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Herein, we demonstrate a cavity-enhanced hyperspectral refractometric imaging using an all-dielectric photonic crystal slab (PhCS). Our approach takes advantage of the synergy between two mechanisms, surface-enhanced fluorescence (SEF) and refractometric sensing, both based on high-Q resonances in proximity of bound states in the continuum (BICs). The enhanced local optical field of the first resonance amplifies of 2 orders of magnitude the SEF emission of a probe dye. Simultaneously, hyperspectral refractometric sensing, based on Fano interference between second mode and fluorescence emission, is used for mapping the spatially variant refractive index produced by the specimen on the PhCS. The spectral matching between first resonance and input laser is modulated by the specimen local refractive index, and thanks to the calibrated dependence with the spectral shift of the Fano resonance, the cavity tuning is used to achieve an enhanced correlative refractometric map with a resolution of 10-5 RIU within femtoliter-scale sampling volumes. This is experimentally applied also on live prostate cancer cells grown on the PhCS, reconstructing enhanced surface refractive index images at the single-cell level. This dual mechanism of quasi-BIC spatially variant gain tracked by quasi-BIC refractometric sensing provides a correlative imaging platform that can find application in many fields for monitoring physical and biochemical processes, such as molecular interactions, chemical reactions, or surface cell analysis.

Several living organisms are able to synthesize complex nanostructures provided with peculiar physical and chemical properties by means of finely-tuned, genetically controlled biomineralization processes. Frustules, in particular, are micro- and nano-structured silica shells produced by ubiquitous diatom microalgae, whose optical properties have been recently exploited in photonics, solar energy harvesting, and biosensing. Metallization of diatom biosilica, both in the shape of intact frustules or diatomite particles, can trigger plasmonic effects that in turn can find application in high-sensitive detection platforms, allowing to obtain effective nanosensors at low cost and on a large scale. The aim of the present review article is to provide a wide, complete overview on the main metallization techniques applied to diatom biosilica and on the principal applications of diatom-based plasmonic devices mainly but not exclusively in the fields of biochemical sensing, diagnostics and therapeutics.

Profilin 1 –encoded by PFN1– is a small actin-binding protein with a tumour suppressive role in various adenocarcinomas and pagetic osteosarcomas. However, its contribution to tumour development is not fully understood. Using fix and live cell imaging, we report that Profilin 1 inactivation results in multiple mitotic defects, manifested prominently by anaphase bridges, multipolar spindles, misaligned and lagging chromosomes, and cytokinesis failures. Accordingly, next-generation sequencing technologies highlighted that Profilin 1 knock-out cells display extensive copy-number alterations, which are associated with complex genome rearrangements and chromothripsis events in primary pagetic osteosarcomas with Profilin 1 inactivation. Mechanistically, we show that Profilin 1 is recruited to the spindle midzone at anaphase, and its deficiency reduces the supply of actin filaments to the cleavage furrow during cytokinesis. The mitotic defects are also observed in mouse embryonic fibroblasts and mesenchymal cells deriving from a newly generated knock-in mouse model harbouring a Pfn1 loss-of-function mutation. Furthermore, nuclear atypia is also detected in histological sections of mutant femurs. Thus, our results indicate that Profilin 1 has a role in regulating cell division, and its inactivation triggers mitotic defects, one of the major mechanisms through which tumour cells acquire chromosomal instability.

Optical tweezers are tools made of light that enable contactless pushing, trapping, and manipulation of objects ranging from atoms to space light sails. Since the pioneering work by Arthur Ashkin in the 1970s, optical tweezers have evolved into sophisticated instruments and have been employed in a broad range of applications in life sciences, physics, and engineering. These include accurate force and torque measurement at the femtonewton level, microrheology of complex fluids, single micro- and nanoparticle spectroscopy, single-cell analysis, and statistical-physics experiments. This roadmap provides insights into current investigations involving optical forces and optical tweezers from their theoretical foundations to designs and setups. It also offers perspectives for applications to a wide range of research fields, from biophysics to space exploration., Comment: 181 pages, 61 figures

Surface-enhanced Raman spectroscopy (SERS) has become a powerful tool for biosensing applications owing to its fingerprint recognition, high sensitivity, multiplex detection, and biocompatibility. This review provides an overview of the most significant aspects of SERS for biomedical and biosensing applications. We first introduced the mechanisms at the basis of the SERS amplifications: electromagnetic and chemical enhancement. We then illustrated several types of substrates and fabrication methods, with a focus on gold-based nanostructures. We further analyzed the relevant factors for the characterization of the SERS sensor performances, including sensitivity, reproducibility, stability, sensor configuration (direct or indirect), and nanotoxicity. Finally, a representative selection of applications in the biomedical field is provided.

In the last few years, gold nanoparticles (AuNPs) have been widely used in biomedical applications; unfortunately, in some cases, they induce toxic effects on biological systems. Among these, the activation of the innate immune system (inflammatory response) is considered a central issue for assessing health risks. Although the origin of nanotoxicity is not well known, the cause could be associated with the presence of contaminants on nanoparticles surface, such as bacterial endotoxin. The bacterial endotoxin, also known as Lipopolysaccharide (LPS), is the main component of gram-negative bacteria cell walls and is considered one of the major contaminants in the environment. Therefore, AuNPs can be easily contaminated during the synthesis process or their manipulation, and nanotoxicological assessment has to be performed before using AuNPs for biomedical purposes. Traditional assays for LPS detection are not always effective because AuNPs could interfere with other components and alter the final readout. Surface-enhanced Raman spectroscopy (SERS) is a powerful tool for the analysis of molecules on metallic NPs, allowing to detect low concentration LPS with excellent sensitivity and reproducibility. In this work, we demonstrate that SERS technique can detect low amount of LPS (fg) on AuNPs (50 nm); moreover, we studied the internalization of bare and LPS-coated AuNPs in human monocyte-derived macrophages, by Raman Imaging, and their inflammatory effect by in vitro evaluation of pro and anti-inflammatory cytokine production.

Here, we present a multimodal imaging approach based on Raman spectroscopy and PSDH imaging to detect circulating tumor cells. Particularly, the ability of tumor cells to internalize glucose faster than normal cells was investigated. Deuterated-glucose was used as Raman vibrational tag since its signal is present in the “silent” zone of Raman cell spectra. Raman information were correlated with the visualization of the cell birefringence by PSDH imaging. Each technique provides different information which combination gives a molecular and morphological signature of each cell, allowing to discriminate between normal and cancer cells with a sensitivity and specificity up to 80%.

A novel hyperspectral sensing imaging taking advantage of engineered all-dielectric metasurfaces supporting bound states in the continuum here is discussed. This approach combines surface-enhanced fluorescence and res- onant shift both based on high-Q resonances in proximity of bound states in the continuum. The amplification of the optical field on resonance allows increasing the fluorescence emission of a dye as a function of the spatial- variant dielectric environment in the near-field of the structure. We first demonstrate the fluorescence emission amplification by resonant pump matching in microscopy configuration. Then, we take advantage of Fano reso- nances in the fluorescence emission to map the spatially variant environment of biological cells. To demonstrate the real implementation of the proposed BIC-enhanced imaging as a platform for biosensing, hyperspectral maps of prostate cancer cells are experimentally reconstructed.

Surface-enhanced Raman scattering (SERS) has established itself as powerful tool for molecular sensing in biology and medicine. The integration of SERS systems with optical fiber is a challenging but potentially very rewarding endeavour. However, efforts to transfer the technology from the laboratory to the clinic have been frustrated by the lack of robust stable and sensitive substrates on the fiber tip, as well as the complexity of interfacing between sample and the substrate itself. Here, we propose the Lab-on-Fiber SERS optrodes, realized on the optical fiber tip by nanosphere lithography. Three types of highly ordered and reproducible SERS-active substrates have been realized: close-packed array (CPA); CPA after sphere removal (SR) and sparse array (SA) of polystyrene nanospheres, covered by a gold thin layer. To optimize the SERS probes, we compared the SERS performances in terms of Enhancement Factor (EF) and reproducibility pertaining to different patterns with different nanosphere diameters and gold thicknesses using the biphenyl-4-thiol (BPT), as target molecule. Moreover, we analysed and compared the SERS spectra of two representative biological probes, bovine serum albumin (BSA, medium molecule) and red blood cells (RBCs), in order to correlate the SERS response to the morphology and hysteric hindrance of the biological target. The SERS analysis indicated that the CPA substrate amplifies the BPT Raman intensity twice as well as the SR and SA substrates, while BSA and RBCs, with the CPA substrate, provide signals comparable to those of SR and SA substrates. Finally, we have optimized a Raman system for SERS optrode operation with efficient lighting and collection via optical fiber.

The interaction between engineered nanoparticles and the bacterial lipopolysaccharide, or endotoxin, is an event that warrants attention. Endotoxin is one of the most potent stimulators of inflammation and immune reactions in human beings, and is a very common contaminant in research labs. In nanotoxicology and nanomedicine, the presence of endotoxin on the nanoparticle surface affects their biological properties leading to misinterpretation of results. This review discusses the importance of detecting the endotoxin contamination on nanoparticles, focusing on the current method of endotoxin detection and their suitability for nanoparticulate materials. Conversely, the capacity of nanoparticles to bind endotoxin can be enhanced by functionalization with endotoxin-capturing molecules, opening the way to the development of novel endotoxin detection assays., The authors were supported by the EU H2020 grants ENDONANO [GA 812661] and PANDORA [GA 671881], the IG grant [21420] of the Associazione Italiana Ricerca sul Cancro (AIRC), the project NeON [ARS01_00769] of the Program PRIN, and the projects CIRO- Campania Infrastructure for Research in Oncology and PLATT of the Programma Operativo Regionale of Regione Campania [POR FERS 2014/2020].

Nanoscale delivery systems have been investigated for therapy due to their advantages, including the sustained delivery of drugs to cells and reduction of systemic toxicity compared to conventional treatments. However, their application is still hampered by experimental challenges, such as the investigation of the drug release in cells rather than in vitro. Here, we describe a hybrid nanoplatform for monitoring the drug release in living colorectal cancer (CRC) cells by Surface-Enhanced Raman Scattering (SERS). Specifically, the anticancer drug Galunisertib is encapsulated in diatomite nanoparticles (DNPs) decorated by gold nanoparticles (AuNPs) and capped by gelatin. The combination of DNP loading capacities with the Raman enhancement of Galunisertib provided by AuNPs enables bio-imaging and drug delivery without using fluorophores or markers, avoiding fluorescence-quenching issues. Thanks to the Raman enhancement of Galunisertib, the drug release profile is monitored and quantified in living cells by SERS with a femtogram scale resolution. When the gelatin shell is digested by proteases, Galunisertib is released and its SERS spectrum decreases, allowing real-time quantification in CRC cells. The therapeutic efficiency of the Galunisertib delivery platform offers an alternative route for lowering drug dose and toxicity.

The small molecule Galunisertib (LY2157299, LY) shows multiple anticancer activities blocking the transforming growth factor-β1 receptor, responsible for the epithelial-to-mesenchymal transition (EMT) by which colorectal cancer (CRC) cells acquire migratory and metastatic capacities. However, frequent dosing of LY can produce highly toxic metabolites. Alternative strategies to reduce drug side effects can rely on nanoscale drug delivery systems that have led to a medical revolution in the treatment of cancer, improving drug efficacy and lowering drug toxicity. Here, a hybrid nanosystem (DNP-AuNPs-LY@Gel) made of a porous diatomite nanoparticle decorated with plasmonic gold nanoparticles, in which LY is retained by a gelatin shell, is proposed. The multifunctional capability of the nanosystem is demonstrated by investigating the efficient LY delivery, the enhanced EMT reversion in CRCs and the intracellular quantification of drug release with a sub-femtogram resolution by surface-enhanced Raman spectroscopy (SERS). The LY release trigger is the pH sensitivity of the gelatin shell to the CRC acidic microenvironment. The drug release is real-time monitored at single-cell level by analyzing the SERS signals of LY in CRC cells. The higher efficiency of LY delivered by the DNP-AuNPs-LY@Gel complex paves the way to an alternative strategy for lowering drug dosing and consequent side effects.

Understanding the modes of interaction between human monocytes/macrophages and engineered nanoparticles is the basis for assessing particle safety, in terms of activation of innate/inflammatory reactions, and their possible exploitation for medical applications. In vitro assessment of nanoparticle-macrophage interaction allows for examining the response of primary human cells, but the conventional 2D cultures do not reproduce the three-dimensional spacing of a tissue and the interaction of macrophages with the extracellular tissue matrix, conditions that shape macrophage recognition capacity and reactivity. Here, we have compared traditional 2D cultures with cultures on a 3D collagen matrix for evaluating the capacity gold nanoparticles to induce monocyte activation and subsequent innate memory in human blood monocytes in comparison to bacterial LPS. Results show that monocytes react to stimuli almost in the same way in 2D and 3D cultures in terms of production of TNFα and IL-6, but that notable differences are found when IL-8 and IL-1Ra are examined, in particular in the recall/memory response of primed cells to a second stimulation, with the 3D cultures showing cell activation and memory effects of nanoparticles better. In addition, the response variations in monocytes/macrophages from different donors point towards a personalized assessment of the nanoparticle effects on macrophage activation., This research was funded by the European Commission FP7 project HUMUNITY (GA 316383), the Horizon 2020 projects PANDORA (GA 671881) and ENDONANO (GA 812661), the Italian MIUR Flagship InterOmics project MEMORAT and the PRIN project 20173ZECCM.

In the last decade, Raman Spectroscopy (RS) was demonstrated to be a label-free, non-invasive and non-destructive optical spectroscopy allowing the improvement in diagnostic accuracy in cancer and analytical assessment for cell sensing. This review discusses how Raman spectra can lead to a deeper molecular understanding of the biochemical changes in cancer cells in comparison to non-cancer cells, analyzing two key examples, leukemia and breast cancer. The reported Raman results provide information on cancer progression and allow the identification, classification, and follow-up after chemotherapy treatments of the cancer cells from the liquid biopsy. The key obstacles for RS applications in cancer cell diagnosis, including quality, objectivity, number of cells and velocity of the analysis, are considered. The use of multivariant analysis, such as principal component analysis (PCA) and linear discriminate analysis (LDA), for an automatic and objective assessment without any specialized knowledge of spectroscopy is presented. Raman imaging for cancer cell mapping is shown and its advantages for routine clinical pathology practice and live cell imaging, compared to single-point spectral analysis, are debated. Additionally, the combination of RS with microfluidic devices and high-throughput screening for improving the velocity and the number of cells analyzed are also discussed. Finally, the combination of the Raman microscopy (RM) with other imaging modalities, for complete visualization and characterization of the cells, is described.

© 2020 by the authors., Innate immune memory is characterized by a modulation in the magnitude with which innate immune cells such as monocytes and macrophages respond to potential dangers, subsequent to previous exposure to the same or unrelated agents. In this study, we have examined the capacity of gold nanoparticles (AuNP), which are already in use for therapeutic and diagnostic purposes, to modulate the innate memory induced by bacterial agents. The induction of innate memory was achieved in vitro by exposing human primary monocytes to bacterial agents (lipopolysaccharide -LPS-, or live Bacille Calmette-Guérin -BCG) in the absence or presence of AuNP. After the primary activation, cells were allowed to return to a resting condition, and eventually re-challenged with LPS. The induction of memory was assessed by comparing the response to the LPS challenge of unprimed cells with that of cells primed with bacterial agents and AuNP. The response to LPS was measured as the production of inflammatory (TNFα, IL-6) and anti-inflammatory cytokines (IL-10, IL-1Ra). While ineffective in directly inducing innate memory per se, and unable to influence LPS-induced tolerance memory, AuNP significantly affected the memory response of BCG-primed cells, by inhibiting the secondary response in terms of both inflammatory and anti-inflammatory factor production. The reprogramming of BCG-induced memory towards a tolerance type of reactivity may open promising perspectives for the use of AuNP in immunomodulatory approaches to autoimmune and chronic inflammatory diseases., This research was funded by the EU Commission H2020 projects PANDORA (GA 671881) and ENDONANO (GA 812661), the Italian MIUR Flagship InterOmics project MEMORAT, and the Cluster project Medintech (CNT01_00177_962865).

A new, simple digital holography-based polarization microscope for quantitative birefringence imaging of biological cells is presented. As a proof of concept, two different class of cells have been characterized by polarization sensitive digital holographic imaging (PSDHI). These two cases study reported are: differentiation of leukaemia cells and identification of reacted sperm cells. Although further experimentation is necessary, the suggested approach could represent a prospective label-free diagnostic tool for use in biological and medical research and diagnosis.

Leukemia diagnosis and classification requires the use and combination of several technologies usually involving staining and morphology examining of cells in a blood sample, or selective detection of specific cell membrane antigens. Standard morphological protocols often lack the specificity and sensitivity required for an accurate and early diagnosis. Fluorescence based analyses of the cell surface receptors, although providing specific information, are limited by photobleaching and number of surface proteins that can be imaged simultaneously. In this review, we consider our recent efforts towards the development of a Raman spectroscopy-based protocol for a complete biochemical and morphological analysis of leukemia cells. Indeed, Raman spectroscopy is an optical technique based on inelastic scattering of light by vibrating molecules and can provide high specific biochemical information with minimal or no sample pretreatment. We discuss single-point Raman analysis for the identification, classification and follow-up of acute lymphoblastic leukemia type B (B-ALL). Normal B cells and three B-ALL cell lines (RS4;11, REH and MN60) were analyzed, and several intrinsic Raman markers associated with proteins, nucleic acids and lipids were used for cells sensing and follow-up after the low-dose chemotherapy treatment. Multivariate statistical analyses allowed us confirming the significance of the Raman data for identifying and classifying the leukemia cells in their maturation/differentiation stages. False color Raman imaging was additionally used to visualize the leukemia cell morphology, as the standard May Grunwald-Giemsa histochemical staining protocol, but without the use of any label. Finally, the selective probing of leukemia cell membrane by surface enhanced Raman scattering (SERS) is reported. The SERS substrate was a 3D diatom frustule covered with a gold layer (40 nm in thickness). These observations are promising for the development of a Raman-based protocol for the diagnosis and the complete morpho-functional assessment of leukemias.

Plasmonic substrates play a crucial role in the confinement and manipulation of localized electromagnetic fields at the nanoscale. The large electromagnetic field enhancement at metal/dielectric interfaces is widely exploited in surface-enhanced fluorescence (SEF) and surface-enhanced Raman scattering (SERS) spectroscopies. Despite the advantage of near-field enhancement, unfortunately, in metals, the large absorption at optical frequencies induces local heating of the analyte fluid with possible damage of the biological material. In addition, in SEF plasmonic substrates, spacer layers are necessary to minimize undesired fluorescence quenching due to nonradiative decay, which strongly depends on the distance between molecules and metallic substrates. Therefore, the possibility of managing surface electromagnetic states mimicking surface-plasmon resonances in terms of spatial localization, high-field intensity, and dispersion characteristics, while avoiding metallic losses is of great interest. However, dielectric nanoantennas can currently provide limited possibilities in the visible range of optical frequencies. We present the realization of all-dielectric metasurfaces made of nanostructured transparent silicon nitride supporting bound states in the continuum (BICs). We show that this special kind of Fano resonances can be effectively used in standard microscopy for practical applications. We achieved concurrent enhancements of ~103 fold of fluorescence emission and Raman scattering far-field intensities of molecules dispersed on these metasurfaces. In addition, we demonstrate that the gain of conventional SERS signals can be increased by more than one order of magnitude by resonant matching of the localized surface plasmon resonance with the BIC field. Our results can find significant applications for enhanced sensing, Raman imaging, and nonlinear processes.

In this work, we propose the use of complex, bioderived nanostructures as efficient surface-enhanced Raman scattering (SERS) substrates for chemical analysis of cellular membranes. These structures were directly obtained from a suitable gold metalization of the Pseudonitzchia multistriata diatom silica shell (the so called frustule), whose grating-like geometry provides large light coupling with external radiation, whereas its extruded, subwavelength lateral edge provides an excellent interaction with cells without steric hindrance. We carried out numerical simulations and experimental characterizations of the supported plasmonic resonances and optical near-field amplification. We thoroughly evaluated the SERS substrate enhancement factor as a function of the metalization parameters and finally applied the nanostrucures for discriminating cell membrane Raman signals. In particular, we considered two cases where the membrane composition plays a fundamental role in the assessment of several pathologies, that is, red blood cells and B-leukemia REH cells.

In this paper, we studied the influence of the geometrical configuration of surface-enhanced Raman scattering (SERS)-active self-assembling gold nanostructures on the interaction with biological targets of different sizes, aiming at large-scope SERS lab-on-fiber optrodes composed of highly efficient SERS substrates integrated onto optical fibre tips. By using nanosphere lithography, we fabricated three types of highly ordered and reproducible SERS-active substrates. To correlate the SERS response to the steric hindrance of the biological target, we experimentally analysed and compared the SERS spectra of three representative biological probes, i.e., ultralow-molecular-weight molecules of biphenyl-4-thiol (BPT, small molecule, 186.27 Da), bovine serum albumin (BSA, medium molecule, 66.5 kDa) and red blood cells (RBCs, diameter

This work presents the use of a Raman-based device and its integration/correlation with ultrasensitive microscopies (surface-enhanced Raman microscopy and confocal microscopy) for the molecular imaging and classification of normal lymphocytic B-cell and different leukemia cells from peripheral blood samples. The label-free Raman approach allowed for categorization of leukocytes into their respective lineage based on the biochemical information provided by Raman spectra. Furthermore, the single-point Raman analysis permitted the accurate discrimination of normal B-cells and leukemia-transformed cell lines (RS4;11, REH, MN60 cells), and more significantly, between leukemia cells in different differentiation/maturation stages. Raman imaging successfully and without cell destruction provided non-destructively information on the cell morphology, similarly to the standard May Grunwald-Giemsa histochemical staining protocol, but without the requirements for sample preparation. Finally, the specific analysis of the leukemic cell membrane by surface enhanced Raman scattering is shown. The reported results are promising for the development of a Raman-based protocol for the diagnosis and the complete morphofunctional assessment of cancer cells.

The localization of the electromagnetic field at the nanoscale can play a key role in many applications, such as sensing, spectroscopy and energy conversion. In the last years, great efforts have been performed to study and realize all-dielectric loss-free nanostructures to confine the radiation without the limits imposed by plasmonic systems. It has been demonstrated that an all-dielectric photonic crystal (PhC) metasurface can support bound states in the continuum (BICs) - resonant states of infinite lifetime - due to the interaction between trapped electromagnetic modes. Experimentally, this involves very narrow coupled resonances, with a high Q-factor and a possible extremely large field intensity enhancement, up to 6 orders of magnitude. Here, we demonstrate that the field enhancement in proximity of the surface can be explored to boost lightmatter interaction in spectroscopic sensing. We design and realize an innovative sensing scheme for bulk and surface measurement with ultra-high figure of merit for the recognition of protein-protein interaction and the detection of low molecular weight molecules. In addition, we design a dielectric dual scheme based on metasurfaces supporting BICs to amplify fluorescence emission and Raman scattering of probe molecules dispersed on the surface of the PhC. Our results provide new solutions for light manipulation at the nanoscale, especially for sensing and nonlinear optics applications.

Raman microspectroscopy (RM) and polarization sensitive digital holographic imaging (PSDHI) are valuable analytical tools in biological and medical research, allowing the detection of both biochemical and morphological variations of the sample without labels or long sample preparation. Here, using this multi-modal approach we analyze in vitro human sperm capacitation and the acrosome reaction induced by heparin. The multimodal microscopy provides morphofunctional information that can assess the sperms ability to respond to capacitation stimuli (sperm function). More precisely, the birefringence analysis in sperm cells can be used as an indicator of its structural normality. Indeed, digital holography applied for polarization imaging allows for revelation of the polarization state of the sample, showing a total birefringence of the sperm head in non-reacted spermatozoa, and a birefringence localized in the post-acrosomal region in reacted spermatozoa. Additionally, RM allows the detection and spectroscopic characterization of protein/lipid delocalization in the plasma and acrosomal membranes that can be used as valuable Raman biomarkers of sperm function. Interestingly, these spectral variations can be correlated with different time phases of the cell capacitation response. Although further experimentation is required, the proposed multimodal approach could represent a potential label-free diagnostic tool for use in reproductive medicine and the diagnosis of infertility.

We propose a new, simple digital holography-based polarization microscope for birefringence imaging of biological cells. Although further experimentation is required, the proposed approach could represent a potential label-free diagnostic tool for use in biological and medical research and diagnosis.

We report on our activities related to the development of surface enhanced Raman scattering (SERS) probes realized onto the optical fiber tip (OFT) through nanosphere lithography. In the first stage of our research, we adapted the nanosphere lithography to operate on the optical fiber tip, by assessing the process and demonstrating either the potentiality or the repeatability of the proposed nanopatterning approach. Successively, we investigated the ability of the manufactured structures on the fiber tip to act as SERS probes by measuring the SERS spectra in presence of a Biphenyl Thiol (BPT) monolayer. Firstly, we focused the attention on the samples shaped as closed packed array of nanospheres covered by gold. The analysis allowed us to identify the most promising SERS platform, exhibiting an Enhancement Factor (EF) of 4x10(5) and a SERS measurements variability lower than 10%. We addressed also the limitations related to the use of the same optical fiber for both illumination and light collection by selecting a commercial optical fiber exhibiting a suitable trade-off in terms of high excitation/collection efficiency and low silica background. Current activities are devoted to the investigation of other nanopatterns on the optical fiber tip (namely, Sparse Array of metallo-dielectric spheres) and the analysis of the probes response against different molecules.

Moving the polarization of the incident wave along a meridian of the Poincare sphere, experimentally we show that the coupling with the fundamental Bloch’s surface waves of the mode, provide a spatially coherent, macroscopic spinmomentum locked propagation along the symmetry axes of the PhCM. This novel mechanism of light-spin manipulation enables a versatile implementation of spin-optical structures that may pave the way to novel strategies for light spin technology and photonic multiplatform implementations.

Coupling digital holography with high-specific Raman spectroscopy is an attractive means of identifying biochemical changes in cells by both physical topography and spectroscopy. We have used this approach to simultaneously study biochemical and morphological characteristics of human sperm cells irradiated with green laser radiation. Severe spermatozoa variations associated with a topological redistribution of the sample and a gradual decrease in the Raman signal intensity were detected in a label-free configuration. Importantly, at laser fluences where no morphological alterations were detected (30 MJ/cm2 ), high specific spectral variations were monitored to evaluate the cell photodegradation.

The localization of the electromagnetic field at the nanoscale can play a key role in many applications, such as sensing, spectroscopy and energy conversion. In the last years, great efforts have been performed to study and realize all-dielectric loss-free nanostructures to confine the radiation without the limits imposed by the plasmonic systems. Here we demonstrate that the field enhancement in proximity of a photonic crystal metasurface supporting bound states in the continuum can be explored to boost the light-matter interaction. We design and realize an innovative sensing scheme for bulk and surface measurement with ultra-high figure of merit and apply this new configuration for studying a specific protein-protein interaction. The recognition scheme can be coupled to a fluorescence-based sensing approach, which exploits the capability of the sensor to strongly enhance fluorescence signals. Our results provide new solutions for light manipulation at the nanoscale, especially for sensing and nonlinear optics applications.

Diatoms can represent the major component of phytoplankton and contribute massively to global primary production in the oceans. Over tens of millions of years they developed an intricate porous silica shell, the frustule, which ensures mechanical protection, sorting of nutrients from harmful agents, and optimization of light harvesting. Several groups of microalgae evolved different strategies of protection towards ultraviolet radiation (UVR), which is harmful for all living organisms mainly through the formation of dimeric photoproducts between adjacent pyrimidines in DNA. Even in presence of low concentrations of UV-absorbing compounds, several diatoms exhibit significant UVR tolerance. We here investigated the mechanisms involved in UVR screening by diatom silica investments focusing on single frustules of a planktonic centric diatom, Coscinodiscus wailesii, analyzing absorption by the silica matrix, diffraction by frustule ultrastructure and also UV conversion into photosynthetically active radiation exerted by nanostructured silica photoluminescence. We identified the defects and organic residuals incorporated in frustule silica matrix which mainly contribute to absorption; simulated and measured the spatial distribution of UVR transmitted by a single valve, finding that it is confined far away from the diatom valve itself; furthermore, we showed how UV-to-blue radiation conversion (which is particularly significant for photosynthetic productivity) is more efficient than other emission transitions in the visible spectral range. Diatoms can represent the major component of phytoplankton and contribute massively to global primary production in the oceans. Over tens of millions of years they developed an intricate porous silica shell, the frustule, which ensures mechanical protection, sorting of nutrients from harmful agents, and optimization of light harvesting. Several groups of microalgae evolved different strategies of protection towards ultraviolet radiation (UVR), which is harmful for all living organisms mainly through the formation of dimeric photoproducts between adjacent pyrimidines in DNA. Even in presence of low concentrations of UV-absorbing compounds, several diatoms exhibit significant UVR tolerance. We here investigated the mechanisms involved in UVR screening by diatom silica investments focusing on single frustules of a planktonic centric diatom, Coscinodiscus wailesii, analyzing absorption by the silica matrix, diffraction by frustule ultrastructure and also UV conversion into photosynthetically active radiation exerted by nanostructured silica photoluminescence. We identified the defects and organic residuals incorporated in frustule silica matrix which mainly contribute to absorption; simulated and measured the spatial distribution of UVR transmitted by a single valve, finding that it is confined far away from the diatom valve itself; furthermore, we showed how UV-to-blue radiation conversion (which is particularly significant for photosynthetic productivity) is more efficient than other emission transitions in the visible spectral range.