Directed assembly of gold nanowires on silicon via reorganization and simultaneous fusion of randomly distributed gold nanoparticles.

Laser-induced reorganization and simultaneous fusion of nanoparticles is introduced as a versatile concept for pattern formation on surfaces. The process takes advantage of a phenomenon called laser-induced periodic surface structures (LIPSS) which originates from periodically alternating photonic fringe patterns in the near-field of solids. Associated photonic fringe patterns are shown to reorganize randomly distributed gold nanoparticles on a silicon wafer into periodic gold nanostructures. Concomitant melting due to optical heating facilitates the formation of continuous structures such as periodic gold nanowire arrays. Generated patterns can be converted into secondary structures using directed assembly or self-organization. This includes for example the rotation of gold nanowire arrays by arbitrary angles or their fragmentation into arrays of aligned gold nanoparticles.

Assembling Carbon Nanotube Architectures.

Well-defined multiwalled carbon nanotube structures are generated on stainless steel AISI 304 (EN AW 1.4301) by chemical vapor deposition. Pulsed laser-induced dewetting (PLiD) of the surface, by 532 nm nanosecond laser pulses, is utilized for the preparation of metal oxide nanoparticle fields with a defined particle number per area. The reduction of the precursor particles is achieved in an Ar/H2 (10% H2) atmosphere at 750 °C, thereby generating catalytic nanoparticles (c-NPs) for carbon nanotube (CNT) growth. Ethylene is used as a precursor gas for CNT growth. CNT lengths and morphology are directly related to the c-NP aerial density, which is dependent on the number of dewetting cycles during the PLiD process. Within a narrow window of c-NP per area, vertically aligned carbon nanotubes of great lengths are obtained. For more intense laser treatments, three-dimensional dewetting occurs and results in the formation of cauliflower-like structures. The laser process enables the creation of all kinds of CNT morphologies nearby on the microscale.

Formation of Highly Ordered Platinum Nanowire Arrays on Silicon via Laser-Induced Self-Organization.

Laser-induced periodic surface structures (LIPSS) provide an elegant solution for the generation of highly ordered periodic patterns on the surface of solids. In this study, LIPSS are utilized for the formation of periodic platinum nanowire arrays. In a process based on laser-stimulated self-organization, platinum thin films, sputter-deposited onto silicon, are transformed into nanowire arrays with an average periodicity of 538 nm. The width of the platinum nanowires is adjustable in a range from 20 nm to 250 nm by simply adjusting the thickness of the initial platinum thin films in a range from 0.3 nm to 4.3 nm. With increasing width, platinum nanowires show a rising tendency to sink into the surface of the silicon wafer, thus indicating alloying between platinum and silicon upon LIPSS-formation by a nanosecond-pulsed laser. The Pt/silicon wires may be etched away, leaving a complementary nanostructure in the silicon surface.

Light- and pH-dependent conformational changes in protein structure induce strong bending of purple membranes--active membranes studied by cryo-SEM.

Bacteriorhodopsin (BR) undergoes a conformational change during the photocycle and the proton transport through the membrane. For the first time, we could demonstrate by direct imaging of freely suspended native purple membranes (PMs) that the flat disk-like shape of PMs changes dramatically as soon as most of the BRs are in a state characterized by a deprotonated Schiff base. Light-induced shape changes are easily observed with mutated BRs of the BR-D96N type, i.e., all variants which show an increased M 2 lifetime. On the other hand, large-scale shape changes are induced by pH changes with PM containing mutated BRs of the BR-D85T type, where Asp85 is replaced for a neutral amino acid. In such PMs, all BRs are titrated simultaneously and the resulting shape of the membranes depends on the initial shape only. As the majority of PMs in the "flat" state are more or less round disks, the bent membranes often comprise bowl-like and tube-like bent structures. The method presented here enables one to derive size changes of membrane-embedded BRs on the single molecule level from "macroscopic", easily accessible data like the curvature radii observed in cryo-SEM. The potential of BR as a pH-controlled and/or light-controlled microscaled biological actuator needs further consideration.

Sugar-induced blue membrane: release of divalent cations during phase transition of purple membranes observed in sugar-derived glasses.

The formation of blue membrane from purple membranes (PM) has been observed in glassy films made from PM and various sugars. The phase transition of PM at about 70 degrees C causes the complexation of divalent cations to be weakened. The vicinal diol structures in sugars are capable to complex divalent cations and delocalize them throughout the matrix as long as its glass transition temperature is lower than the phase transition temperature of PM. The loss of divalent cations from bacteriorhodopsin (BR), the only protein in PM, causes the formation of blue membrane (BM), which is accompanied by a loss of beta-sheet structure observable in the infrared spectrum. Glassy sugars are particular useful to observe this transition, as sugar entrapment does not restrict conformational changes of BR but rather retards them. The material obtained was named sugar-induced blue membrane (SIBM). The formation of SIBM is inhibited by the addition of divalent cations. Furthermore, SIBM is reverted immediately to PM by addition of water. A characteristic time dependence of the thermal reversion of SIBM to PM proves that the phase transition of PM triggers the release and uptake of divalent cations and the corresponding color change.

Thermochromism of bacteriorhodopsin and its pH dependence.

Purple membranes (PMs), which consist of the photochromic membrane protein bacteriorhodopsin (BR) and lipids only, show complex thermochromic properties. Three different types of reversible temperature-dependent spectral transitions were found, involving spectral states absorbing at 460, 519, and 630 nm. These thermochromic absorption changes were analyzed in the range from 10 to 80 degrees C. In dependence on the bulk pH value, hypsochromic or bathochromic shifts in the BR absorption spectra are observed in BR gels as well as in BR films. The thermochromic changes between both purple and blue or purple and red were quantified in the CIE color system. The molecular changes causing these effects are discussed, and a model is presented in terms of intramolecular protonation equilibriums. The thermochromic properties of BR may be of interest in applications like security tags, as this feature may complement the well-known photochromic properties of BR.

Purple membranes as microscaled nanopatterned biosubstrates for reversible attachment of biocomponents.

One of the key challenges in the construction of complex bionanotechnological building blocks and devices is the controlled linkage and release of biomolecular components to and from a biomolecular surface. Here we report a versatile, easy and universally applicable method for the reversible assembly of protein-protein conjugates. The process is demonstrated using green fluorescent protein (GFP) and purple membranes (PM) as model compounds. GFP was reversibly bound to PM patches which served as a biomolecular model surface. Due to its size in the micrometre range PM is, as far as its Brownian diffusion is considered, like a solid surface. PMs comprising the mutated bacteriorhodopsin BR-D36C were employed, where aspartic acid in position 36 was replaced with cysteine. The introduced cysteine is accessible from the cytoplasmic side of the membrane. The cysteine group was first functionalized with a nitrilotriacetic acid group (NTA) and then, after loading with Ni, histidine-tagged GFP was bound to the chemically modified PM surface the well-known NTA-His complex. Binding and release of GFP from the PM surface was monitored by atomic force microscopy (AFM).

Two-photon absorption-controlled multidose drug release: a novel approach for secondary cataract treatment.

Tens of millions of cataract surgeries are done every year and the number is increasing heavily. Posterior capsule opacification is the major postoperative complication with an incidence of 10 to 50% within 5 years, depending on the age of the patient. We present a novel approach for secondary cataract treatment in a noninvasive manner. Photochemically triggered drug release from a polymer enables repeated drug applications for cataract treatment years after implantation of the intraocular lens, just when needed. However, light in the visible spectral range must pass through the lens but must not induce drug release. We demonstrate that two-photon absorption photochemistry is a powerful tool to overcome this problem. With wavelengths in the visible regime, a photochemical reaction that requires energies in the UV is triggered. The high intensities needed for this process never occur in any lighting condition in daily lives, but may be easily obtained with focused laser beams routinely used in ophthalmology. The properties of the therapeutic system are specified and the function is demonstrated by in-vitro cell tests. Noninvasive multidose photochemically triggered drug release from implanted intraocular lenses carrying a drug depot may be a therapeutic as well as an economic choice to established treatments of secondary cataracts.

Forming microstructured alkanethiol self-assembled monolayers on gold by laser ablation.

A process to form microstructured alkanethiol self-assembled monolayers (SAMs) on gold is described. It is well known that alkanethiols spontaneously form homogenous SAMs on gold surfaces. By means of laser ablation, the exposed areas of alkanethiol monolayers can be removed from the gold surface. Free gold is obtained which can react further with second and third thiols. By this technique, structured alkanethiol SAMs are obtained reliably and easily. In a rather narrow window of pulse intensities, in our example 120 MW/cm2 +/- 10% from a frequency-doubled Nd: YVO4 laser with 6-ns pulsewidth operating at a repetition rate of 20 kHz, ablation of alkanethiol monolayers is obtained without causing any damage to the gold substrate. Examples are presented where lines down to 10 microm in width were laser ablated into an SAM formed either from a hydrophilic or a hydrophobic alkanethiol and filled with a monolayer of a second alkanethiol of opposite hydrophilicity. The patterned structures were examined by optical and fluorescence microscopy as well as by lateral force microscopy. The presented method enables the preparation of microstructured SAMs on gold and probably on a wide variety of other substrates.

Encapsulation of purple membrane patches into polymeric nanofibers by electrospinning.

The formation of beaded nanofibers containing purple membrane (PM) by electrospinning of polymer solutions is reported. Electrospinning is known as a versatile method to produce polymeric fibers with diameters on the nanometer scale. Embedding of particles significantly larger than the fiber diameter is unexpected because a breakdown of the spinning process is expected when microscaled particles in the polymer solution pass the nozzle. Presumably due to the flexibility of PM patches, the embedding of the membrane patches into the fibers becomes possible. Embedding into nanofibers may be an alternative to microencapsulation for biomolecules.

Biomolecular optical data storage and data encryption.

The use of bacteriorhodopsin (BR) as an active layer in write-once-read-many optical storage is presented. This novel feature of BR materials may be used on a wide variety of substrates, among them transparent substrates but also paper and plastics. The physical basis of the recording process is polarization-sensitive two-photon absorption. As an example for this new BR application, an identification card equipped with an optical recording strip is presented, which has a capacity of about 1 MB of data. The recording density currently used is 125 kB/cm2, which is far from the optical limits but allows operation with cheap terminals using plastic optics. In the examples given, data are stored in blocks of 10 kB each. A special optical encryption procedure allows the stored data to be protected from unauthorized reading. The molecular basis of this property is again the polarization-sensitive recording mechanism. The unique combination of optical storage, photochromism, and traceability of the BR material is combined on the single-molecule level. BR introduces a new quality of storage capability for applications with increased security and anticounterfeiting requirements.

Photochemical preparation of sub-wavelength heterogeneous laser-induced periodic surface structures.

Laser-induced periodic surface structures (LIPSS) are a phenomenon caused by interaction of light with solid surfaces. We present a photochemical concept which uses LIPSS-related light intensity patterns for the generation of heterogeneous nanostructures. The process facilitates arbitrary combinations of substrate and LIPSS-pattern materials. An efficient method for the generation of organometallic hybrid-nanowire arrays on porous anodic aluminum oxide is demonstrated.

Simultaneous removal of thiolated membrane proteins resulting in nanostructured lipid layers.

Self-organization of membrane-embedded peptides and proteins causes the formation of lipid mesostructures in the membranes. One example is purple membranes (PM), which consist of lipids and bacteriorhodopsin (BR) as the only protein component. The BRs form a hexagonal crystalline lattice. A complementary structure is formed by the lipids. Employing BR and PM as an example, we report a method where major parts of the mesoscopic self-assembled protein structures can be extracted from the lipid bilayer membrane. A complementary lipid nanostructure remains on the substrate. To remove such a large number of thiolated proteins simultaneously by applying a mechanical force, they are first reacted at physiological conditions with gold nanoparticles, and then a thin gold film is sputtered onto them that fuses with the gold nanoparticles forming a uniform layer, which finally can be lifted off. In this step, all of the previously gold-labeled proteins are pulled out of the membrane simultaneously. A stable lipid nanostructure is obtained on the mica substrate. Its stability is due to either binding of the lipids to the substrate through ionic bonds or to enough residual proteins to stabilize the lipid nanostructure against reorganization. This method may be applied easily and efficiently wherever thiolated proteins or peptides are employed as self-assembling and structure-inducing units in lipid membranes.

Two-photon absorption of bacteriorhodopsin: formation of a red-shifted thermally stable photoproduct F620.

By means of high-intensity 532 nm laser pulses, a photochemical conversion of the initial B(570) state of bacteriorhodopsin (BR) to a stable photoproduct absorbing maximally at approximately 620 nm in BR suspensions and at approximately 610 nm in BR films is induced. This state, which we named F(620), is photochemically further converted to a group of three products with maximal absorptions in the wavelength range from 340 nm to 380 nm, which show identical spectral properties to the so-called P(360) state reported in the literature. The photoconversion from B(570) to F(620) is most likely a resonant two-photon absorption induced step. The formation of F(620) and P(360) leads to a distinguished photo-induced permanent optical anisotropy in BR films. The spectral dependence of the photo-induced anisotropy and the anisotropy orientations at the educt (B(570)) and product (F(620)) wavelengths are strong indicators that F(620) is formed in a direct photochemical step from B(570). The chemical nature of the P(360) products probably is that of a retro-retinal containing BR, but the structural characteristics of the F(620) state are still unclear. The photo-induced permanent anisotropy induced by short laser pulses in BR films helps to better understand the photochemical pathways related to this transition, and it is interesting in view of potential applications as this feature is the molecular basis for permanent optical data storage using BR films.