J. Heitz

Proliferation of aligned mammalian cells on laser-nanostructured polystyrene.

Biomaterial surface chemistry and nanoscale topography are important for many potential applications in medicine and biotechnology as they strongly influence cell function, adhesion and proliferation. In this work, we present periodic surface structures generated by linearly polarized KrF laser light (248 nm) on polystyrene (PS) foils. These structures have a periodicity of 200-430 nm and a depth of 30-100 nm, depending on the angle of incidence of the laser beam. The changes in surface topography and chemistry were analysed by atomic force microscopy (AFM), advancing water contact-angle measurements, Fourier-transform infrared spectroscopy using an attenuated total reflection device (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). We show that the surface laser modification results in a significantly enhanced adhesion and proliferation of human embryonic kidney cells (HEK-293) compared to the unmodified polymer foil. Furthermore, we report on the alignment of HEK-293 cells, Chinese hamster ovary (CHO-K1) cells and skeletal myoblasts along the direction of the structures. The results indicate that the presence of nanostructures on the substrates can guide cell alignment along definite directions, and more importantly, in our opinion, that this alignment is only observed when the periodicity is above a critical periodicity value that is cell-type specific.

Rapid in-situ analysis of liquid steel by laser-induced breakdown spectroscopy☆

Abstract Laser-induced breakdown spectroscopy (LIBS) denotes a technique where a pulsed laser beam is used to ablate small amounts of the target material. The characteristic optical emission line intensities of the excited species in the laser-generated plasma allow a quantitative chemical analysis of the target material. LIBS is a fast, non-contact method allowing large working distances between the sample under investigation and the detection system. These properties make LIBS applicable to process control in metallurgy. We describe an apparatus designed for rapid in-situ analysis of solid and molten metals at variable distances of up to 1.5 m. A variable lens system allows compensation for varying positions of the liquid steel surface. The LIBS signal is guided by a fiber optic bundle of 12-m length to the spectrometer. Analysis of an elements concentration takes 7 s. Laboratory experiments using an induction furnace showed that the addition of admixtures to liquid steel results in rapid response of the system. Results including the in-situ monitoring of Cr, Cu, Mn and Ni within certain concentration ranges are presented (Cr: 0.11–13.8 wt.%; Cu: 0.044–0.54 wt.%; Mn: 1.38–2.5 wt.%; Ni: 0.049–5.92 wt.%).

Adhesion and proliferation of human endothelial cells on photochemically modified polytetrafluoroethylene

We studied the adhesion and proliferation of human endothelial cells on photochemically modified polytetrafluoroethylene samples. The polymer surfaces were modified by exposure to the ultraviolet light of a Xe(2)(*)-excimer lamp at a wavelength of 172 nm in an ammonia atmosphere. Treatment times were between 10 and 20 min. The endothelial cell density was determined 1, 3 and 8 days after seeding by image analysis. Surface modification of the samples resulted in a significant increase in the number of adhering cells and in the formation of a confluent cell layer after 3-8 days. The results were comparable than those obtained on polystyrene Petri dishes, which are used as standard substrates in cell cultivation. Thus modified PTFE appears to be a promising material for the fabrication of artificial vascular prostheses coated with endothelial cells.

Pulsed-laser deposition of crystalline Teflon (PTFE) films

Abstract Thin films of crystalline polytetrafluoroethylene (PTFE) were prepared by pulsed-laser deposition using 248 nm UV-excimer-laser radiation. Pressed powder pellets and bulk PTFE have been employed as target material. The films were analyzed by means of optical polarization microscopy, stylus profilometry, capacity measurements, XRD, and IR spectroscopy. The effect of substrate temperature T s on the morphology and crystallinity of the films was studied. Films deposited from pressed powder targets at sufficiently high T s consist mainly of spherulite-like microcrystallites. These films are continuous, pinhole-free, well adherent to the substrate, and have a composition which is similar to that of the target material. It is suggested that film formation is based on laser-assisted material transfer with subsequent melting and crystallization. They are superior to films deposited from PTFE bulk targets, cut from a solid rod, with respect to film morphology, deposition rate, film cohesion, and optical and electrical properties.

Simple and versatile methods for the fabrication of arrays of live mammalian cells

Single-step methods for the generation of patterned surfaces on hydrogels are presented. Poly(vinyl alcohol) films covalently bonded on glass cover slips and commercially available hydrogel-coated polystyrene plates were used as cell-repellent surfaces. Cell-adhesive domains were created by spotting dilute solutions of sodium hypochlorite onto the surfaces. Alternatively, domains supporting cell attachment were created by exposure to UV light from a xenon excimer lamp, employing a contact mask. Rat skeletal myoblast cells, HEK 293 human embryonic kidney cells and Caco-2 colon carcinoma cells adhered and spread exclusively on modified areas. The surfaces are durable for weeks under cell culture conditions and re-usable after removal of the cells by trypsin treatment. Arrays of adhesive spots seeded with cells at a low density permitted dynamic monitoring of cell proliferation. Selected colonies can be harvested from the surfaces by means of local trypsination. Thus, these techniques may provide useful tools for the isolation of clonal cell populations. Additionally, we demonstrate the possibility of surface-mediated gene delivery from the micro patterns. We show that DNA, complexed with a lipid reagent, can be adsorbed on modified poly(vinyl alcohol) coatings, resulting in spatially controlled adhesion and reverse transfection of HEK 293 cells.

Structure formation in UV-laser ablated poly-ethylene-terephthalate (PET)

Structure formation observed in UV-laser ablated poly-ethylene-terephthalate (PET) foils can uniquely be assigned to mechanical and thermal pretreatments.

Charge stability of pulsed-laser deposited polytetrafluoroethylene film electrets

Pulsed-laser deposited (PLD) polytetrafluoroethylene (Teflon-PTFE) films from press-sintered powder targets are found to be highly crystalline, with spherulite sizes adjustable over more than one order of magnitude by suitable thermal annealing. Films with large spherulites show an excellent charge stability, comparable and even superior to commercially available Teflon-PTFE foils. PLD-PTFE enlarges the family of Teflon materials and may thus become interesting for potential miniaturized electret devices.

Near-field optical nanopatterning of crystalline silicon

Nanoscale photochemical and photophysical etching of Si in Cl2 atmosphere is demonstrated by means of an optical near-field setup. With 351 nm Ar+-laser radiation and low intensities, the etching mechanism is purely photochemical. In this regime, the width of patterns—which is about 115 nm at full width at half maximum (FWHM)—corresponds, approximately, to the diameter of the fiber tip. The vertical etch rate is of the order of 1 nm/s. With 514.5 nm Ar+-laser light etching is observed only at significantly higher laser-light intensities. Patterns with width down to about 30 nm at FWHM have been achieved. Here, the lateral resolution corresponds to about 1/18 of the laser wavelength employed.

Enhanced adhesion of metal films on PET after UV-laser treatment

The adhesion-force of thin metal films on PET foils can be significantly improved by UV excimer-laser irradiation of the polymer surface prior to metal deposition. The laser fluences required are well below the ablation threshold.

Femtosecond excimer-laser-induced structure formation on polymers

Structure formation upon 500 fs 248 nm KrF-laser irradiation of PolyEthylene Terephthalate (PET) and PolyImide (PI) has been investigated. The results obtained with fs pulses have been compared to those with ns pulses.