Cyclic olefin copolymer (COC) as a promising biomaterial for affecting bacterial colonization: Investigation on Vibrio campbellii.

Cyclic olefin copolymer (COC) has emerged as an interesting biocompatible material for Organ-on-a-Chip (OoC) devices monitoring growth, viability, and metabolism of cells. Despite ISO 10993 approval, systematic investigation of bacteria grown onto COC is a still not documented issue. This study discusses biofilm formations of the canonical wild type BB120 Vibrio campbellii strain on a native COC substrate and addresses the impact of the physico-chemical properties of COC compared to conventional hydroxyapatite (HA) and poly(dimethylsiloxane) (PDMS) surfaces. An interdisciplinary approach combining bacterial colony counting, light microscopy imaging and advanced digital image processing remarks interesting results. First, COC can reduce biomass adhesion with respect to common biopolymers, that is suitable for tuning biofilm formations in the biological and medical areas. Second, remarkably different biofilm morphology (dendritic complex patterns only in the case of COC) was observed among the examined substrates. Third, the observed biofilm morphogenesis was related to the interaction of COC with the conditioning layer of the planktonic biological medium. Fourth, Grey Level Co-occurrence Matrix (CGLM)-based analysis enabled quantitative assessment of the biomass textural fractal development under different coverage conditions. All of this is of key practical relevance in searching innovative biocompatible materials for pharmaceutical, implantable and medical products.

Polymeric Membranes Doped with Halloysite Nanotubes Imaged using Proton Microbeam Microscopy.

Polymeric membranes are useful tools for water filtration processes, with their performance strongly dependent on the presence of hydrophilic dopants. In this study, polyaniline (PANI)-capped aluminosilicate (halloysite) nanotubes (HNTs) are dispersed into polyether sulfone (PES), with concentrations ranging from 0.5 to 1.5 wt%, to modify the properties of the PES membrane. Both undoped and HNT-doped PES membranes are investigated in terms of wettability (static and time-dependent contact angle), permeance, mechanical resistance, and morphology (using scanning electron microscopy (SEM)). The higher water permeance observed for the PES membranes incorporating PANI-capped HNTs is, finally, assessed and discussed vis-à-vis the real distribution of HNTs. Indeed, the imaging and characterization in terms of composition, spatial arrangement, and counting of HNTs embedded within the polymeric matrix are demonstrated using non-destructive Micro Particle Induced X-ray Emission (µ-PIXE) and Scanning Transmission Ion Microscopy (STIM) techniques. This approach not only exhibits the unique ability to detect/highlight the distribution of HNTs incorporated throughout the whole thickness of polymer membranes and provide volumetric morphological information consistent with SEM imaging, but also overcomes the limits of the most common analytical techniques exploiting electron probes. These aspects are comprehensively discussed in terms of practical analysis advantages.

Cooperation in bioluminescence: understanding the role of autoinducers by a stochastic random resistor model.

Quorum sensing is a communication mechanism adopted by different bacterial strains for the regulation of gene transcription. It takes place through the exchange of molecules called autoinducers. Bioluminescence is an emergent threshold phenomenon shown by some bacteria strains. Its precise relationship to quorum sensing is a debated topic, particularly regarding the role of the different autoinducers used by bacteria. In this paper, assuming a direct relationship between bioluminescence and quorum sensing, we investigate the role of multiple autoinducers in the bioluminescence response of Vibrio harveyi, considered as a model bioluminescent strain, due to its quorum sensing circuitry involving an array of three different autoinducers. Experiments on mutants of this bacterium, obtained by suppression of one or more autoinducers, reveal their relative non-trivial relevance and cooperative interaction patterns. The proposed analysis is implemented on a regular lattice, whose nodes represent microbial entities equipped with charges, which represent the ability to up/down regulate the gene expression. Quorum sensing results from a Coulomb-type field, produced by the charges. In analogy with random resistor network models, the lattice is permeated by an effective current which accounts for the amount and distribution of the charges. We propose that the presence of different autoinducers correspond to a different up/down regulation of gene expression, i.e., to a different way to account for the charges. Then, by introducing a modulation of the charge dependence into the current flowing within the network, we show that it is able to describe the bioluminescence exhibited by V. harveyi mutants. Furthermore, modulation of the charge dependence allows the interactions between the different autoinducers to be taken into account, providing a prediction regarding the data obtainable under specific growth conditions.

Mutant-Dependent Local Orientational Correlation in Biofilms of Vibrio campbellii Revealed through Digital Processing of Light Microscopy Images.

Biofilms are key bacterial communities in genetic and adaptive resistance to antibiotics as well as disease control strategies. The mature high-coverage biofilm formations of the Vibrio campbellii strains (wild type BB120 and isogenic derivatives JAF633, KM387, and JMH603) are studied here through the unstraightforward digital processing of morphologically complex images without segmentation or the unrealistic simplifications used to artificially simulate low-density formations. The main results concern the specific mutant- and coverage-dependent short-range orientational correlation as well as the coherent development of biofilm growth pathways over the subdomains of the image. These findings are demonstrated to be unthinkable based only on a visual inspection of the samples or on methods such as Voronoi tessellation or correlation analyses. The presented approach is general, relies on measured rather than simulated low-density formations, and could be employed in the development of a highly efficient screening method for drugs or innovative materials.

MEED: A novel robust contrast enhancement procedure yielding highly-convergent thresholding of biofilm images.

Morphological and statistical investigation of biofilm images may be even more critical than the image acquisition itself, in particular in the presence of morphologically complex distributions, due to the unavoidable impact of the measurement technique too. Hence, digital image pre-processing is mandatory for reliable feature extraction and enhancement preliminary to segmentation. Also, pattern recognition in automated deep learning (both supervised and unsupervised) models often requires a preliminary effective contrast-enhancement. However, no universal consensus exists on the optimal contrast enhancement approach. This paper presents and discusses a new general, robust, reproducible, accurate and easy to implement contrast enhancement procedure, briefly named MEED-procedure, able to work on images with different bacterial coverages and biofilm structures, coming from different imaging instrumentations (herein stereomicroscope and transmission microscope). It exploits a proper succession of basic morphological operations (erosion and dilation) and a horizontal line structuring element, to minimize the impact on size and shape of the even finer bacterial features. It systematically enhances the objects of interest, without histogram stretching and/or undesirable artifacts yielded by common automated methods. The quality of the MEED-procedure is ascertained by segmentation tests which demonstrate its robustness regarding the determination of threshold and convergence of the thresholding algorithm. Extensive validation tests over a rich image database, comparison with the literature and comprehensive discussion of the conceptual background support the superiority of the MEED-procedure over the existing methods and demonstrate it is not a routine application of morphological operators.

Potential of CO2-laser processing of quartz for fast prototyping of microfluidic reactors and templates for 3D cell assembly over large scale.

Carbon dioxide (CO2)-laser processing of glasses is a versatile maskless writing technique to engrave micro-structures with flexible control on shape and size. In this study, we present the fabrication of hundreds of microns quartz micro-channels and micro-holes by pulsed CO2-laser ablation with a focus on the great potential of the technique in microfluidics and biomedical applications. After discussing the impact of the laser processing parameters on the design process, we illustrate specific applications. First, we demonstrate the use of a serpentine microfluidic reactor prepared by combining CO2-laser ablation and post-ablation wet etching to remove surface features stemming from laser-texturing that are undesirable for channel sealing. Then, cyclic olefin copolymer micro-pillars are fabricated using laser-processed micro-holes as molds with high detail replication. The hundreds of microns conical and square pyramidal shaped pillars are used as templates to drive 3D cell assembly. Human Umbilical Vein Endothelial Cells are found to assemble in a compact and wrapping way around the micro-pillars forming a tight junction network. These applications are interesting for both Lab-on-a-Chip and Organ-on-a-Chip devices.

Pulsed Laser Deposition of CsPbBr3 Films: Impact of the Composition of the Target and Mass Distribution in the Plasma Plume.

All-inorganic cesium lead bromine (CsPbBr3) perovskites have gained a tremendous potential in optoelectronics due to interesting photophysical properties and much better stability than the hybrid counterparts. Although pulsed laser deposition (PLD) is a promising alternative to solvent-based and/or thermal deposition approaches due to its versatility in depositing multi-elemental materials, deep understanding of the implications of both target composition and PLD mechanisms on the properties of CsPbBr3 films is still missing. In this paper, we deal with thermally assisted preparation of mechano-chemically synthesized CsPbBr3 ablation targets to grow CsPbBr3 films by PLD at the fluence 2 J/cm2. We study both Cs rich- and stoichiometric PbBr2-CsBr mixture-based ablation targets and point out compositional deviations of the associated films resulting from the mass distribution of the PLD-generated plasma plume. Contrary to the conventional meaning that PLD guarantees congruent elemental transfer from the target to the substrate, our study demonstrates cation off-stoichiometry of PLD-grown CsPbBr3 films depending on composition and thermal treatment of the ablation target. The implications of the observed enrichment in the heavier element (Pb) and deficiency in the lighter element (Br) of the PLD-grown films are discussed in terms of optical response and with the perspective of providing operative guidelines and future PLD-deposition strategies of inorganic perovskites.

Short-range ordered 2D nanoholes: lattice-model and novel insight into the impact of coordination geometry and packing on their propagating-mode transmittance features.

Optically thin perforated gold films, fabricated using template colloidal masks self-assembled by following an elsewhere described simplified colloidal lithography protocol, are presented and discussed with the aim to develop a theory of short-range ordered nanoholes without straightforwardly extending concepts strictly related to periodic nanoholes. By Scanning Electron Microscopy (SEM) analysis of the evolution of nanohole short-range ordering and spatial coordination geometry under increasing interhole average spacing (d NN), unprecedented differences in the spectroscopic response are pointed out with respect to periodic systems. First, the dependence of the wavelength of a propagating plasmon mode on d NN is demonstrated to deviate from the linear relationship predicted by the grating-coupling picture developed for periodic arrays. Second, d NN cannot be straightforwardly interpreted as the counterpart of the lattice constant of periodic nanoholes, which demands to introduce a conceptually more rigorous periodicity-like length-scale. Once the impact of these findings on setting the operating parameters of a nanohole distribution is demonstrated, they are related, experimentally and by using a theoretical model developed by the authors, to the changes of the local coordination geometry (from quasi-hexagonal to quasi-square packing through mixed hexagonal-square coordination) induced by varying d NN over a wide interval. Autocorrelation analysis of SEM images is exploited to estimate a short-range periodicity-like length-scale, as a conceptual advance for laying the foundation of the concept of short-range ordered nanohole lattices and for deeper insight into the spectral response. As discussion is based on realistic, rather than simulated, evolution of colloidal arrangements, the formulated interpretative model accounts for realistic effects impacting transmission resonances.

Nanophosphors-Based White Light Sources.

Miniaturization requests and progress in nanofabrication are prompting worldwide interest in nanophosphors as white-emission mercury-free lighting sources. By comparison with their bulk counterparts, nanophosphors exhibit reduced concentration quenching effects and a great potential to enhance luminescence efficiency and tunability. In this paper, the physics of the nanophoshors is overviewed with a focus on the impact of spatial confinement and surface-to-volume ratio on the luminescence issue, as well as rare earth-activated multicolor emission for white light (WL) output. In this respect, the prominently practiced strategies to achieve WL emission are single nanophosphors directly yielding WL by means of co-doping and superposition of the individual red, green, and blue emissions from different nanophosphors. Recently, a new class of efficient broadband WL emitting nanophosphors has been proposed, i.e., nominally un-doped rare earth free oxide (yttrium oxide, Y2O3) nanopowders and Cr transition metal-doped garnet nanocrystals. In regard to this unconventional WL emission, the main points are: it is strictly a nanoscale phenomenon, the presence of an emitting center may favor WL emission without being necessary for observing it, and, its inherent origin is still unknown. A comparison between such an unconventional WL emission and the existing literature is presented to point out its novelty and superior lighting performances.

Gold nanoholes fabricated by colloidal lithography: novel insights into nanofabrication, short-range correlation and optical properties.

Colloidal lithography is widely used as a low cost and large-area deposition approach, alternative to the conventional small-area expensive lithographic techniques, for the fabrication of short-range ordered sub-wavelength metallic nanostructures. This paper contributes to the understanding of the impact of the fabrication protocol of a colloidal mask on the optical and sensing properties of short range-ordered nanohole (NH) distributions fabricated by colloidal lithography in optically thin (20 nm thick) gold films. We consider polystyrene nanospheres (PS-NSPs) with a nominal diameter of 80 nm, electrostatically adsorbed from a salt-free colloidal solution onto a polydiallyldimethylammonium (PDDA) countercharged monolayer. By avoiding the conventional polyelectrolyte multilayer and based on the interplay between the deposition times of both PDDA and PS-NSPs, we demonstrate effective simplification of the commonly applied deposition protocol and effective tuning of the NH-to-NH spacing (dNN) with negligible agglomeration. Comparison with NH samples prepared by salt-containing colloidal solutions points out the negative impact of salt addition on the optical properties. The effective tuning of dNN obtained by our protocol demonstrates highly correlated disorder under unsaturated adsorption and allows a discussion on the analogies of the optical response between long- and short- range ordered NH systems, which is a still debated topic. By Fast Fourier Transform of autocorrelation images of scanning electron microscopy micrographs we demonstrate quantitatively, rather than in principle, the correspondence between an inherent ordering length-scale and dNN. As optical transducers for detecting refractive index changes, our samples exhibit significant bulk sensitivity (∼309 nm RIU-1) in the framework of short range ordered NH systems.