Interferometric Biosensor for High Sensitive Label-Free Recording of HiPS Cardiomyocytes Contraction in Vitro.

Heart disease remains a leading cause of global mortality, underscoring the need for advanced technologies to study cardiovascular diseases and develop effective treatments. We introduce an innovative interferometric biosensor for high-sensitivity and label-free recording of human induced pluripotent stem cell (hiPSC) cardiomyocyte contraction in vitro. Using an optical cavity, our device captures interference patterns caused by the contraction-induced displacement of a thin flexible membrane. First, we demonstrate the capability to quantify spontaneous contractions and discriminate between contraction and relaxation phases. We calculate a contraction-induced vertical membrane displacement close to 40 nm, which implies a traction stress of 34 ± 4 mN/mm2. Finally, we investigate the effects of a drug compound on contractility amplitude, revealing a significant reduction in contractile forces. The label-free and high-throughput nature of our biosensor may enhance drug screening processes and drug development for cardiac treatments. Our interferometric biosensor offers a novel approach for noninvasive and real-time assessment of cardiomyocyte contraction.

Ultrasound generation in water via quasi-periodically snapping polymeric core-shell micro-bead excited with radiowaves.

This work reports the results of a theoretical and numerical study showing the occurrence of stochastically resonating bistable dynamic in polymeric micro-bead of sub-micrometric size with stiff core and soft shell. The system, submerged in water, is excited with a pulsed laser working in the Mega-Hertz frequency range and tuned to match both an optical and acoustic resonance of the system. The laser interacts with the carbon nanotubes embedded in the shell of the polymeric micro-bead generating heat. The concurrent action of the generated heat with the standing acoustic oscillations, gives rise to a stochastically resonating bistable system. The system in fact is forced to switch between two states (identifiable with the creation and organized disruption of a quasi-hexagonal tessellation) via a snap-through-buckling mechanism. This phenomenon results in the unprecedented generation of pressure oscillations. These results open the way to develop a new type of core-shell micro-transducers for radioacoustic imaging applications able to work in the Mega-Hertz frequency range. From a more general thermodynamic perspective, the reported mechanism shows a remarkable periodicity and energy conversion efficiency.

Tsuchime-like Aluminum Film to Enhance Absorption in Ultra-Thin Photovoltaic Cells.

Ultra-thin solar cells enable materials to be saved, reduce deposition time, and promote carrier collection from materials with short diffusion lengths. However, light absorption efficiency in ultra-thin solar panels remains a limiting factor. Most methods to increase light absorption in ultra-thin solar cells are either technically challenging or costly, given the thinness of the functional layers involved. We propose a cost-efficient and lithography-free solution to enhance light absorption in ultra-thin solar cells-a Tsuchime-like self-forming nanocrater (T-NC) aluminum (Al) film. T-NC Al film can be produced by the electrochemical anodization of Al, followed by etching the nanoporous alumina. Theoretical studies show that T-NC film can increase the average absorbance by 80.3%, depending on the active layer's thickness. The wavelength range of increased absorption varies with the active layer thickness, with the peak of absolute absorbance increase moving from 620 nm to 950 nm as the active layer thickness increases from 500 nm to 10 µm. We have also shown that the absorbance increase is retained regardless of the active layer material. Therefore, T-NC Al film significantly boosts absorbance in ultra-thin solar cells without requiring expensive lithography, and regardless of the active layer material.

Plasmonic Bowl-Shaped Nanopore for Raman Detection of Single DNA Molecules in Flow-Through.

Plasmonic nanopores combined with Raman spectroscopy are emerging as platforms for single-molecule detection and sequencing in label-free mode. Recently, the ability of identifying single DNA bases or amino acids has been demonstrated for molecules adsorbed on plasmonic particles and then delivered into the plasmonic pores. Here, we report on bowl-shaped plasmonic gold nanopores capable of direct Raman detection of single λ-DNA molecules in a flow-through scheme. The bowl shape enables the incident laser to be focused into the nanopore to generate a single intense hot spot with no cut off in pore size. Therefore, we achieved ultrasmall focusing of NIR light in a spot of 3 nm. This enabled us to detect 7 consecutive bases along the DNA chain in flow-through conditions. Furthermore, we found a novel electrofluidic mechanism to manipulate the molecular trajectory within the pore volume so that the molecule is pushed toward the hot spot, thus improving the detection efficiency.

Passive Recording of Bioelectrical Signals from Non-Excitable Cells by Fluorescent Mirroring.

Bioelectrical variations trigger different cell responses, including migration, mitosis, and mutation. At the tissue level, these actions result in phenomena such as wound healing, proliferation, and pathogenesis. Monitoring these mechanisms dynamically is highly desirable in diagnostics and drug testing. However, existing technologies are invasive: either they require physical access to the intracellular compartments, or they imply direct contact with the cellular medium. Here, we present a novel approach for the passive recording of electrical signals from non-excitable cells adhering to 3D microelectrodes, based on optical mirroring. Preliminary results yielded a fluorescence intensity output increase of the 5,8% in the presence of a HEK-293 cell on the electrode compared to bare microelectrodes. At present, this technology may be employed to evaluate cell-substrate adhesion and monitor cell proliferation. Further refinements could allow extrapolating quantitative data on surface charges and resting potential to investigate the electrical phenomena involved in cell migration and cancer progression.

Binding of tyrosine kinase inhibitor to epidermal growth factor receptor: surface-enhanced infrared absorption microscopy reveals subtle protein secondary structure variations.

Surface-Enhanced Infrared Absorption (SEIRA) has been proposed as a valuable tool for protein binding studies, but its performances have been often proven on model proteins undergoing severe secondary structure rearrangements, while ligand binding only marginally involves the protein backbone in the vast majority of the biologically relevant cases. In this study we demonstrate the potential of SEIRA microscopy for highlighting the very subtle secondary structure modifications associated with the binding of Lapatinib, a tyrosine kinase inhibitor (TKI), to epidermal growth factor receptor (EGFR), a well-known driver of tumorigenesis in pathological settings such as lung, breast and brain cancers. By boosting the performances of Mid-IR plasmonic devices based on nanoantennas cross-geometry, accustoming the protein purification protocols, carefully tuning the protein anchoring methodology and optimizing the data analysis, we were able to detect EGFR secondary structure modification associated with few amino acids. A nano-patterned platform with this kind of sensitivity bridges biophysical and structural characterization methods, thus opening new possibilities in studying of proteins of biomedical interest, particularly for drug-screening purposes.

Mirroring Action Potentials: Label-Free, Accurate, and Noninvasive Electrophysiological Recordings of Human-Derived Cardiomyocytes.

The electrophysiological recording of action potentials in human cells is a long-sought objective due to its pivotal importance in many disciplines. Among the developed techniques, invasiveness remains a common issue, causing cytotoxicity or altering unpredictably cell physiological response. In this work, a new approach for recording intracellular signals of outstanding quality and with noninvasiveness is introduced. By taking profit of the concept of mirror charge in classical electrodynamics, the new proposed device transduces cell ionic currents into mirror charges in a microfluidic chamber, thus realizing a virtual mirror cell. By monitoring mirror charge dynamics, it is possible to effectively record the action potentials fired by the cells. Since there is no need for accessing or interacting with the cells, the method is intrinsically noninvasive. In addition, being based on optical recording, it shows high spatial resolution and high parallelization. As shown through a set of experiments, the presented methodology is an ideal candidate for the next generation devices for the reliable assessment of cardiotoxicity on human-derived cardiomyocytes. More generally, it paves the way toward a new family of in vitro biodevices that will lay a new milestone in the field of electrophysiology.

Breaking the symmetry of nanosphere lithography with anisotropic plasma etching induced by temperature gradients.

We report a novel anisotropic process, termed plasma etching induced by temperature gradients (PE-TG), which we use to modify the 3D morphology of a hexagonally close-packed polystyrene sphere array. Specifically, we combined an isotropic oxygen plasma (generated by a plasma cleaner) and a vertical temperature gradient applied from the bottom to the top of a colloidal mask to create an anisotropic etching process. As a result, an ordered array of well-defined and separated nano mushrooms is obtained. We demonstrate that the features of the mushrooms, namely the hat size and their intrinsic undercut, as well as the pillar diameter and height, can be easily tuned by adjusting the main parameters of the process i.e. the temperature gradient and etching time, or the spheres' size. We show that PS mushroom arrays can be used as nanostructured templates to fabricate plasmonic arrays, such as gold-capped nano mushrooms and ultra-small nanoapertures, by using vertical and oblique gold sputtering deposition respectively. PE-TG reveals a new, cheap and facile approach to produce plasmonic nanostructures of great interest in the fields of molecular sensing, surface-enhanced Raman scattering (SERS), energy harvesting and optoelectronics. We study the optical properties of the Au-capped nano mushroom arrays and their performance as biosensing platforms by performing SERS measurements.

Toward all on chip optical detection in the few molecule regime.

Integrated optics devices are one of the most promising technologies in many fields such as biosensing, optical monitoring, and portable devices. They provide several advantages such as unique sensitivity and the possibility of the well-established and developed silicon photonics technology. However some challenges still remain open, as the implementation of multiplex assay able to reach the single particle sensitivity. In this context, we propose a new design for a Si-based photonic structure that enables the realization of on chip sub-wavelength optical sources. The idea is based on the insertion of opportunely designed nanometric holes in the photonic circuit, which are available for analyte detection with high efficiency. We propose three different configurations in which both excitation and detection are obtained through the same waveguide thus simplifying the detection scheme and potentially enabling multiplexed detection. We proved the high confinement of the electromagnetic field in the holes both by theoretical modelling and spectroscopic measurements. We investigate the possibility of inserting an arbitrary number of optical sources by using a resonator and evaluate advantages and drawbacks of resonating and non-resonating solutions. Finally, we report the proof-of-concept experiment, where detection sensitivity down to single Quantum Dots is obtained by combining the novel design with fluorescence-based techniques. Importantly, the presented results are achieved by a simple modification of photonic sensing chips which are already on the market thus having an excellent translational perspective.

Microfluidic Multielectrode Arrays for Spatially Localized Drug Delivery and Electrical Recordings of Primary Neuronal Cultures.

Neuropathological models and neurological disease progression and treatments have always been of great interest in biomedical research because of their impact on society. The application of in vitro microfluidic devices to neuroscience-related disciplines provided several advancements in therapeutics or neuronal modeling thanks to the ability to control the cellular microenvironment at spatiotemporal level. Recently, the introduction of three-dimensional nanostructures has allowed high performance in both in vitro recording of electrogenic cells and drug delivery using minimally invasive devices. Independently, both delivery and recording have let to pioneering solutions in neurobiology. However, their combination on a single chip would provide further fundamental improvements in drug screening systems and would offer comprehensive insights into pathologies and diseases progression. Therefore, it is crucial to develop platforms able to monitor progressive changes in electrophysiological behavior in the electrogenic cellular network, induced by spatially localized injection of biochemical agents. In this work, we show the application of a microfluidic multielectrode array (MEA) platform to record spontaneous and chemically stimulated activity in primary neuronal networks. By means of spatially localized caffeine injection via microfluidic nanochannels, the device demonstrated its capability of combined localized drug delivery and cell signaling recording. The platform could detect activity of the neural network at multiple sites while delivering molecules into just a few selected cells, thereby examining the effect of biochemical agents on the desired portion of cell culture.

Multiplexed Discrimination of Single Amino Acid Residues in Polypeptides in a Single SERS Hot Spot.

The SERS-based detection of protein sequences with single-residue sensitivity suffers from signal dominance of aromatic amino acid residues and backbones, impeding detection of non-aromatic amino acid residues. Herein, we trap a gold nanoparticle in a plasmonic nanohole to generate a single SERS hot spot for single-molecule detection of 2 similar polypeptides (vasopressin and oxytocin) and 10 distinct amino acids that constitute the 2 polypeptides. Significantly, both aromatic and non-aromatic amino acids are detected and discriminated at the single-molecule level either at individual amino acid molecules or within the polypeptide chains. Correlated with molecular dynamics simulations, our results suggest that the signal dominance due to large spatial occupancy of aromatic rings of the polypeptide sidechains on gold surfaces can be overcome by the high localization of the single hot spot. The superior spectral and spatial discriminative power of our approach can be applied to single-protein analysis, fingerprinting, and sequencing.

SERS discrimination of single DNA bases in single oligonucleotides by electro-plasmonic trapping.

Surface-enhanced Raman spectroscopy (SERS) sensing of DNA bases by plasmonic nanopores could pave a way to novel methods for DNA analyses and new generation single-molecule sequencing platforms. The SERS discrimination of single DNA bases depends critically on the time that a DNA strand resides within the plasmonic hot spot. In fact, DNA molecules flow through the nanopores so rapidly that the SERS signals collected are not sufficient for single-molecule analysis. Here, we report an approach to control the residence time of molecules in the hot spot by an electro-plasmonic trapping effect. By directly adsorbing molecules onto a gold nanoparticle and then trapping the single nanoparticle in a plasmonic nanohole up to several minutes, we demonstrate single-molecule SERS detection of all four DNA bases as well as discrimination of single nucleobases in a single oligonucleotide. Our method can be extended easily to label-free sensing of single-molecule amino acids and proteins.

Comparative analysis of opto-electronic performance of aluminium and silver nano-porous and nano-wired layers.

The comparison of optical and electronic properties between squarely and hexagonally arranged nano-porous layers and uniformly arranged nano-wired layers of aluminium and silver was presented. The nano-wired configuration exhibit 20 and 10% higher average transmittance in visible wavelength range in comparison to square and hexagonal nano-porous designs, respectively. The insignificant difference of the transmittance for aluminium and silver nano-porous and nano-wired layers is observed, when interpore/interwire distance is larger than wavelengths of incoming light. This difference becomes considerable at the interpore/interwire distance less than wavelengths of incoming light: silver nano-porous and nano-wired layers possess up to 27% higher transmittance in comparison to aluminium layers.

Theoretical comparison of optical and electronic properties of uniformly and randomly arranged nano-porous ultra-thin layers.

The theoretical comparison of optical and electronic properties of aluminum and silver nano-porous ultra-thin layers in terms of the arrangement and size of the pores was presented. The uniform nano-porous layers exhibit a slightly higher average transmittance (up to 10%) in the wavelength range of the plasmonic response in comparison to the randomly arranged ones. Compared to uniform nano-porous layers, a much larger sheet resistance (up to 12 times) for random nano-porous layers is observed. The uniform and random Ag nano-porous layers possessing the strong plasmonic response over whole visible range can reach an average transmittance of 90 and 80% at the sheet resistance of 10 and 20 Ohm/sq, respectively, which is comparable to widely used ITO electrodes.

Ultra-thin broadband nanostructured insulator-metal-insulator-metal plasmonic light absorber.

An ultra-thin nanostructured plasmonic light absorber with an insulator-metal-insulator-metal (IMIM) architecture is designed and numerically studied. The IMIM structure is capable to absorb up to about 82.5% of visible light in a broad wavelength range of 300-750 nm. The absorption by the bottom metal is only 6% of that of the top metal. The results show that the IMIM architecture has weak dependence of the angle of the incident light. Interestingly, by varying the top insulator material the optical absorption spectrum can be shifted more than 180 nm as compared to the conventional air-metal-insulator-metal structure. The IMIM structure can be applied for different plasmonic devices with improved performance.

Optoelectronic performance optimization for transparent conductive layers based on randomly arranged silver nanorods.

Optoelectronic performance of transparent conductive layers (TCLs) based on randomly arranged silver (Ag) nanorods (NRs) is simulated. Models for calculation of optical and electronic properties were proposed founded on finite-difference time-domain method and percolation theory respectively. Obtained simulation results are well conformed to experimental data. The influence of angle deviation of NR crossings on the transmittance and sheet resistance are demonstrated. The balance between transmittance and sheet resistance which can be easily set by varying the combinations of NR radius and NR number is shown. Our results demonstrate that randomly arranged Ag layers are promising candidates for flexible TCLs.