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Abstract Optical stimulation and control of muscle cell contraction opens up a number of interesting applications in hybrid robotic and medicine. Here we show that recently designed molecular phototransducer can be used to stimulate C2C12 skeletal muscle cells, properly grown to exhibit collective behaviour. C2C12 is a skeletal muscle cell line that does not require animal sacrifice Furthermore, it is an ideal cell model for evaluating the phototransducer pacing ability due to its negligible spontaneous activity. We study the stimulation process and analyse the distribution of responses in multinuclear cells, in particular looking at the consistency between stimulus and contraction. Contractions are detected by using an imaging software for object recognition. We find a deterministic response to light stimuli, yet with a certain distribution of erratic behaviour that is quantified and correlated to light intensity or stimulation frequency. Finally, we compare our optical stimulation with electrical stimulation showing advantages of the optical approach, like the reduced cell stress.

In this article we investigate a mathematical model for a retinal prosthesis made of organic polymer nanoparticles (NP) in the stationary regime. The model consists of a Drift-Diffusion system to describe free charge transport in the NP bulk; a Poisson-Nernst-Planck system to describe ion electrodiffusion in the solution surrounding the NP; and nonlinear transmission conditions at the NP-solution interface. To solve the model we use an iteration procedure for which we prove the existence and briefly comment the uniqueness of a fixed point under suitable smallness assumptions on model parameters. For system discretization we use a stabilized finite element method to prevent unphysical oscillations in the electric potential, carrier number densities and ion molar densities. Model predictions describe the amount of active chemical molecule accumulating at the neuron surface and highlight electrostatic effects induced by the sole presence of the nanoparticle. These results support the use of mathematical modeling as a virtual laboratory for the optimal design of bio-hybrid systems, whose investigation may be impervious due to experimental limits.

Measuring the transit time of a cell forced through a bottleneck is one of the most widely used techniques for the study of cell deformability in flow. It in turn provides an accessible and rapid way of obtaining crucial information regarding cell physiology. Many techniques are currently being investigated to reliably retrieve this time, but their translation to diagnostic-oriented devices is often hampered by their complexity, lack of robustness, and the bulky external equipment required. Herein, we demonstrate the benefits of coupling microfluidics with an optical method, like photocells, to measure the transit time. We exploit the femtosecond laser irradiation followed by chemical etching (FLICE) fabrication technique to build a monolithic 3D device capable of detecting cells flowing through a 3D non-deformable constriction which is fully buried in a fused silica substrate. We validated our chip by measuring the transit times of pristine breast cancer cells (MCF-7) and MCF-7 cells treated with Latrunculin A, a drug typically used to increase their deformability. A difference in transit times can be assessed without the need for complex external instrumentation and/or demanding computational efforts. The high throughput (4000–10,000 cells/min), ease of use, and clogging-free operation of our device bring this approach much closer to real scenarios.

Abstract Doped organic semiconductors play a central role in the development of several innovative optoelectronic and energy harvesting applications. Currently, the realization of thermoelectric generators, which require both hole‐ and electron‐transporting materials with high electrical conductivity, is strongly hindered by the scarce availability of stable solution‐processable n‐dopants and their limited efficiency. Herein, the synthesis of 4‐(1,3‐dimethyl‐2,3‐dihydro‐1H‐benzimidazol‐2‐yl)‐dibenzazepine (IStBI), a novel derivative belonging to the well‐known family of the benzimidazoline compounds, is presented. The functionalization with the planarized and rigid iminostilbene substituent allows, without significantly affecting the compound electronic structure, an efficient intercalation of the dopant molecules inside the ordered regions of thin films of the benchmark n‐type polymer poly(N,N′‐bis‐2‐octyldodecylnaphthalene‐1,4,5,8‐bis‐dicarboximide‐2,6‐diyl‐alt‐5,5′‐2,2′‐bithiophene) P(NDI2OD‐T2). Consequently, a maximum electrical conductivity of (1.14 ± 0.13) × 10−2 S cm−1 is recorded, exceeding by one order of magnitude what previously achieved upon solution doping of the reference P(NDI2OD‐T2) with benzimidazoline derivatives. The thermoelectric power factor is also simultaneously increased. The findings confirm that tailoring of the dopant chemical structure to improve structural interactions with the host semiconductors can be employed as a successful strategy to achieve more effective n‐doping, helping to close the performance gap with p‐type materials.

Abstract Recent studies have shown that bacterial membrane potential is dynamic and plays signaling roles. Yet, little is still known about the mechanisms of membrane potential dynamics regulation—owing to a scarcity of appropriate research tools. Optical modulation of bacterial membrane potential could fill this gap and provide a new approach for studying and controlling bacterial physiology and electrical signaling. Here, the authors show that a membrane‐targeted azobenzene (Ziapin2) can be used to photo‐modulate the membrane potential in cells of the Gram‐positive bacterium Bacillus subtilis. It is found that upon exposure to blue–green light (λ = 470 nm), isomerization of Ziapin2 in the bacteria membrane induces hyperpolarization of the potential. To investigate the origin of this phenomenon, ion‐channel‐deletion strains and ion channel blockers are examined. The authors found that in presence of the chloride channel blocker idanyloxyacetic acid‐94 (IAA‐94) or in absence of KtrAB potassium transporter, the hyperpolarization response is attenuated. These results reveal that the Ziapin2 isomerization can induce ion channel opening in the bacterial membrane and suggest that Ziapin2 can be used for studying and controlling bacterial electrical signaling. This new optical tool could contribute to better understand various microbial phenomena, such as biofilm electric signaling and antimicrobial resistance.

Summary: Non-genetic photostimulation is a novel and rapidly growing multidisciplinary field that aims to induce light-sensitivity in living systems by exploiting exogeneous phototransducers. Here, we propose an intramembrane photoswitch, based on an azobenzene derivative (Ziapin2), for optical pacing of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The light-mediated stimulation process has been studied by applying several techniques to detect the effect on the cell properties. In particular, we recorded changes in membrane capacitance, in membrane potential (Vm), and modulation of intracellular Ca2+ dynamics. Finally, cell contractility was analyzed using a custom MATLAB algorithm. Photostimulation of intramembrane Ziapin2 causes a transient Vm hyperpolarization followed by a delayed depolarization and action potential firing. The observed initial electrical modulation nicely correlates with changes in Ca2+ dynamics and contraction rate. This work represents the proof of principle that Ziapin2 can modulate electrical activity and contractility in hiPSC-CMs, opening up a future development in cardiac physiology.

Statistical analysis of the properties of single microparticles, such as cells, bacteria or plastic slivers, has attracted increasing interest in recent years. In this regard, field flow cytometry is considered the gold standard technique, but commercially available instruments are bulky, expensive, and not suitable for use in point-of-care (PoC) testing. Microfluidic flow cytometers, on the other hand, are small, cheap and can be used for on-site analyses. However, in order to detect small particles, they require complex geometries and the aid of external optical components. To overcome these limitations, here, we present an opto-fluidic flow cytometer with an integrated 3D in-plane spherical mirror for enhanced optical signal collection. As a result, the signal-to-noise ratio is increased by a factor of six, enabling the detection of particle sizes down to 1.5 µm. The proposed optofluidic detection scheme enables the simultaneous collection of particle fluorescence and scattering using a single optical fiber, which is crucial to easily distinguishing particle populations with different optical properties. The devices have been fully characterized using fluorescent polystyrene beads of different sizes. As a proof of concept for potential real-world applications, signals from fluorescent HEK cells and Escherichia coli bacteria were analyzed.

Abstract 2D metal monochalcogenides have recently attracted interest for photoelectrochemical (PEC) applications in aqueous electrolytes. Their optical bandgap in the visible and near‐infrared spectral region is adequate for energy conversion and photodetection/sensing. Their large surface‐to‐volume ratio guarantees that the charge carriers are photogenerated at the material/electrolyte interface, where redox reactions occur, minimizing recombination processes. However, solution‐processed photoelectrodes based on these materials exhibit energy conversion efficiencies that are far from the current state of the art expressed by established technologies. This work reports a systematic morphological, spectroscopic, and PEC characterization of solution‐processed films of photoactive InSe flakes for PEC‐type photodetectors. By optimizing the thickness and hybridizing InSe flakes with electrically conductive Sn:In2O3 (ITO) nanocrystals, photoanodes with a significant photoanodic response in both acidic and alkaline media are designed, reaching responsivity up to 60.0 mA W−1 (external quantum efficiency = 16.4%) at +0.4 V versus RHE under visible illumination. In addition, a strategy based on the use of sacrificial agents (i.e., 2‐propanol and Na2SO3) is proposed to improve the stability of the InSe and ITO/InSe photodetectors. Our data confirm the potential of 2D InSe for PEC energy conversion and sensing applications, remarking the challenges related to InSe stability during anodic operation.

Reducing the thickness of flexible electronics is crucial to achieve better conformability and mechanical flexibility. Here, the authors demonstrate all-solution processed OFET with total thickness < 150 nm, with high transparency, which allows the device to conformally adhere onto non-planar surfaces, such as human skin.

The desire to increase the performance of optoelectronic devices has driven research into new, versatile optical materials. Among these, materials combinations composed of organic polymers and inorganic materials have sparked a strong interest due to the wide range of refractive indices that are obtainable. However, in parallel to these materials design and discovery activities, simple, broadly applicable fabrication techniques need to be advanced that are adapted to novel materials systems and allow, for instance, straightforward, large‐area manufacturing and integration of 3D photonic circuits. Herein, the use of a 3D direct writing laser technique is proposed to locally enhance the refractive index of a recently developed inorganic/organic molecular hybrid that is only composed of the commodity polymer poly(vinyl alcohol) cross‐linked with titanium oxide hydrates, for the creation of photonic structures. A spatially controlled refractive index increase of 0.13 between pristine and laser‐treated regions is achieved, with the potential of this adaptable 3D‐fabrication technique being demonstrated by manufacturing an optical diffraction grating. Considering that the synthesis and deposition of the hybrid material is water based and does not require toxic heavy metals, the work, thus, provides a processing and materials platform toward environment‐friendly, green processing of integrated photonic systems and beyond.

Abstract Diatom microalgae have great industrial potential as next-generation sources of biomaterials and biofuels. Effective scale-up of their production can be pursued by enhancing the efficiency of their photosynthetic process in a way that increases the solar-to-biomass conversion yield. A proof-of-concept demonstration is given of the possibility of enhancing the light absorption of algae and of increasing their efficiency in photosynthesis by in vivo incorporation of an organic dye which acts as an antenna and enhances cells’ growth and biomass production without resorting to genetic modification. A molecular dye (Cy5) is incorporated in Thalassiosira weissflogii diatom cells by simply adding it to the culture medium and thus filling the orange gap that limits their absorption of sunlight. Cy5 enhances diatoms’ photosynthetic oxygen production and cell density by 49% and 40%, respectively. Cy5 incorporation also increases by 12% the algal lipid free fatty acid (FFA) production versus the pristine cell culture, thus representing a suitable way to enhance biofuel generation from algal species. Time-resolved spectroscopy reveals Förster Resonance Energy Transfer (FRET) from Cy5 to algal chlorophyll. The present approach lays the basis for non-genetic tailoring of diatoms’ spectral response to light harvesting, opening up new ways for their industrial valorization.

Non-genetic photostimulation, which allows for control over cellular activity via the use of cell-targeting phototransducers, is widely used nowadays to study and modulate/restore biological functions. This approach relies on non-covalent interactions between the phototransducer and the cell membrane, thus implying that cell conditions and membrane status can dictate the effectiveness of the method. For instance, although immortalized cell lines are traditionally used in photostimulation experiments, it has been demonstrated that the number of passages they undergo is correlated to the worsening of cell conditions. In principle, this could impact cell responsivity against exogenous stressors, including photostimulation. However, these aspects have usually been neglected in previous experiments. In this work, we investigated whether cell passages could affect membrane properties (such as polarity and fluidity). We applied optical spectroscopy and electrophysiological measurements in two different biological models: (i) an epithelial immortalized cell line (HEK-293T cells) and (ii) liposomes. Different numbers of cell passages were compared to a different morphology in the liposome membrane. We demonstrated that cell membranes show a significant decrease in ordered domains upon increasing the passage number. Furthermore, we observed that cell responsivity against external stressors is markedly different between aged and non-aged cells. Firstly, we noted that the thermal-disordering effect that is usually observed in membranes is more evident in aged cells than in non-aged ones. We then set up a photostimulation experiment by using a membrane-targeted azobenzene as a phototransducer (Ziapin2). As an example of a functional consequence of such a condition, we showed that the rate of isomerization of an intramembrane molecular transducer is significantly impaired in aged cells. The reduction in the photoisomerization rate translates in cells with a sustained reduction of the Ziapin2-related hyperpolarization of the membrane potential and an overall increase in the molecule fluorescence. Overall, our results suggest that membrane stimulation strongly depends on membrane order, highlighting the importance of cell passage during the characterization of the stimulation tools. This study can shine light on the correlation between aging and the development of diseases driven by membrane degradation as well as on the different cell responsivities against external stressors, such as temperature and photostimulation.

The biological functions of the cell membrane are regulated by its physicochemical properties, such as the transmembrane potential, phase state and hydration. Therefore, the in situ investigation of these properties is attracting a rising interest in the field of membrane biophysics. In these regards, a wide variety of fluorescent membrane probes has been developed, providing powerful tools for fluorescence microscopy and/or fluorescence spectroscopy to study the microscopic analogs of viscosity, polarity, and hydration, as well as the molecular order and electrostatic potential at the sites of their location. Despite the success of this approach, further developments rely on the design of fluorescence probes with optimized photophysical properties. This review discusses the most successful fluorescent molecular probes and their application to the study of the plasma membrane viscosity, lipid order and potential.

The use of organic semiconductor nanoparticles (NPs) as retinal prostheses is attracting attention due to the possibility of injecting them directly into the desired tissue, with a minimally invasive surgical treatment. Polythiophene NPs localize in close proximity to the bipolar cell plasma membrane, which engulfs them, creating an intimate contact between the NP and the neuron. The intimate contact coupled with NP photoactivity are hypothesized to be the main guarantors of the electrostatic functioning of the bio-hybrid device. Since they may both be strongly affected by the geometric features of the NP, in this work, we use mathematical modeling to study the electrostatic polarization induced by light onto the NP and analyze how its spatial distribution is modified by varying the radius of the NP and its shape. Simulation results support the efficacy of the theoretical approach as a complementary virtual laboratory in the optimization of the current device and in the development of similar future NP-based technologies.

Cell membrane perturbation is a common way to stimulate cells by using external actuators. Recently, nanotechnology has added a number of new strategies for doing this, enlarging the scope and the range of mechanisms involved. Here, we describe a number of possible perturbation actions that are driven by light, and we try to capture the underlying phenomena. The discussion is based on the simple equivalent circuit model for the cell membrane.

Substrate engineering for steering cell growth is a wide and well‐established area of research in the field of modern biotechnology. Here we introduce a micromachining technique to pattern an inert and transparent polymer matrix blended with a photoactive polymer. We demonstrate that the obtained scaffold combines the capability to align with that to photostimulate living cells. This technology can open up new and promising applications, especially where cell alignment is required to trigger specific biological functions, e.g., generate powerful and efficient muscle contractions following an external stimulus.

We present a series of cationic membrane-targeted azobenzene molecules, with the aim to understand how variations in molecular architecture influence the relative optical and biological properties. 1,4-Amino-substituted azobenzene was chosen as the switching unit while the number of linked alkyl chains and their cationic end-group were systematically varied. Their photophysics, membrane partitioning, and electrophysiological efficacy were studied. We found that the polar end group is a key-factor determining the interaction with the phospholipid heads in the plasma membrane bilayer and consequently the ability to dimerize. The monosubstituted photoswitch with a pyridinium-terminated alkyl chain was found to be the best candidate for photostimulation. This study provides a structure-property investigation that can guide the chemical engineering of a new generation of molecular photo-actuators.

Photonic crystals can be employed effectively as simple and low-cost colorimetric sensors for monitoring variation in the environmental refractive index. In most cases, the photonic colorimetric approach relies on the use of porous and permeable materials to highlight refractive index (RI) modulation, although a fine control over the size distribution and free volume can be complex to achieve. Here, we propose nonporous low-layer count distributed Bragg reflectors (DBRs) as simple optical devices for colorimetric refractive index sensing. In our feasibility study, we simulated the reflectance of DBRs consisting of two to five SiO2/TiO2 bilayers upon variation of the external refractive index. We found that the 2-bilayers sample exhibits the highest sensitivity to RI variations, and identified the ratio between the first and third order reflectance intensity as simple yet efficient ratiometric parameter to discern analytes with different refractive indices. This approach can provide a promising perspective for the development of cheap and portable devices for environmental detection of a wide range of substances.

The progressive degeneration of retinal photoreceptors is one of the most significant causes of blindness in humans. Conjugated polymers represent an attractive solution to the field of retinal prostheses, and a multi-layer fully organic prosthesis implanted subretinally in dystrophic Royal College of Surgeons (RCS) rats was able to rescue visual functions. As a step toward human translation, we report here the fabrication and in vivo testing of a similar device engineered to adapt to the human-like size of the eye of the domestic pig, an excellent animal paradigm to test therapeutic strategies for photoreceptors degeneration. The active conjugated polymers were layered onto two distinct passive substrates, namely electro-spun silk fibroin (ESF) and polyethylene terephthalate (PET). Naive pigs were implanted subretinally with the active device in one eye, while the contralateral eye was sham implanted with substrate only. Retinal morphology and functionality were assessed before and after surgery by means of in vivo optical coherence tomography and full-field electroretinogram (ff-ERG) analysis. After the sacrifice, the retina morphology and inflammatory markers were analyzed by immunohistochemistry of the excised retinas. Surprisingly, ESF-based prostheses caused a proliferative vitreoretinopathy with disappearance of the ff-ERG b-wave in the implanted eyes. In contrast, PET-based active devices did not evoke significant inflammatory responses. As expected, the subretinal implantation of both PET only and the PET-based prosthesis locally decreased the thickness of the outer nuclear layer due to local photoreceptor loss. However, while the implantation of the PET only substrate decreased the ff-ERG b-wave amplitude with respect to the pre-implant ERG, the eyes implanted with the active device fully preserved the ERG responses, indicating an active compensation of the surgery-induced photoreceptor loss. Our findings highlight the possibility of developing a new generation of conjugated polymer/PET-based prosthetic devices that are highly biocompatible and potentially suitable for subretinal implantation in patients suffering from degenerative blindness.

Real-time monitoring of bacterial contaminants and pollutants in food is of paramount importance nowadays, owing to the impressive extension of the food production/supply chain and the consequent increase in foodborne outbreaks worldwide. This represents a serious risk for consumers’ health and accounts for a large fraction of food wastage, especially in the developed countries. Therefore, modern sensors for food quality control should possibly afford low-cost, portability, and easiness of readout to enable widespread diffusion of the technology, thus allowing food quality monitoring from the production/supply chain to the consumers’ table. In these regards, one-dimensional photonic crystals, also known as Distributed Bragg Reflectors (DBRs), can represent simple yet efficient all-optical and label-free colorimetric sensors, given their relatively high color purity, easiness of integration with a large number of stimulus responsive materials, and low-cost fabrication from scalable processes. In this perspective article, we discuss the development of DBRs-based colorimetric sensors for the monitoring of bacterial contaminants and pollutants of interest in the food quality sector. We aim at providing a systematic overview on the main approaches that have been employed to achieve selectivity and sensitivity in DBRs-based sensors, with the view to enable widespread use of this technology at both the industry/supply chain and customers’ level.

Abstract The non‐covalent affinity of photoresponsive molecules to biotargets represents an attractive tool for achieving effective cell photo‐stimulation. Here, an amphiphilic azobenzene that preferentially dwells within the plasma membrane is studied. In particular, its isomerization dynamics in different media is investigated. It is found that in molecular aggregates formed in water, the isomerization reaction is hindered, while radiative deactivation is favored. However, once protected by a lipid shell, the photochromic molecule reacquires its ultrafast photoisomerization capacity. This behavior is explained considering collective excited states that may form in aggregates, locking the conformational dynamics and redistributing the oscillator strength. By applying the pump probe technique in different media, an isomerization time in the order of 10 ps is identified and the deactivation in the aggregate in water is also characterized. Finally, it is demonstrated that the reversible modulation of membrane potential of HEK293 cells via illumination with visible light can be indeed related to the recovered trans→cis photoreaction in lipid membrane. These data fully account for the recently reported experiments in neurons, showing that the amphiphilic azobenzenes, once partitioned in the cell membrane, are effective light actuators for the modification of the electrical state of the membrane.

Recently, tunable photonic crystals (PhCs) have received great research interest, thanks to the wide range of applications in which they can be employed, such as light emission and sensing, among others. In addition, the versatility and ease of fabrication of PhCs allow for the integration of a large range of responsive elements that, in turn, can permit active tuning of PhC optical properties upon application of external stimuli, e.g., physical, chemical or even biological triggers. In this work, we summarize the most employed theoretical tools used for the design of optical properties of responsive PhCs and the most used fabrication techniques. Furthermore, we collect the most relevant results related to this field, with particular emphasis on electrochromic devices.

Chemically synthesized zigzag-edged nanographenes (NG) have recently demonstrated great success as the active laser units in solution-processed organic distributed feedback (DFB) lasers. Here, we report the first observation of dual amplified spontaneous emission (ASE) from a large-size NG derivative (with 12 benzenoid rings) dispersed in a polystyrene film. ASE is observed simultaneously at the 685 and 739 nm wavelengths, which correspond to different transitions of the photoluminescence spectrum. Ultrafast pump-probe spectroscopy has been used to ascertain the underlying photophysical processes taking place in the films. DFB lasers, based on these materials and top-layer nanostructured polymeric resonators (i.e., one or two-dimensional surface relief gratings), have been fabricated and characterized. Lasers emitting close to either one of the two possible ASE wavelengths, or simultaneously at both of them, have been prepared by proper selection of the resonator parameters.

Optical modulation of living cells activity by light-absorbing exogenous materials is gaining increasing interest, due to the possibility both to achieve high spatial and temporal resolution with a minimally invasive and reversible technique and to avoid the need of viral transfection with light-sensitive proteins. In this context, conjugated polymers represent ideal candidates for photo-transduction, due to their excellent optoelectronic and biocompatibility properties. In this work, we demonstrate that organic polymer nanoparticles, based on poly(3-hexylthiophene) conjugated polymer, establish a functional interaction with an in vitro cell model (Human Embryonic Kidney cells, HEK-293). They display photocatalytic activity in aqueous environment and, once internalized within the cell cytosol, efficiently generate reactive oxygen species (ROS) upon visible light excitation, without affecting cell viability. Interestingly, light-activated ROS generation deterministically triggers modulation of intracellular calcium ion flux, successfully controlled at the single cell level. In perspective, the capability of polymer NPs to produce ROS and to modulate Ca2+ dynamics by illumination on-demand, at non-toxic levels, may open the path to the study of biological processes with a gene-less approach and unprecedented spatio-temporal resolution, as well as to the development of new biotechnology tools for cell optical modulation.

In recent years, there has been significant research on integrated microfluidic devices. Microfluidics offer an advantageous platform for the parallel laminar flow of adjacent solvents of potential use in modern chemistry and biology. To reach that aim, we worked towards the realization of a buried microfluidic Lab-on-a-Chip which enables the separation of the two components by exploiting the non-mixing properties of laminar flow. To fabricate the aforementioned chip, we employed a femtosecond laser irradiation technique followed by chemical etching. To optimize the configuration of the chip, several geometrical and structural parameters were taken into account. The diffusive mass transfer between the two fluids was estimated and the optimal chip configuration for low diffusion rate of the components was defined.