A versatile technology for colloidal crystal transfer using parylene coatings and hydrosoluble polymers.

We propose a novel versatile colloidal crystal transfer technique compatible with a wide range of water-insoluble substrates regardless of their size, material, and wettability. There are no inherent limitations on colloidal particles material and size. The method possibilities are demonstrated via the colloidal transfer on quartz, glass substrates with a flat and curved surface, and via the fabrication of 3D colloidal structure with 5 overlaid colloidal monolayers. The process occurs at a room temperature in water and is independent from the illumination conditions, which makes it ideal for experimental manipulations with sensitive functional substrates. We performed the nanosphere photolithography process on a photosensitive substrate with a transferred colloidal monolayer. The metallized hexagonal arrays of nanopores demonstrated a clear resonant plasmonic behavior. We believe that due to its high integration possibilities the proposed transfer technique will find applications in a large-area surface nanotexturing, plasmonics, and will speed up a device fabrication process.

Laser induced mechanisms controlling the size distribution of metallic nanoparticles.

This paper describes a model to simulate changes in the size distribution of metallic nanoparticles (NPs) in TiO2 films upon continuous wave light excitation. Interrelated laser induced physical and chemical processes initiated directly by photon absorption or by plasmon induced thermal heating are considered. Namely the model takes into account the NP coalescence, Ostwald ripening, the reduction of silver ions and the oxidation of metallic NPs, competitive mechanisms that can lead to counter-intuitive behaviors depending on the exposure conditions. Theoretical predictions are compared successfully to the experimental results deduced from a thorough analysis of scanning transmission electron microscopy (STEM) pictures of Ag:TiO2 films processed with a scanning visible laser beam at different speeds. Ag:TiO2 systems are considered for many applications in solar energy conversion, photocatalysis or secured data printing. Numerical investigations of such a system provide a better understanding of light induced growth and shrinking processes and open up prospects for designing more efficient photocatalytic devices based on metal NP doped TiO2 or for improving the size homogeneity in self-organized metallic NP patterns, for instance.

Hybridization between plasmonic and photonic modes in laser-induced self-organized quasi-random plasmonic metasurfaces.

Plasmonic metasurfaces made of perfectly regular 2D lattices of metallic nanoparticles deposited on surfaces or close to waveguides can exhibit hybridized plasmonic and photonic modes. The latter arise from the excitation of surface or guided modes through the in-plane coherent scattering of periodic arrays. Recently, laser-induced self-organization of random plasmonic metasurfaces has been used to create nanoparticle gratings embedded in protective layers. Despite the broad size distribution and positional disorder of nanoparticles, the resulting nanostructures exhibit strong coupling between plasmonic and photonic modes in transverse electric polarization, leading to dichroism, which is well-reproduced from one laser printing to another. Here, we examine quantitatively the effect of inhomogeneities at the nanoscale on the hybridization between localized plasmonic modes and delocalized guided modes by considering realistic laser-induced self-organized nanoparticle arrays embedded in a two-layer system. By referring to regular samples, we describe the optical mechanisms involved in the hybridization process at characteristic wavelengths, based on far and near field simulations. Two kinds of real samples are considered, featuring different levels of coupling between the plasmonic and photonic modes. The results demonstrate that controlling the statistical properties of plasmonic metasurfaces, such as the nanoparticle size distribution and average position, over areas a few micrometers wide is enough to control in a reproducible manner the hybridization mechanisms and their resulting optical properties. Thus, this study shows that the inherent irregularities of laser-induced self-organized nanostructures are compatible with smart functionalities of nanophotonics, and confirms that laser processing has huge potential for real-world applications.

High-Density Nanowells Formation in Ultrafast Laser-Irradiated Thin Film Metallic Glass.

We present an effective approach for fabricating nanowell arrays in a one-step laser process with promising applications for the storage and detection of chemical or biological elements. Biocompatible thin films of metallic glasses are manufactured with a selected composition of Zr65Cu35, known to exhibit remarkable mechanical properties and glass forming ability. Dense nanowell arrays spontaneously form in the ultrafast laser irradiation spot with dimensions down to 20 nm. The flared shape observed by transmission electron microscopy is ideal to ensure chemical or biological material immobilization into the nanowells. This also indicates that the localization of the cavitation-induced nanopores can be tuned by the density and size of the initial nanometric interstice from the columnar structure of films deposited by magnetron sputtering. In addition to the topographic functionalization, the laser-irradiated amorphous material exhibits structural changes analyzed by spectroscopic techniques at the nanoscale such as energy-dispersive X-ray spectroscopy and electron energy loss spectroscopy. Results reveal structural changes consisting of nanocrystals of monoclinic zirconia that grow within the amorphous matrix. The mechanism is driven by local oxidation process catalyzed by extreme temperature and pressure conditions estimated by an atomistic simulation of the laser-induced nanowell formation.

Ent3p Is a PtdIns(3,5)P2 effector required for protein sorting to the multivesicular body.

PtdIns(3,5)P(2) is required for cargo-selective sorting to the vacuolar lumen via the multivesicular body (MVB). Here we show that Ent3p, a yeast epsin N-terminal homology (ENTH) domain-containing protein, is a specific PtdIns(3,5)P(2) effector localized to endosomes. The ENTH domain of Ent3p is essential for its PtdIns(3,5)P(2) binding activity and for its membrane interaction in vitro and in vivo. Ent3p is required for protein sorting into the MVB but not for the internalization step of endocytosis. Ent3p is associated with clathrin and is necessary for normal actin cytoskeleton organization. Our results show that Ent3p is required for protein sorting into intralumenal vesicles of the MVB through PtdIns(3,5)P(2) binding via its ENTH domain.

Involvement of the AP-1 adaptor complex in early steps of phagocytosis and macropinocytosis.

The best described function of the adaptor complex-1 (AP-1) is to participate in the budding of clathrin-coated vesicles from the trans-Golgi network and endosomes. Here, we show that AP-1 is also localized to phagocytic cups in murine macrophages as well as in Dictyostelium amoebae. AP-1 is recruited to phagosomal membranes at this early stage of phagosome formation and rapidly dissociates from maturing phagosomes. To establish the role of AP-1 in phagocytosis, we made used of Dictyostelium mutant cells (apm1(-) cells) disrupted for AP-1 medium chain. In this mutant, phagocytosis drops by 60%, indicating that AP-1 is necessary for efficient phagocytosis. Furthermore, phagocytosis in apm1(-) cells is more affected for large rather than small particles, and cells exhibiting incomplete engulfment are then often observed. This suggests that AP-1 could participate in the extension of the phagocytic cup. Interestingly, macropinocytosis, a process dedicated to fluid-phase endocytosis and related to phagocytosis, is also impaired in apm1(-) cells. In summary, our data suggest a new role of AP-1 at an early stage of phagosome and macropinosome formation.

Phg2, a kinase involved in adhesion and focal site modeling in Dictyostelium.

The amoeba Dictyostelium is a simple genetic system for analyzing substrate adhesion, motility and phagocytosis. A new adhesion-defective mutant named phg2 was isolated in this system, and PHG2 encodes a novel serine/threonine kinase with a ras-binding domain. We compared the phenotype of phg2 null cells to other previously isolated adhesion mutants to evaluate the specific role of each gene product. Phg1, Phg2, myosin VII, and talin all play similar roles in cellular adhesion. Like myosin VII and talin, Phg2 also is involved in the organization of the actin cytoskeleton. In addition, phg2 mutant cells have defects in the organization of the actin cytoskeleton at the cell-substrate interface, and in cell motility. Because these last two defects are not seen in phg1, myoVII, or talin mutants, this suggests a specific role for Phg2 in the control of local actin polymerization/depolymerization. This study establishes a functional hierarchy in the roles of Phg1, Phg2, myosinVII, and talin in cellular adhesion, actin cytoskeleton organization, and motility.

The AP-1 clathrin-adaptor is required for lysosomal enzymes sorting and biogenesis of the contractile vacuole complex in Dictyostelium cells.

Adaptor protein complexes (AP) are major components of the cytoplasmic coat found on clathrin-coated vesicles. Here, we report the molecular and functional characterization of Dictyostelium clathrin-associated AP-1 complex, which in mammalian cells, participates mainly in budding of clathrin-coated vesicles from the trans-Golgi network (TGN). The gamma-adaptin AP-1 subunit was cloned and shown to belong to a Golgi-localized 300-kDa protein complex. Time-lapse analysis of cells expressing gamma-adaptin tagged with the green-fluorescent protein demonstrates the dynamics of AP-1-coated structures leaving the Golgi apparatus and rarely moving toward the TGN. Targeted disruption of the AP-1 medium chain results in viable cells displaying a severe growth defect and a delayed developmental cycle compared with parental cells. Lysosomal enzymes are constitutively secreted as precursors, suggesting that protein transport between the TGN and lysosomes is defective. Although endocytic protein markers are correctly localized to endosomal compartments, morphological and ultrastructural studies reveal the absence of large endosomal vacuoles and an increased number of small vacuoles. In addition, the function of the contractile vacuole complex (CV), an osmoregulatory organelle is impaired and some CV components are not correctly targeted.

Spectral and Color Changes of Ag/TiO2 Photochromic Films Deposited on Diffusing Paper and Transparent Flexible Plastic Substrates.

Giving paper and polymer photochromic properties under laser irradiation is challenging due to the low resistance of these materials to heat, their flexibility, and their possibly irregular structure. However, we could successfully deposit TiO2/Ag/TiO2 layers stacking on flexible white glossy paper and transparent polyethylene terephalate (PET) substrates using a reactive magnetron sputtering technique, and tailor coloration changes after laser irradiation, alternating visible and ultraviolet (UV) wavelengths. The sample colors are characterized by a panel of chromas depending on the irradiation conditions. We demonstrate that these chroma changes are due to morphological changes of Ag nanoparticles (NPs) after visible laser irradiation of the colored as-deposited sample. The process exhibits a good reversibility after subsequent UV irradiation due to the growth of new metallic Ag NPs. The colors displayed in diffuse reflection by the paper samples are more saturated than the ones displayed in regular transmission by PET samples. We demonstrate the efficiency of the photochromic process on such support by printing high resolution patterns exhibiting different colors depending on the observation conditions.

Three-Dimensional Self-Organization in Nanocomposite Layered Systems by Ultrafast Laser Pulses.

Controlling plasmonic systems with nanometer resolution in transparent films and their colors over large nonplanar areas is a key issue for spreading their use in various industrial fields. Using light to direct self-organization mechanisms provides high-speed and flexible processes to meet this challenge. Here, we describe a route for the laser-induced self-organization of metallic nanostructures in 3D. Going beyond the production of planar nanopatterns, we demonstrate that ultrafast laser-induced excitation combined with nonlinear feedback mechanisms in a nanocomposite thin film can lead to 3D self-organized nanostructured films. The process, which can be extended to complex layered composite systems, produces highly uniform large-area nanopatterns. We show that 3D self-organization originates from the simultaneous excitation of independent optical modes at different depths in the film and is activated by the plasmon-induced charge separation and thermally induced NP growth mechanisms. This laser color marking technique enables multiplexed optical image encoding and the generated nanostructured Ag NPs:TiO2 films offer great promise for applications in solar energy harvesting, photocatalysis, or photochromic devices.

Microstructuring of Mesoporous Titania Films Loaded with Silver Salts to Enhance the Photocatalytic Degradation of Methyl Blue under Visible Light.

The microstructuring of the distribution of silver nanoparticles (NPs) in mesoporous titania films loaded with silver salts, using two-beam interference lithography leading to 1 Dimension (1D) grating, induces variations in the photocatalytic efficiency. The influence of the structuration was tested on the degradation of methyl blue (MB) under ultraviolet (UV) and visible illumination, giving rise to a significant improvement of the photocatalytic efficiency. The periodic distribution of the NPs was characterized by transmission electron microscopy (TEM), high-angle annular dark field scanning transmission electron microscopy (HAADF-STEM) and scanning electron microscopy (SEM).

A novel phosphatidylinositol 4,5-bisphosphate-binding domain targeting the Phg2 kinase to the membrane in Dictyostelium cells.

Phg2 is a ser/thr kinase involved in adhesion, motility, actin cytoskeleton dynamics, and phagocytosis in Dictyostelium cells. In a search for Phg2 domains required for its localization to the plasma membrane, we identified a new domain interacting with phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)) and phosphatidylinositol 4-phosphate (PI(4)P) membrane phosphoinositides. Deletion of this domain prevented membrane recruitment of Phg2 and proper function of the protein in the phagocytic process. Moreover, the overexpression of this PI(4,5)P(2)-binding domain specifically had a dominant-negative effect by inhibiting phagocytosis. Therefore, plasma membrane recruitment of Phg2 is essential for its function. The PI(4,5)P(2)-binding domain fused to GFP (green fluorescent protein) (GFP-Nt-Phg2) was also used to monitor the dynamics of PI(4,5)P(2) during macropinocytosis and phagocytosis. GFP-Nt-Phg2 disappeared from macropinosomes immediately after their closure. During phagocytosis, PI(4,5)P(2) disappeared even before the sealing of phagosomes as it was already observed in mammalian cells. Together these results demonstrate that PI(4,5)P(2) metabolism regulates the dynamics and the function of Phg2.

Acidic clusters target transmembrane proteins to the contractile vacuole in Dictyostelium cells.

The mechanisms responsible for the targeting of transmembrane integral proteins to the contractile vacuole (CV) network in Dictyostelium discoideum are unknown. Here we show that the transfer of the cytoplasmic domain of a CV-resident protein (Rh50) to a reporter transmembrane protein (CsA) is sufficient to address the chimera (CsA-Rh50) to the CV. We identified two clusters of acidic residues responsible for this targeting, and these motifs interacted with the gamma-adaptin AP-1 subunit in a yeast protein-protein interaction assay. For the first time we report the existence of an indirect transport pathway from the plasma membrane to the CV via endosomes. Upon internalization, the small fraction of CsA-Rh50 present at the cell surface was first concentrated in endosomes distinct from early and late p80-positive endosomes and then slowly transported to the CV. Together our results suggest the existence of an AP-1-dependent selective transport to the contractile vacuole in Dictyostelium.

Two members of the beige/CHS (BEACH) family are involved at different stages in the organization of the endocytic pathway in Dictyostelium.

Proteins of the Chediak-Higashi/Beige (BEACH) family have been implicated in the function of lysosomes, as well as in signal transduction, but their molecular role is still poorly understood. In Dictyostelium, at least six members of the family can be identified. Here cells with mutations in two of these genes, LVSA and LVSB, were analyzed. Interestingly both mutants exhibited defects in the organization of the endocytic pathway, albeit at distinct stages. In lvsB mutant cells, the regulated secretion of lysosomal enzymes was enhanced, a phenotype reminiscent of the Chediak-Higashi syndrome. LvsA mutant cells exhibited alterations in the organization and function of the early endocytic and phagocytic pathway. The LvsA protein may participate in the signaling pathway, which links adhesion of a particle to the subsequent formation of a phagocytic cup. Further genetic analysis will be necessary to determine whether other members of the BEACH family of proteins are also involved in controlling the organization of the endocytic pathway.