Microbubble resonators for scattering-free absorption spectroscopy of nanoparticles.

We present a proof-of-concept experiment where the absorbance spectra of suspensions of plasmonic nanoparticles are accurately reconstructed through the photothermal conversion that they mediate in a microbubble resonator. This thermal detection produces spectra that are insensitive towards light scattering in the sample, as proved experimentally by comparing the spectra of acqueos gold nanorods suspensions in the presence or absence of milk powder. In addition, the microbubble system allows for the interrogation of small samples (below 40 nl) while using a low-intensity beam (around 20 µW) for their excitation. In perspective, this system could be implemented for the characterization of turbid biological fluids through their optical absorption, especially when considering that the microbubble resonator naturally interfaces to a microfluidic circuit and may easily fit within portable or on-chip devices.

Microbubble Resonators for All-Optical Photoacoustics of Flowing Contrast Agents.

In this paper, we implement a Whispering Gallery mode microbubble resonator (MBR) as an optical transducer to detect the photoacoustic (PA) signal generated by plasmonic nanoparticles. We simulate a flow cytometry experiment by letting the nanoparticles run through the MBR during measurements and we estimate PA intensity by a Fourier analysis of the read-out signal. This method exploits the peaks associated with the MBR mechanical eigenmodes, allowing the PA response of the nanoparticles to be decoupled from the noise associated with the particle flow whilst also increasing the signal-to-noise ratio. The photostability curve of a known contrast agent is correctly reconstructed, validating the proposed analysis and proving quantitative PA detection. The experiment was run to demonstrate the feasible implementation of the MBR system in a flow cytometry application (e.g., the detection of venous thrombi or circulating tumor cells), particularly regarding wearable appliances. Indeed, these devices could also benefit from other MBR features, such as the extreme compactness, the direct implementation in a microfluidic circuit, and the absence of impedance-matching material.

Selective coupling of Whispering Gallery Modes in film coated micro-resonators.

Whispering Gallery Mode (WGM) micro-resonators like microspheres or microtoroids are typically used as high-Q cavity substrate on which a functional film coating is deposited. In order to exploit the coating properties a critical step is the efficient excitation of WGMs mainly contained inside the deposited layer. We developed a simple method able to assess whether or not these modes are selectively excited. The method is based on monitoring the thermal shift of the excited resonance, which uniquely depends on the thermo-optic coefficient and on the thermal expansion coefficient of the material in which the mode is embedded.

THz Pyro-Optical Detector Based on LiNbO3 Whispering Gallery Mode Microdisc Resonator.

This study analyzes the capabilities of a LiNbO3 whispering gallery mode microdisc resonator as a potential bolometer detector in the THz range. The resonator is theoretically characterized in the stationary regime by its thermo-optic and thermal coefficients. Considering a Q-factor of 107, a minimum detectable power of 20 µW was evaluated, three orders of magnitude above its noise equivalent power. This value opens up the feasibility of exploiting LiNbO3 disc resonators as sensitive room-temperature detectors in the THz range.

Biosensing by WGM Microspherical Resonators.

Whispering gallery mode (WGM) microresonators, thanks to their unique properties, have allowed researchers to achieve important results in both fundamental research and engineering applications. Among the various geometries, microspheres are the simplest 3D WGM resonators; the total optical loss in such resonators can be extremely low, and the resulting extraordinarily high Q values of 108-108 lead to high energy density, narrow resonant-wavelength lines and a lengthy cavity ringdown. They can also be coated in order to better control their properties or to increase their functionality. Their very high sensitivity to changes in the surrounding medium has been exploited for several sensing applications: protein adsorption, trace gas detection, impurity detection in liquids, structural health monitoring of composite materials, detection of electric fields, pressure sensing, and so on. In the present paper, after a general introduction to WGM resonators, attention is focused on spherical microresonators, either in bulk or in bubble format, to their fabrication, characterization and functionalization. The state of the art in the area of biosensing is presented, and the perspectives of further developments are discussed.

Optical Microbubble Resonators with High Refractive Index Inner Coating for Bio-Sensing Applications: An Analytical Approach.

The design of Whispering Gallery Mode Resonators (WGMRs) used as an optical transducer for biosensing represents the first and crucial step towards the optimization of the final device performance in terms of sensitivity and Limit of Detection (LoD). Here, we propose an analytical method for the design of an optical microbubble resonator (OMBR)-based biosensor. In order to enhance the OMBR sensing performance, we consider a polymeric layer of high refractive index as an inner coating for the OMBR. The effect of this layer and other optical/geometrical parameters on the mode field distribution, sensitivity and LoD of the OMBR is assessed and discussed, both for transverse electric (TE) and transverse magnetic (TM) polarization. The obtained results do provide physical insights for the development of OMBR-based biosensor.

Resonance Frequency of Optical Microbubble Resonators: Direct Measurements and Mitigation of Fluctuations.

This work shows the improvements in the sensing capabilities and precision of an Optical Microbubble Resonator due to the introduction of an encaging poly(methyl methacrylate) (PMMA) box. A frequency fluctuation parameter σ was defined as a score of resonance stability and was evaluated in the presence and absence of the encaging system and in the case of air- or water-filling of the cavity. Furthermore, the noise interference introduced by the peristaltic and the syringe pumping system was studied. The measurements showed a reduction of σ in the presence of the encaging PMMA box and when the syringe pump was used as flowing system.

Generation of hyper-parametric oscillations in silica microbubbles.

Cavity resonant enhanced stimulated Raman scattering (SRS), four-wave mixing, and broadband hyper-parametric oscillation in silica microbubble whispering gallery mode resonators (WGMR) in forward and backward directions are reported in this Letter. We show that microbubbles can operate not only in a highly ideal two-photon emission regime, but also generate combs, both natively and multi-mode spaced. The nonlinear process is phase matched because of the interaction of different mode families of the resonator.

Stimulated anti-Stokes Raman scattering resonantly enhanced in silica microspheres.

Efficient stimulated anti-Stokes Raman scattering has been observed in silica microspherical resonators pumped by CW laser. This process is observed in the normal dispersion regime. The lack of correlation between stimulated anti-Stokes and Stokes scattering spectra indicates that the signal has to be resonant with the cavity.

Toxic effects of amyloid fibrils on cell membranes: the importance of ganglioside GM1.

The interaction of amyloid aggregates with the cell plasma membrane is currently considered among the basic mechanisms of neuronal dysfunction in amyloid neurodegeneration. We used amyloid oligomers and fibrils grown from the yeast prion Sup35p, responsible for the specific prion trait [PSI(+)], to investigate how membrane lipids modulate fibril interaction with the membranes of cultured H-END cells and cytotoxicity. Sup35p shares no homology with endogenous mammalian polypeptide chains. Thus, the generic toxicity of amyloids and the molecular events underlying cell degeneration can be investigated without interference with analogous polypeptides encoded by the cell genome. Sup35 fibrils bound to the cell membrane without increasing its permeability to Ca(2+). Fibril binding resulted in structural reorganization and aggregation of membrane rafts, with GM1 clustering and alteration of its mobility. Sup35 fibril binding was affected by GM1 or its sialic acid moiety, but not by cholesterol membrane content, with complete inhibition after treatment with fumonisin B1 or neuraminidase. Finally, cell impairment resulted from caspase-8 activation after Fas receptor translocation on fibril binding to the plasma membrane. Our observations suggest that amyloid fibrils induce abnormal accumulation and overstabilization of raft domains in the cell membrane and provide a reasonable, although not unique, mechanistic and molecular explanation for fibril toxicity.

Ancient mitochondrial genome diversity in South America: Contributions from Quebrada del Toro, Northwestern Argentina.

OBJECTIVES: The objective of this study was to enhance our understanding of the population history in South America, specifically Northwestern Argentina, by analyzing complete ancient mitogenomes of individuals from the Ojo de Agua archeological site (970 BP) in Quebrada del Toro (Salta, Argentina). MATERIALS AND METHODS: We analyzed teeth from four individuals from the site Ojo de Agua (970 ± 60 BP), located in Quebrada del Toro (Andean region of Northwestern Argentina). DNA extracts were converted to double-stranded DNA libraries and indexed using unique dual-indexing primer combinations. DNA libraries were then enriched for the complete mitochondrial genome, pooled at equimolar concentrations, and sequenced on an Illumina® MiSeq™ platform. Reads from high quality libraries were trimmed, merged, and then mapped to the revised Cambridge Reference Sequence. The aDNA damage patterns were assessed and contamination estimated. Finally, variants were called, filtered, and the consensus mitogenome was constructed and used for haplogroup assignment. We also compiled available mitogenome sequences from ancient and present-day populations from the Southcentral Andes and other surrounding regions in Argentina. Maximum Likelihood and Bayesian phylogenetic reconstructions were obtained using the generated dataset. RESULTS: We successfully obtained the complete mitogenome sequence from one individual with an average depth coverage of 102X. We discovered a novel haplotype that was assigned to haplogroup D1. Phylogenetic reconstructions suggests that this haplotype falls within the sister branches of the D1j lineage, forming a well-supported clade. The estimate TMRCA of this clade that includes D1j and its sister branches ranged between 12,535 and 18,669 ya. DISCUSSION: The sequence analyzed in this study represents the first ancient mitogenome from within the valley region in Northwestern Argentina. We found that a representative of a lineage highly associated with D1j was already present approximately 1000 BP in the region. Our results agree with the proposed origin of D1j in other regions north of Patagonia and independent of the Pacific coast fast migratory route, contrary to what was originally hypothesized. This study highlights the lack of information regarding pre-Hispanic genetic diversity and contributes to the knowledge about the peopling process in South America.

OBJETIVOS: El objetivo de este estudio fue contribuir a mejorar la comprensión del poblamiento de Sudamérica, específicamente del Noroeste Argentino, mediante el análisis de mitogenomas antiguos completos de individuos del sitio arqueológico Ojo de Agua (970 AP), Quebrada del Toro (Salta, Argentina). MATERIALES Y MÉTODOS: Se analizaron dientes de cuatro individuos del sitio Ojo de Agua (970 ± 60 AP), ubicado en la Quebrada del Toro (región andina del Noroeste Argentino). A partir de los extractos de ADN se armaron librerías doble cadena y se indexaron utilizando combinaciones únicas de pares de cebadores. Las librerías fueron luego enriquecidas en ADN mitocondrial, llevadas a concentraciones equimolares y secuenciadas en una plataforma Illumina® MiSeq™. Las lecturas de las librerías de alta calidad fueron recortadas, fusionadas y, posteriormente, alineadas con la Secuencia de Referencia de Cambridge revisada. Se evaluaron los patrones de daño del ADNa y se estimó la contaminación. Por último, se identificaron las variantes, se filtraron y se construyó el mitogenoma consenso, que se utilizó para la asignación de haplogrupos. Además, se recopilaron secuencias de mitogenomas disponibles para poblaciones pre-hispánicas y actuales de los Andes Centrosur y otras regiones adyacentes de Argentina. Utilizando el conjunto de datos generado, se obtuvieron reconstrucciones filogenéticas mediante Máxima Verosimilitud y estimación Bayesiana. RESULTADOS: Se obtuvo la secuencia completa del mitogenoma de un individuo con una profundidad media de 102X. Esta secuencia corresponde a un nuevo haplotipo que fue asignado al haplogrupo D1. Las reconstrucciones filogenéticas sugirieron que este haplotipo se encuentra dentro del grupo hermano del linaje D1j, conformando un clado bien soportado. La datación estimada para este clado que contiene a D1j y a todo su grupo hermano fue de entre 12.535 y 18.669 años atrás. DISCUSIÓN: La secuencia analizada en este estudio representa el primer mitogenoma antiguo de la región valliserrana del Noroeste Argentino. Se halló que un representante de un linaje altamente asociado con D1j ya estaba presente hace aproximadamente 1000 años AP en la región. Nuestros resultados concuerdan con un origen de D1j en otras regiones al norte de la Patagonia, e independientemente de la ruta migratoria rápida por la costa del Pacífico, en contraposición a lo planteado inicialmente. Este estudio pone en evidencia la falta de información que se tiene actualmente sobre la diversidad genética en tiempos prehispánicos y contribuye al conocimiento del proceso de poblamiento de Sudamérica.

Performance of Eudragit coated whispering gallery mode resonator-based immunosensors.

Whispering gallery mode resonators (WGMR) are an efficient tool for the realization of optical biosensors. A high Q factor preservation is a crucial requirement for good biosensor performances. In this work we present an Eudragit®L100 coated microspherical WGMR as an efficient immunosensor. The developed resonator was morphologically characterized using fluorescence microscopy. The functionalization process was tuned to preserve the high Q factor of the resonator. The protein binding assay was optically characterized in terms of specificity in buffer solution.

Hybrid microspheres for nonlinear Kerr switching devices.

Electronic Kerr effect in a polyfluorene derivative is used to reversibly switch near infrared probe beam resonantly coupled to a hybrid polymer-silica microspherical resonator. NIR pumping at 780 nm in pulsed laser regime is used for non-linear switching of the WGM resonances that shift as much as 2 GHz for 50 mW of average pump power, compared to a shift of 250 MHz for the same average pump power at CW regime. The absence of temporal drift and the magnitude of this shift confirm the Kerr nature of the switching, ruling out thermooptical effects.

Optical microspherical resonators for biomedical sensing.

Optical resonators play an ubiquitous role in modern optics. A particular class of optical resonators is constituted by spherical dielectric structures, where optical rays are total internal reflected. Due to minimal reflection losses and to potentially very low material absorption, these guided modes, known as whispering gallery modes, can confer the resonator an exceptionally high quality factor Q, leading to high energy density, narrow resonant-wavelength lines and a lengthy cavity ringdown. These attractive characteristics make these miniaturized optical resonators especially suited as laser cavities and resonant filters, but also as very sensitive sensors. First, a brief analysis is presented of the characteristics of microspherical resonators, of their fabrication methods, and of the light coupling techniques. Then, we attempt to overview some of the recent advances in the development of microspherical biosensors, underlining a number of important applications in the biomedical field.

Supercontinuum source tuned by an on-axis monochromator for fluorescence lifetime imaging.

We report on the application of an optically tunable coherent white light source, based on the supercontinuum generation from microstructured optical fibres, to fluorescence lifetime imaging (FLIM) microscopy and Foerster resonance energy transfer (FRET). A prototype lens based on monotonic longitudinal chromatic aberration is used for tuning the supercontinuum wavelengths from 400 to 1000 nm and acts as an axial monochromator, suitable for fibre delivery in confocal microscopy.

Parametrical Optomechanical Oscillations in PhoXonic Whispering Gallery Mode Resonators.

We report on the experimental and theoretical analysis of parametrical optomechanical oscillations in hollow spherical phoxonic whispering gallery mode resonators due to radiation pressure. The optically excited acoustic eigenmodes of the phoxonic cavity oscillate regeneratively leading to parametric oscillation instabilities.

Nonlinear immunofluorescent assay for androgenic hormones based on resonant structures.

We report for the first time the use of two photon fluorescence as detection method of affinity binding reactions. We use a resonant grating waveguide structure as platform enhancement for detecting the interaction between fluorescent labeled Boldenone, a non-natural androgenic hormone, and a specific anti-anabolic antibody. We were able to detect a surface coverage of approximately 0.7 ng/mm(2).

Efficient frequency generation in phoXonic cavities based on hollow whispering gallery mode resonators.

We report on nonlinear optical effects on phoxonic cavities based on hollow whispering gallery mode resonators pumped with a continuous wave laser. We observed stimulated scattering effects such as Brillouin and Raman, Kerr effects such as degenerated and non-degenerated four wave mixing, and dispersive wave generation. These effects happened concomitantly. Hollow resonators give rise to a very rich nonlinear scenario due to the coexistence of several family modes.

Aptasensors Based on Whispering Gallery Mode Resonators.

In this paper, we review the literature on optical evanescent field sensing in resonant cavities where aptamers are used as biochemical receptors. The combined advantages of highly sensitive whispering gallery mode resonator (WGMR)-based transducers, and of the unique properties of aptamers make this approach extremely interesting in the medical field, where there is a particularly high need for devices able to provide real time diagnosis for cancer, infectious diseases, or strokes. However, despite the superior performances of aptamers compared to antibodies and WGMR to other evanescent sensors, there is not much literature combining both types of receptors and transducers. Up to now, the WGMR that have been used are silica microspheres and silicon oxynitride (SiON) ring resonators.

Stimulated Stokes and Antistokes Raman Scattering in Microspherical Whispering Gallery Mode Resonators.

Dielectric microspheres can confine light and sound for a length of time through high quality factor whispering gallery modes (WGM). Glass microspheres can be thought as a store of energy with a huge variety of applications: compact laser sources, highly sensitive biochemical sensors and nonlinear phenomena. A protocol for the fabrication of both the microspheres and coupling system is given. The couplers described here are tapered fibers. Efficient generation of nonlinear phenomena related to third order optical non-linear susceptibility Χ((3)) interactions in triply resonant silica microspheres is presented in this paper. The interactions here reported are: Stimulated Raman Scattering (SRS), and four wave mixing processes comprising Stimulated Anti-stokes Raman Scattering (SARS). A proof of the cavity-enhanced phenomenon is given by the lack of correlation among the pump, signal and idler: a resonant mode has to exist in order to obtain the pair of signal and idler. In the case of hyperparametric oscillations (four wave mixing and stimulated anti-stokes Raman scattering), the modes must fulfill the energy and momentum conservation and, last but not least, have a good spatial overlap.