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.

Photostability of Contrast Agents for Photoacoustics: The Case of Gold Nanorods.

Plasmonic particles as gold nanorods have emerged as powerful contrast agents for critical applications as the photoacoustic imaging and photothermal ablation of cancer. However, their unique efficiency of photothermal conversion may turn into a practical disadvantage, and expose them to the risk of overheating and irreversible photodamage. Here, we outline the main ideas behind the technology of photoacoustic imaging and the use of relevant contrast agents, with a main focus on gold nanorods. We delve into the processes of premelting and reshaping of gold nanorods under illumination with optical pulses of a typical duration in the order of few ns, and we present different approaches to mitigate this issue. We undertake a retrospective classification of such approaches according to their underlying, often implicit, principles as: constraining the initial shape; or speeding up their thermal coupling to the environment by lowering their interfacial thermal resistance; or redistributing the input energy among more particles. We discuss advantages, disadvantages and contexts of practical interest where one solution may be more appropriate than the other.

A bionic shuttle carrying multi-modular particles and holding tumor-tropic features.

The systemic delivery of composite nanoparticles remains an outstanding challenge in cancer nanomedicine, and the principal reason is a complex interplay of biological barriers. In this regard, adaptive cell transfer may represent an alternative solution to circumvent these barriers down to the tumor microenvironment. Here, tumor-tropic macrophages are proposed as a tool to draw and vehiculate modular nanoparticles integrating magnetic and plasmonic components. The end result is a bionic shuttle that exhibits a plasmonic band within the so-called therapeutic window arising from as much as 40 pg Au per cell, magnetization in the order of 150 pemu per cell, and more than 90% of the pristine viability and chemotactic activity of its biological component, until at least two days of preparation. Its synergistic combination of plasmonic, magnetic and tumor-tropic functions is assessed in vitro for applications as magnetic guidance or sorting, with a propulsion around 4 µm s-1 for a magnetic gradient of 0.8 T m-1, the optical hyperthermia of cancer, with stability of photothermal conversion to temperatures exceeding 50∘C, and the photoacoustic imaging of cancer under realistic conditions. These results collectively suggest that a bionic design may be a promising roadmap to reconcile the efforts for multifunctionality and targeted delivery, which are both key goals in nanomedicine.

Optically activated and interrogated plasmonic hydrogels for applications in wound healing.

We disclose the use of hybrid materials featuring Au/Ag core/shell nanorods in porous chitosan/polyvinyl alcohol scaffolds for applications in tissue engineering and wound healing. The combination of Au and Ag in a single construct provides synergistic opportunities for optical activation of functions as near infrared laser tissue bonding, and remote interrogation to return parameters of prognostic relevance in wound healing monitoring. In particular, the bimetallic component ensures optical tunability, enhanced shelf life and photothermal stability, serves as a reservoir of germicidal silver cations, and changes in near-infrared and visible color according to the environmental level of oxidative stress. At the same time, the polymeric blend is ideal to bind connective tissue upon photothermal activation, and to support fabrication processes that provide high porosity, such as electrospinning, thus putting all the premises for cellular repopulation and antimicrobial protection.

Impact of Kapitza resistance on the stability and efficiency of photoacoustic conversion from gold nanorods.

Plasmonic particles have been proposed for a broad variety of optical and hybrid applications, including the photothermal ablation and photoacoustic imaging of cancer, or their integration in photonic sensors. Here, we address the effect of thermal resistance at the gold-water interface, or Kapitza resistance, on the performance of photoacoustic conversion of gold nanorods. Our findings point to possible strategies for the optimization of plasmonic particles as contrast agents for imaging, or even as transducers for biosensing. We perform numerical simulations that project a simultaneous increase of efficiency and stability of photoacoustic conversion with a decrease of Kapitza resistance. We suggest an effective approach to modulate Kapitza resistance by including underresolved features as roughness or the presence of adsorbates. Inspired by this idea, we synthesize a rough variant of gold nanorods by the deposition and galvanic replacement of a silver shell, where roughness provides higher photoacoustic signals by about 70% and damage thresholds by 120%. In addition, we coat our particles with a protein corona and find a decrease of photoacoustic signals with shell thickness, which may inspire new solutions for biosensors based on a mechanism of photoacoustic transduction. Both our findings are consistent with an effective modulation of Kapitza resistance, which decreases upon roughening, due to an underlying increase of specific surface area, and increases upon coating with a protein shell that may act as a thermal insulation. We discuss possible directions to gain more advantage of our concept for topical applications at the crossroads of plasmonics, biomedical optics and biosensing.

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.

Fluorescence-Sensor Mapping for the in Vineyard Non-Destructive Assessment of Crimson Seedless Table Grape Quality.

Non-destructive tools for the in situ evaluation of vine fruit quality and vineyard management can improve the market value of table grape. We proposed a new approach based on a portable fluorescence sensor to map the ripening level of Crimson Seedless table grape in five different plots in the East, Central-North and South of the Macedonia Region of Greece. The sensor provided indices of ripening and color such as SFRR and ANTHRG correlated to the chlorophyll and anthocyanin berry contents, respectively. The mean ANTHRG index was significantly different among all the plots examined due to the occurrence of different environmental conditions and/or asynchronous ripening processes. The indices presented moderate, poor in some cases, spatial variability, probably due to a significant vine-to-vine, intra-vine and intra-bunch variability. The cluster analysis was applied to the plot with the most evident spatial structure (at Kilkis). Krigged maps of the SFRR, ANTHRG and yield were classified by k-means clustering in two-zones that differed significantly in their mean values. ANTHRG and SFRR were inversely correlated over 64% of the plot. SFRR appeared to be a potential useful proxy of yield since it was directly correlated to yield over 66% of the plot. The grape color (ANTHRG) was slightly higher over the low-yield zones with respect to the high-yield zones. Our study showed that the combination of anthocyanins and chlorophyll indices detected in the field on Crimson Seedless table grape by a portable fluorescence sensor can help in defining the best harvest time and the best areas for harvesting.

Hybrid organosilicon/polyol phantom for photoacoustic imaging.

The rapid development of hardware and software for photoacoustic technologies is urging the establishment of dedicated tools for standardization and performance assessment. In particular, the fabrication of anatomical phantoms for photoacoustic imaging remains an open question, as current solutions have not yet gained unanimous support. Here, we propose that a hybrid material made of a water-in-oil emulsion of glycerol and polydimethylsiloxane may represent a versatile platform to host a broad taxonomy of hydrophobic and hydrophilic dyes and recapitulate the optical and acoustic features of bio tissue. For a full optical parameterization, we refer to Wróbel, et al. [ Biomed. Opt. Express7, 2088 (2016)], where this material was first presented for optical imaging. Instead, here, we complete the picture and find that its speed of sound and acoustic attenuation resemble those of pure polydimethylsiloxane, i.e. respectively 1150 ± 30 m/s and 3.5 ± 0.4 dB/(MHz·cm). We demonstrate its use under a commercial B-mode scanner and a home-made A-mode stage for photoacoustic analysis to retrieve the ground-truth encoded in a multilayer architecture containing indocyanine green, plasmonic particles and red blood cells. Finally, we verify the stability of its acoustic, optical and geometric features over a time span of three months.

1064-nm-resonant gold nanorods for photoacoustic theranostics within permissible exposure limits.

Therapeutic and diagnostic methods based on photomechanical effects are attracting much current attention in contexts as oncology, cardiology and vascular surgery, for such applications as photoacoustic imaging or microsurgery. Their underlying mechanism is the generation of ultrasound or cavitation from the interaction of short optical pulses with endogenous dyes or targeted contrast agents. Among the latter, gold nanorods are outstanding candidates, but their use has mainly been reported for photoacoustic imaging and photothermal treatments. Conversely, much less is still known about their value as a precision tool for photomechanical manipulations, such as to impart local damage with high spatial resolution through the expansion and collapse of microbubbles. Here, we address the feasibility of gold nanorods exhibiting a distribution of surface plasmon resonances between about 900 to above 1100 nm as a contrast agent for photoacoustic theranostics. After testing their cytotoxicity and cellular uptake, we discuss their photostability and use to mediate cavitation and the photomechanical destruction of targeted cells. We find that the choice of a plasmonic band peaking around 1064 nm is key to enhance the translational potential of this approach. With respect to the standard alternative of 800 nm, at 1064 nm, relevant regulations on optical exposure are less restrictive and the photonic technology is more mature.

A multifunctional organosilica cross-linker for the bio-conjugation of gold nanorods.

We report on the use of organosilica shells to couple gold nanorods to functional peptides and modulate their physiochemical and biological profiles. In particular, we focus on the case of cell penetrating peptides, which are used to load tumor-tropic macrophages and implement an innovative drug delivery system for photothermal and photoacoustic applications. The presence of organosilica exerts subtle effects on multiple parameters of the particles, including their size, shape, electrokinetic potential, photostability, kinetics of endocytic uptake and cytotoxicity, which are investigated by the interplay of colorimetric methods and digital holographic microscopy. As a rule of thumb, as the thickness of organosilica increases from none to ∼30nm, we find an improvement of the photophysical performances at the expense of a deterioration of the biological parameters. Therefore, detailed engineering of the particles for a certain application will require a careful trade-off between photophysical and biological specifications.

Preparation and Photoacoustic Analysis of Cellular Vehicles Containing Gold Nanorods.

Gold nanorods are attractive for a range of biomedical applications, such as the photothermal ablation and the photoacoustic imaging of cancer, thanks to their intense optical absorbance in the near-infrared window, low cytotoxicity and potential to home into tumors. However, their delivery to tumors still remains an issue. An innovative approach consists of the exploitation of the tropism of tumor-associated macrophages that may be loaded with gold nanorods in vitro. Here, we describe the preparation and the photoacoustic inspection of cellular vehicles containing gold nanorods. PEGylated gold nanorods are modified with quaternary ammonium compounds, in order to achieve a cationic profile. On contact with murine macrophages in ordinary Petri dishes, these particles are found to undergo massive uptake into endocytic vesicles. Then these cells are embedded in biopolymeric hydrogels, which are used to verify that the stability of photoacoustic conversion of the particles is retained in their inclusion into cellular vehicles. We are confident that these results may provide new inspiration for the development of novel strategies to deliver plasmonic particles to tumors.

Organosilicon phantom for photoacoustic imaging.

Photoacoustic imaging is an emerging technique. Although commercially available photoacoustic imaging systems currently exist, the technology is still in its infancy. Therefore, the design of stable phantoms is essential to achieve semiquantitative evaluation of the performance of a photoacoustic system and can help optimize the properties of contrast agents. We designed and developed a polydimethylsiloxane (PDMS) phantom with exceptionally fine geometry; the phantom was tested using photoacoustic experiments loaded with the standard indocyanine green dye and compared to an agar phantom pattern through polyethylene glycol-gold nanorods. The linearity of the photoacoustic signal with the nanoparticle number was assessed. The signal-tonoiseratio and contrast were employed as image quality parameters, and enhancements of up to 50 and up to 300%, respectively, were measured with the PDMS phantom with respect to the agar one. A tissue-mimicking (TM)-PDMS was prepared by adding TiO2 and India ink; photoacoustic tests were performed in order to compare the signal generated by the TM-PDMS and the biological tissue. The PDMS phantom can become a particularly promising tool in the field of photoacoustics for the evaluation of the performance of a PA system and as a model of the structure of vascularized soft tissues.

Graphene as a photothermal switch for controlled drug release.

Graphene has recently emerged as a novel material in the biomedical field owing to its optical properties, biocompatibility, large specific surface area and low cost. In this paper, we provide the first demonstration of the possibility of using light to remotely trigger the release of drugs from graphene in a highly controlled manner. Different drugs including chemotherapeutics and proteins are firmly adsorbed onto reduced graphene oxide (rGO) nanosheets dispersed in a biopolymer film and then released by individual millisecond-long light pulses generated by a near infrared (NIR) laser. Here graphene plays the dual role of a versatile substrate for temporary storage of drugs and an effective transducer of NIR-light into heat. Drug release appears to be narrowly confined within the size of the laser spot under noninvasive conditions and can be precisely dosed depending on the number of pulses. The approach proposed paves the way for tailor-made pharmacological treatments of chronic diseases, including cancer, anaemia and diabetes.

Hybrid nanocomposite films for laser-activated tissue bonding.

We report new advancements in the biomedical exploitation of plasmonic nanoparticles as an effective platform for the photothermal repair of biological tissue. Chitosan films are loaded with gold nanorods with intense optical absorption in the "therapeutic window" of deepest light penetration through the body, and then activated by near infrared laser excitation to give adhesion with adjacent connective tissues. The adhesion consists of 0.07 mm(2) welds of ~20 kPa tensile strength at the film/tissue interface, which are obtained by administration of pulses with duration in the hundreds of millisecond timescale from a diode laser at ~130 J cm(-2). We investigate the adhesive effect as a function of pulse power and duration and identify an optimal operative window to achieve effective and reproducible welds with minimal detrimental superheating. These results may prove valuable to standardize laser bonding techniques and meet current needs for new knowledge which is urged by the penetration of nanotechnology into biomedical optics.

Micro-photoluminescence of GaAs/AlGaAs triple concentric quantum rings.

A systematic optical study, including micro, ensemble and time resolved photoluminescence of GaAs/AlGaAs triple concentric quantum rings, self-assembled via droplet epitaxy, is presented. Clear emission from localized states belonging to the ring structures is reported. The triple rings show a fast decay dynamics, around 40 ps, which is expected to be useful for ultrafast optical switching applications.