display Tamm state-coupled emission (TSCE). In contrast to SPCE, the Tamm states can display either S or P polarization. The TSCE angle is highly sensitive to wavelength, which suggests the use of Tamm structures to provide both directional emission and wavelength dispersion. Metallic structures can modify fluorophore decay rates but also have high losses. Photonic crystals have low losses but may lack the enhanced light-induced fields near metals. The combination of plasmonic and photonic structures offers the opportunity for radiative decay engineering to design new formats for clinical testing and other fluorescence-based applications.

Wavelength-ratiometric probes for the selective detection of fluoride based on the 6-aminoquinolinium nucleus and boronic acid moiety.

Herein we report a set of new water-soluble fluorescent probes (N-boronobenzyl-6-aminoquinolinium bromides, BAQBAs) sensitive to aqueous fluoride. These probes shows spectral shifts and intensity changes in the presence of fluoride, in a wavelength ratiometric and colorimetric manner, enabling the detection of fluoride concentrations at visible wavelengths, in the concentration range ≈1-300 mM. Although the sensing mechanism is different for fluoride as compared to the other halides, we have tested the utility of these probes towards the other halides, and the results reveal that the BAQBAs are in fact potential candidates towards the sensing of the all the halides, but in different concentration ranges. As the probes are based on the boronic acid moiety, which is a well-known fluoride and sugar chelator group, we have investigated the response of sugars (such as glucose and fructose, which are present in biological fluids and foodstuffs) as interferences in fluoride detection using these probes. Interestingly, the BAQBAs show a suppressed sugar response potentially allowing for the predominant fluoride sensitivity. In addition to physiological sugars, we also have assessed the response of aqueous halides as potential interferents, or indeed analytes to be sensed, and show that the new boronic acid containing probes respond well to aqueous fluoride in the presence of a high background of other species, such as in a biological cocktail of 50 mM Glucose, 50 mM aqueous Chloride and 5 mM Fructose.

Wavelength-ratiometric near-physiological pH sensors based on 6-aminoquinolinium boronic acid probes

We describe the pH response of a set of isomeric water-soluble fluorescent probes based on both the 6-aminoquinolinium and boronic acid moieties. These probes show spectral shifts and intensity changes with pH, in a wavelength-ratiometric and colorimetric manner. Subsequently, changes in pH can readily be determined around the physiological level. Although boronic acid containing probes are known to exhibit pH sensitivity along with an ability for saccharide binding/chelating, the new probes reported here are considered to be unique and show an unperturbed pH response, even in the presence of high concentrations of background saccharide, such as with glucose and fructose, allowing for the predominant pH sensitivity. The response of the probes is based on the ability of the boronic acid group to interact with strong bases like OH(-), changing from the neutral form of the boronic acid group, R-B(OH)(2), to the anionic ester, R-B(-)(OH)(3), form, which is an electron donating group. The presence of an electron deficient quaternary heterocyclic nitrogen center and a strong electron donating amino group in the 6-position of the quinolinium backbone, provides for the spectral changes observed upon OH(-) complexation. In addition, by comparing the results obtained with systems separately incorporating 6-methoxy or 6-methyl substituents, the suppressed response towards monosaccharides, such as with glucose and fructose, can clearly be observed for these systems. Finally we compare our results to those of a control compound, BAQ, which does not contain the boronic acid group, allowing a rationale of the spectral changes to be made.

A glucose-sensing contact lens: a new approach to noninvasive continuous physiological glucose monitoring

We have developed a new technology for the non-invasive continuous monitoring of tear glucose using a daily use, disposable contact lens, embedded with sugar-sensing boronic acid containing fluorophores. Our findings show that our approach may be suitable for the continuous monitoring of tear glucose levels in the range 50 - 500 μM, which track blood glucose levels that are typically ≈ 5-10 fold higher. We initially tested the sensing concept with well-established, previously published, boronic acid probes and the results could conclude the used probes, with higher pKa values, are almost insensitive toward glucose within the contact lens, attributed to the low pH and polarity inside the lens. Subsequently, we have developed a range of probes based on the quinolinium backbone, having considerably lower pKa values, which enables them to be suitable to sense the physiological glucose in the acidic pH contact lens. Herein we describe the results based on our findings towards the development of glucose sensing contact lens and therefore an approach to non-invasive continuous monitoring of tear glucose using a contact lens.

Directing fluorescence with plasmonic and photonic structures.

Fluorescence technology pervades all areas of chemical and biological sciences. In recent years, it is being realized that traditional fluorescence can be enriched in many ways by harnessing the power of plasmonic or photonic structures that have remarkable abilities to mold the flow of optical energy. Conventional fluorescence is omnidirectional in nature, which makes it difficult to capture the entire emission. Suitably designed emission directivity can improve collection efficiency and is desirable for many fluorescence-based applications like sensing, imaging, single molecule spectroscopy, and optical communication. By incorporating fluorophores in plasmonic or photonic substrates, it is possible to tailor the optical environment surrounding the fluorophores and to modify the spatial distribution of emission. This promising approach works on the principle of near-field interaction of fluorescence with spectrally overlapping optical modes present in the substrates. In this Account, we present our studies on directional emission with different kinds of planar metallic, dielectric, and hybrid structures. In metal-dielectric substrates, the coupling of fluorescence with surface plasmons leads to directional surface-plasmon-coupled emission with characteristic dispersion and polarization properties. In one-dimensional photonic crystals (1DPC), fluorophores can interact with Bloch surface waves, giving rise to sharply directional Bloch surface wave-coupled emission. The interaction of fluorescence with Fabry-Pérot-like modes in metal-dielectric-metal substrates and with Tamm states in plasmonic-photonic hybrid substrates provides beaming emission normal to the substrate surface. These interesting features are explained in the context of reflectivity dispersion diagrams, which provide a complete picture of the mode profiles and the corresponding coupled emission patterns. Other than planar substrates, specially fabricated plasmonic nanoantennas also have tremendous potential in controlling and steering fluorescence beams. Some representative studies by other research groups with various nanoantenna structures are described. While there are complexities to near-field interactions of fluorescence with plasmonic and photonic structures, there are also many exciting possibilities. The routing of each emission wavelength along a specific direction with a given angular width and polarization will allow spatial and spectral multiplexing. Directional emission close to surface normal will be particularly useful for microscopy and array-based studies. Application-specific angular emission patterns can be obtained by varying the design parameters of the plasmonic/photonic substrates in a flexible manner. We anticipate that the ability to control the flow of emitted light in the nanoscale will lead to the development of a new generation of fluorescence-based assays, instrumentation, portable diagnostics, and emissive devices.