Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines.

BACKGROUND: Breast cancer (BC) is a heterogeneous neoplasm characterized by several subtypes. One of the most aggressive with high metastasis rates presents overexpression of the human epidermal growth factor receptor 2 (HER2). A quantitative evaluation of HER2 levels is essential for a correct diagnosis, selection of the most appropriate therapeutic strategy and monitoring the response to therapy. RESULTS: In this paper, we propose the synergistic use of SERS and Raman technologies for the identification of HER2 expressing cells and its accurate assessment. To this end, we selected SKBR3 and MDA-MB-468 breast cancer cell lines, which have the highest and lowest HER2 expression, respectively, and MCF10A, a non-tumorigenic cell line from normal breast epithelium for comparison. The combined approach provides a quantitative estimate of HER2 expression and visualization of its distribution on the membrane at single cell level, clearly identifying cancer cells. Moreover, it provides a more comprehensive picture of the investigated cells disclosing a metabolic signature represented by an elevated content of proteins and aromatic amino acids. We further support these data by silencing the HER2 gene in SKBR3 cells, using the RNA interference technology, generating stable clones further analysed with the same combined methodology. Significant changes in HER2 expression are detected at single cell level before and after HER2 silencing and the HER2 status correlates with variations of fatty acids and downstream signalling molecule contents in the context of the general metabolic rewiring occurring in cancer cells. Specifically, HER2 silencing does reduce the growth ability but not the lipid metabolism that, instead, increases, suggesting that higher fatty acids biosynthesis and metabolism can occur independently of the proliferating potential tied to HER2 overexpression. CONCLUSIONS: Our results clearly demonstrate the efficacy of the combined SERS and Raman approach to definitely pose a correct diagnosis, further supported by the data obtained by the HER2 gene silencing. Furthermore, they pave the way to a new approach to monitor the efficacy of pharmacologic treatments with the aim to tailor personalized therapies and optimize patients' outcome.

An Innovative Fiber-Optic Hydrophone for Seismology: Testing Detection Capacity for Very Low-Energy Earthquakes.

An innovative fiber-optic hydrophone (FOH) was developed and investigated via an experiment at sea; it is capable of operating at a very low frequency of the seismic spectrum and detecting small magnitude earthquakes. The FOH exploits an optical fiber coil wrapped around a sensitive mandrel in a Michelson interferometric configuration. The FOH operated for about seven days at a water depth of 40 m, in the Campi Flegrei volcanic area (Southern Italy), and a few meters from a well-calibrated PZT hydrophone used as a reference. Thirty-three local earthquakes occurred during the simultaneous operation of the two hydrophones, allowing a straightforward comparison of the recordings. The local earthquakes occurred at an epicentral distance less than 2.5 km from the site of recording, and were estimated to be in the range of magnitude from -0.8 to 2.7. The analysis of the recorded earthquake waveforms in the frequency and time domains allowed retrieving the response function of the FOH in the frequency range from 5 to 70 Hz. The FOH responsivity in terms of acoustic pressure reached about 230 nm/Pa and was flat in the studied frequency range. Due to the high quality of the FOH recordings, this equipment is suitable for applications addressing submarine volcanic activity and the background seismicity of active faults in the ocean.

Innovative Photonic Sensors for Safety and Security, Part III: Environment, Agriculture and Soil Monitoring.

In order to complete this set of three companion papers, in this last, we focus our attention on environmental monitoring by taking advantage of photonic technologies. After reporting on some configurations useful for high precision agriculture, we explore the problems connected with soil water content measurement and landslide early warning. Then, we concentrate on a new generation of seismic sensors useful in both terrestrial and under water contests. Finally, we discuss a number of optical fiber sensors for use in radiation environments.

Innovative Photonic Sensors for Safety and Security, Part II: Aerospace and Submarine Applications.

The employability of photonics technology in the modern era's highly demanding and sophisticated domain of aerospace and submarines has been an appealing challenge for the scientific communities. In this paper, we review our main results achieved so far on the use of optical fiber sensors for safety and security in innovative aerospace and submarine applications. In particular, recent results of in-field applications of optical fiber sensors in aircraft monitoring, from a weight and balance analysis to vehicle Structural Health Monitoring (SHM) and Landing Gear (LG) monitoring, are presented and discussed. Moreover, underwater fiber-optic hydrophones are presented from the design to marine application.

Innovative Photonic Sensors for Safety and Security, Part I: Fundamentals, Infrastructural and Ground Transportations.

Our group, involving researchers from different universities in Campania, Italy, has been working for the last twenty years in the field of photonic sensors for safety and security in healthcare, industrial and environment applications. This is the first in a series of three companion papers. In this paper, we introduce the main concepts of the technologies employed for the realization of our photonic sensors. Then, we review our main results concerning the innovative applications for infrastructural and transportation monitoring.

Lab-On-Fiber Technology: A Roadmap toward Multifunctional Plug and Play Platforms.

This review presents an overview of the "lab-on-fiber technology" vision and the main milestones set in the technological roadmap to achieve the ultimate objective of developing flexible, multifunctional plug and play fiber-optic platforms designed for specific applications. The main achievements, obtained with nanofabrication strategies for unconventional substrates, such as optical fibers, are discussed here. The perspectives and challenges that lie ahead are highlighted with a special focus on full spatial control at the nanoscale and high-throughput production scenarios. The rapid progress in the fabrication stage has opened new avenues toward the development of multifunctional plug and play platforms, discussed here with particular emphasis on new functionalities and unparalleled figures of merit, to demonstrate the potential of this powerful technology in many strategic application scenarios. The paper also analyses the benefits obtained from merging lab-on-fiber (LOF) technology objectives with the emerging field of optomechanics, especially at the microscale and the nanoscale. We illustrate the main advances at the fabrication level, describe the main achievements in terms of functionalities and performance, and highlight future directions and related milestones. All achievements reviewed and discussed clearly suggest that LOF technology is much more than a simple vision and could play a central role not only in scenarios related to diagnostics and monitoring but also in the Information and Communication Technology (ICT) field, where optical fibers have already yielded remarkable results.

Nanosphere Lithography on Fiber: Towards Engineered Lab-On-Fiber SERS Optrodes.

In this paper we report on the engineering of repeatable surface enhanced Raman scattering (SERS) optical fiber sensor devices (optrodes), as realized through nanosphere lithography. The Lab-on-Fiber SERS optrode consists of polystyrene nanospheres in a close-packed arrays configuration covered by a thin film of gold on the optical fiber tip. The SERS surfaces were fabricated by using a nanosphere lithography approach that is already demonstrated as able to produce highly repeatable patterns on the fiber tip. In order to engineer and optimize the SERS probes, we first evaluated and compared the SERS performances in terms of Enhancement Factor (EF) pertaining to different patterns with different nanosphere diameters and gold thicknesses. To this aim, the EF of SERS surfaces with a pitch of 500, 750 and 1000 nm, and gold films of 20, 30 and 40 nm have been retrieved, adopting the SERS signal of a monolayer of biphenyl-4-thiol (BPT) as a reliable benchmark. The analysis allowed us to identify of the most promising SERS platform: for the samples with nanospheres diameter of 500 nm and gold thickness of 30 nm, we measured values of EF of 4 × 105, which is comparable with state-of-the-art SERS EF achievable with highly performing colloidal gold nanoparticles. The reproducibility of the SERS enhancement was thoroughly evaluated. In particular, the SERS intensity revealed intra-sample (i.e., between different spatial regions of a selected substrate) and inter-sample (i.e., between regions of different substrates) repeatability, with a relative standard deviation lower than 9 and 15%, respectively. Finally, in order to determine the most suitable optical fiber probe, in terms of excitation/collection efficiency and Raman background, we selected several commercially available optical fibers and tested them with a BPT solution used as benchmark. A fiber probe with a pure silica core of 200 µm diameter and high numerical aperture (i.e., 0.5) was found to be the most promising fiber platform, providing the best trade-off between high excitation/collection efficiency and low background. This work, thus, poses the basis for realizing reproducible and engineered Lab-on-Fiber SERS optrodes for in-situ trace detection directed toward highly advanced in vivo sensing.

Lab-on-fiber technology: a new vision for chemical and biological sensing.

The integration of microfluidics and photonic biosensors has allowed achievement of several laboratory functions in a single chip, leading to the development of photonic lab-on-a-chip technology. Although a lot of progress has been made to implement such sensors in small and easy-to-use systems, many applications such as point-of-care diagnostics and in vivo biosensing still require a sensor probe able to perform measurements at precise locations that are often hard to reach. The intrinsic property of optical fibers to conduct light to a remote location makes them an ideal platform to meet this demand. The motivation to combine the good performance of photonic biosensors on chips with the unique advantages of optical fibers has thus led to the development of the so-called lab-on-fiber technology. This emerging technology envisages the integration of functionalized materials on micro- and nano-scales (i.e. the labs) with optical fibers to realize miniaturized and advanced all-in-fiber probes, especially useful for (but not limited to) label-free chemical and biological applications. This review presents a broad overview of lab-on-fiber biosensors, with particular reference to lab-on-tip platforms, where the labs are integrated on the optical fiber facet. Light-matter interaction on the fiber tip is achieved through the integration of thin layers of nanoparticles or nanostructures supporting resonant modes, both plasmonic and photonic, highly sensitive to local modifications of the surrounding environment. According to the physical principle that is exploited, different configurations - such as localized plasmon resonance probes, surface enhanced Raman scattering probes and photonic probes - are classified, while various applications are presented in context throughout. For each device, the surface chemistry and the related functionalization protocols are reviewed. Moreover, the implementation strategies and fabrication processes, either based on bottom-up or top-down approaches, are discussed. In conclusion we highlight some of the further development opportunities, including lab-in-a-needle technology, which could have a direct and disruptive impact in localized cancer treatment applications.

An intrusion detection system for the protection of railway assets using Fiber Bragg Grating sensors.

We demonstrate the ability of Fiber Bragg Gratings (FBGs) sensors to protect large areas from unauthorized activities in railway scenarios such as stations or tunnels. We report on the technological strategy adopted to protect a specific depot, representative of a common scenario for security applications in the railway environment. One of the concerns in the protection of a railway area centers on the presence of rail-tracks, which cannot be obstructed with physical barriers. We propose an integrated optical fiber system composed of FBG strain sensors that can detect human intrusion for protection of the perimeter combined with FBG accelerometer sensors for protection of rail-track access. Several trials were carried out in indoor and outdoor environments. The results demonstrate that FBG strain sensors bonded under a ribbed rubber mat enable the detection of intruder break-in via the pressure induced on the mat, whereas the FBG accelerometers installed under the rails enable the detection of intruders walking close to the railroad tracks via the acoustic surface waves generated by footsteps. Based on a single enabling technology, this integrated system represents a valuable intrusion detection system for railway security and could be integrated with other sensing functionalities in the railway field using fiber optic technology.

Assessment of Primary Human Liver Cancer Cells by Artificial Intelligence-Assisted Raman Spectroscopy.

We investigated the possibility of using Raman spectroscopy assisted by artificial intelligence methods to identify liver cancer cells and distinguish them from their Non-Tumor counterpart. To this aim, primary liver cells (40 Tumor and 40 Non-Tumor cells) obtained from resected hepatocellular carcinoma (HCC) tumor tissue and the adjacent non-tumor area (negative control) were analyzed by Raman micro-spectroscopy. Preliminarily, the cells were analyzed morphologically and spectrally. Then, three machine learning approaches, including multivariate models and neural networks, were simultaneously investigated and successfully used to analyze the cells' Raman data. The results clearly demonstrate the effectiveness of artificial intelligence (AI)-assisted Raman spectroscopy for Tumor cell classification and prediction with an accuracy of nearly 90% of correct predictions on a single spectrum.

Opto-acoustic behavior of coated fiber Bragg gratings.

In this paper, we present the study of the acousto-optic behavior of underwater-acoustic sensors constituted by fiber Bragg gratings (FBGs) coated by ring-shaped overlays. Via full-wave numerical simulations, we study the complex opto-acousto-mechanical interaction among an incident acoustic wave traveling in water, the optical fiber surrounded by the ring shaped coating, and the FBG inscribed the fiber, focusing on the frequency range 0.5-30 kHz of interest for SONAR applications. Our results fully characterize the mechanical behavior of an acoustically driven coated FBG, and highlight the key role played by the coating in enhancing significantly its sensitivity by comparison with a standard uncoated configuration. Furthermore, the hydrophone sensitivity spectrum exhibits characteristic resonances, which strongly improve the sensitivity with respect to its background (i.e., away from resonances) level. Via a three-dimensional modal analysis, we verify that the composite cylindrical structure of the sensor acts as an acoustic resonator tuned at the frequencies of its longitudinal vibration modes. In order to evaluate the sensor performance, we also carry out a comprehensive parametric analysis by varying the geometrical and mechanical properties of the coating, whose results also provide a useful design tool for performance optimization and/or tailoring for specific SONAR applications. Finally, a preliminary validation of the proposed numerical analysis has been carried out through experimental data obtained using polymeric coated FBGs sensors revealing a good agreement and prediction capability.

Tuning efficiency and sensitivity of guided resonances in photonic crystals and quasi-crystals: a comparative study.

In this paper, we present a comparative study of the tuning efficiency and sensitivity of guided resonances (GRs) in photonic crystal (PC) holed slabs based on periodic and aperiodically-ordered unit cells, aimed at assessing the applicability of these important technology platforms to ultra-compact optical sensors and active devices. In particular, with specific reference to square-lattice periodic PCs and aperiodically-ordered Ammann-Beenker photonic quasi-crystals, we study the effects of the hole radius, slab thickness, and refractive index on the GR sensitivity and tunability with respect to variation in the hole refractive index. Finally, we carry out a theoretical and numerical analysis in order to correlate the GR shift with the field distribution of the unperturbed (air holes) structures. Our results indicate that the spatial arrangement of the holes may strongly influence the tuning and sensitivity efficiency, and may provide new degrees of freedom and tools for the design and optimization of novel photonic devices for both sensing and telecommunication applications.

Experimental evidence of guided-resonances in photonic crystals with aperiodically ordered supercells.

We report on the first experimental evidence of guided resonances (GRs) in photonic crystal slabs based on aperiodically ordered supercells. Using Ammann-Beenker (quasiperiodic, eightfold symmetric) tiling geometry, we present our study on the fabrication, experimental characterization, and full-wave numerical simulation of two representative structures (with different filling parameters) operating at near-IR wavelengths (1300-1600 nm). Our results show a fairly good agreement between measurements and numerical predictions and pave the way for the development of new strategies (based on, e.g., the lattice symmetry breaking) for GR engineering.

Guided resonances in photonic crystals with point-defected aperiodically-ordered supercells.

In this paper, we study the excitation of guided resonances (GRs) in photonic-crystal slabs based on point-defected aperiodically-ordered supercells. With specific reference to perforated-slab structures and the Ammann-Beenker octagonal lattice geometry, we carry out full-wave numerical studies of the plane-wave responses and of the underlying modal structures, which illustrate the representative effects induced by the introduction of symmetry-preserving and symmetry-breaking defects. Our results demonstrate that breaking the supercell mirror symmetries via the judicious introduction of point-defects enables for the excitation of otherwise uncoupled GRs, with control on the symmetry properties of their field distributions, thereby constituting an attractive alternative to those GR-engineering approaches based on the asymmetrization of the hole shape. In this framework, aperiodically-ordered supercells seem to be inherently suited, in view of the variety of inequivalent defect sites that they can offer.

Fast and slow light in optical fibers through tilted fiber Bragg gratings.

In this paper, slow and fast light in optical fiber through tilted fiber Bragg grating (TFBG) are reported. The experimental results show the capability of TFBGs to enable group velocity control of an optical pulse in optical fiber, due to the anomalous dispersion features induced by the coupling between the propagating core mode and each counter-propagating coupling cladding mode. In particular, superluminal propagation of a pulse train has been observed at optical communication wavelengths with time advancements in the picoseconds time scale in 1cm long TFBG and group velocity as large as about two times the speed of light in optical fiber (approximately 1.3 x c0). Very good agreement has been obtained comparing the measured group delay of the TFBG with the one retrieved from the amplitude response through Hilbert transform. Finally, tunable slow and fast light has also been reported, demonstrating the possibility to control the group velocity at single wavelength through both fluidic and thermal actuation.

Guided resonances in photonic quasicrystals.

In this paper, we report on the first evidence of guided resonances (GRs) in aperiodically-ordered photonic crystals, tied to the concept of "quasicrystals" in solid-state physics. Via a full-wave numerical study of the transmittance response and the modal structure of a photonic quasicrystal (PQC) slab based on a representative aperiodic geometry (Ammann-Beenker octagonal tiling), we demonstrate the possibility of exciting GR modes, and highlight similarities and differences with the periodic case. In particular, we show that, as for the periodic case, GRs arise from the coupling of the incident plane-wave with degenerate modes of the PQC slab that exhibit a matching symmetry in the spatial distribution, and can still be parameterized via a Fano-like model. Besides the phenomenological implications, our results may provide new degrees of freedom in the engineering of GRs, and pave the way for new developments and applications.

Opto-mechanical lab-on-fibre seismic sensors detected the Norcia earthquake.

We have designed and developed lab-on-fibre seismic sensors containing a micro-opto-mechanical cavity on the fibre tip. The mechanical cavity is designed as a double cantilever suspended on the fibre end facet and connected to a proof mass to tune its response. Ground acceleration leads to displacement of the cavity length, which in turn can be remotely detected using an interferometric interrogation technique. After the sensors characterization, an experimental validation was conducted at the Italian National Institute of Geophysics and Volcanology (INGV), which is responsible for seismic surveillance over the Italian country. The fabricated sensors have been continuously used for long periods to demonstrate their effectiveness as seismic accelerometer sensors. During the tests, fibre optic seismic accelerometers clearly detected the seismic sequence that culminated in the severe Mw6.5 Norcia earthquake that struck central Italy on October 30, 2016. The seismic data provided by the optical sensors were analysed by specialists at the INGV. The wave traces were compared with state-of-the-art traditional sensors typically incorporated into the INGV seismic networks. The comparison verifies the high fidelity of the optical sensors in seismic wave detection, indicating their suitability for a novel class of seismic sensors to be employed in practical scenarios.

Nanosphere lithography for optical fiber tip nanoprobes.

This paper reports a simple and economical method for the fabrication of nanopatterned optical fiber nanotips. The proposed patterning approach relies on the use of the nanosphere lithography of the optical fiber end facet. Polystyrene (PS) nanospheres are initially self-assembled in a hexagonal array on the surface of water. The created pattern is then transferred onto an optical fiber tip (OFT). The PS monolayer colloidal crystal on the OFT is the basic building block that is used to obtain different periodic structures by applying further treatment to the fiber, such as metal coating, nanosphere size reduction and sphere removal. Ordered dielectric and metallo-dielectric sphere arrays, metallic nanoisland arrays and hole-patterned metallic films with feature sizes down to the submicron scale are achievable using this approach. Furthermore, the sizes and shapes of these periodic structures can be tailored by altering the fabrication conditions. The results indicate that the proposed self-assembly approach is a valuable route for the development of highly repeatable metallo-dielectric periodic patterns on OFTs with a high degree of order and low fabrication cost. The method can be easily extended to simultaneously produce multiple fibers, opening a new route to the development of fiber-optic nanoprobes. Finally, we demonstrate the effective application of the patterned OFTs as surface-enhanced Raman spectroscopy nanoprobes.