Bohuslav Rezek

Diamond for bio-sensor applications

A summary of photo- and electrochemical surface modifications applied on single-crystalline chemical vapour deposition (CVD) diamond films is given. The covalently bonded formation of amine- and phenyl-linker molecule layers is characterized using x-ray photoelectron spectroscopy, atomic force microscopy (AFM), cyclic voltammetry and field-effect transistor characterization experiments. Amine- and phenyl-layers are very different with respect to formation, growth, thickness and molecule arrangement. We detect a single-molecular layer of amine-linker molecules on diamond with a density of about 1014?cm?2 (10% of carbon bonds). Amine molecules are bonded only on initially H-terminated surface areas to carbon. In the case of electrochemical deposition of phenyl-layers, multi-layer formation is detected due to three-dimensional (3D) growths. This gives rise to the formation of typically 25?? thick layers. The electrochemical grafting of boron-doped diamond works on H-terminated and oxidized surfaces.After reacting such films with hetero-bifunctional crosslinker molecules, thiol-modified ss-DNA markers are bonded to the organic system. Application of fluorescence and AFM on hybridized DNA films shows dense arrangements with densities of up to 1013?cm?2. The DNA is tilted by an angle of about 35? with respect to the diamond surface. Shortening the bonding time of thiol-modified ss-DNA to 10?min causes a decrease of DNA density to about 1012?cm?2. Application of AFM scratching experiments shows threshold removal forces of around 75?nN for DNA bonded on phenyl-linker molecules and of about 45?nN for DNA bonded to amine-linker molecules. DNA sensor applications using Fe(CN6)3?/4? mediator redox molecules, impedance spectroscopy and DNA-field effect transistor devices performances are introduced and discussed.

Diamond and biology

A summary of photo- and electrochemical surface modifications applied on single-crystalline chemical vapour deposition diamond films is given. The covalently bonded formation of amine and phenyl linker molecular layers is characterized using X-ray photoelectron spectroscopy, atomic force microscopy (AFM), cyclic voltammetry and field-effect transistor characterization experiments. Amine and phenyl layers are very different with respect to formation, growth, thickness and molecular arrangement. We deduce a sub-monolayer of amine linker molecules on diamond with approximately 10% coverage of 1.5×1015 cm−2 carbon bonds. Amine is bonded only on initially H-terminated surface areas. In the case of electrochemical deposition of phenyl layers, multilayer properties are detected with three-dimensional nitrophenyl growth properties. This leads to the formation of typically 25 Å thick layers. The electrochemical bonding to boron-doped diamond works on H-terminated and oxidized surfaces. After reacting such films with heterobifunctional cross-linker molecules, thiol-modified ss-DNA markers are bonded to the organic system. Application of fluorescence and AFM on hybridized DNA films shows dense arrangements with densities up to 1013 cm−2. The DNA is tilted by an angle of approximately 35° with respect to the diamond surface. Shortening the bonding time of thiol-modified ss-DNA to 10 min causes a decrease in DNA density to approximately 1012 cm−2. Application of AFM scratching experiments shows threshold removal forces of approximately 75 and 45 nN for the DNA bonded to the phenyl and the amine linker molecules, respectively. First, DNA sensor applications using Fe(CN6)3−/4− mediator redox molecules and DNA field-effect transistor devices are introduced and discussed.

Fermi level on hydrogen terminated diamond surfaces

Atomic force microscopy and Kelvin probe experiments are applied to characterize hydrogen terminated patterns contacted with gold and aluminum on (100) diamond surfaces. On hydrogen terminated diamond the work function of 4.9 eV is detected, with an accuracy of about 0.1 eV. Taking into account the negative electron affinity of −1.3 eV and a band gap of 5.5 eV the Fermi energy is 0.7 eV deep in the valence band. Illumination of the sample results in a shift of the surface Fermi level by as much as 0.2 eV. This is attributed to a surface photovoltage effect.

Nanoscale topography of nanocrystalline diamonds promotes differentiation of osteoblasts.

The excellent mechanical, tribological and biochemical properties of diamond coatings are promising for improving orthopedic or stomatology implants. A crucial prerequisite for such applications is an understanding and control of the biological response of the diamond coatings. This study concentrates on the correlation of diamond surface properties with osteoblast behavior. Nanocrystalline diamond (NCD) films (grain size up to 200 nm, surface roughness 20 nm) were deposited on silicon substrates of varying roughnesses (1, 270 and 500 nm) and treated by oxygen plasma to generate a hydrophilic surface. Atomic force microscopy was used for topographical characterization of the films. As a reference surface, tissue culture polystyrene (PS) was used. Scanning electron microscopy and immunofluorescence staining was used to visualize cell morphological features as a function of culture time. Metabolic activity, alkaline phosphatase activity, and calcium and phosphate deposition was also monitored. The results show an enhanced osteoblast adhesion as well as increased differentiation (raised alkaline phosphatase activity and mineral deposition) on NCD surfaces (most significantly on RMS 20 nm) compared to PS. This is attributed mainly to the specific surface topography as well as to the biocompatible properties of diamond. Hence the controlled (topographically structured) diamond coating of various substrates is promising for preparation of better implants, which offer faster colonization by specific cells as well as longer-term stability.

Local characterization of electronic transport in microcrystalline silicon thin films with submicron resolution

Two-dimensional maps of dark conductivity with submicron resolution have been obtained on in situ prepared hydrogenated microcrystalline silicon (μc-Si:H) layers used for solar cells by atomic force microscopy with conductive cantilever. Comparison of the morphology and current image allows clear identification of Si crystallites. Pronounced current decrease has been detected at the grain boundaries. The technique was used to study initial stages of μc-Si:H growth, and we show how the incubation layer, detrimental for solar cells efficiency, can be minimized by pulsed excimer laser crystallization of the initial amorphous layer.

Micro-Pattern Guided Adhesion of Osteoblasts on Diamond Surfaces

Microscopic chemical patterning of diamond surfaces by hydrogen and oxygen surface atoms is used for self-assembly of human osteoblastic cells into micro-arrays. The cell adhesion and assembly is further controlled by concentration of cells (2,500-10,000 cells/cm2) and fetal bovine serum (0-15%). The cells are characterized by fluorescence microscopy of actin fibers and nuclei. The serum protein adsorption is studied by atomic force microscopy (AFM). The cells are arranged selectively on O-terminated patterns into 30-200 μm wide arrays. Higher cell concentrations allow colonization of unfavorable H-terminated regions due to mutual cell communication. There is no cell selectivity without the proteins in the medium. Based on the AFM, the proteins are present on both H- and O-terminated surfaces. Pronounced differences in their thickness, surface roughness, morphology, and phase images indicate different conformation of the proteins and explain the cell selectivity.

Basic features of transport in microcrystalline silicon

Charge transport in microcrystalline silicon is strongly influenced by its heterogeneous microstructure composed of crystalline grains and amorphous tissue. An even bigger effect on transport is their arrangement in grain aggregates or possibly columns, separated by grain boundaries, causing transport anisotropy and/or depth profile of transport properties. We review special experimental methods developed to study the resulting transport features: local electronic studies by combined atomic force microscopy, anisotropy of conductivity and diffusion length and also their thickness dependence. A simple model based on the concept of changes of transport path for description of the observed phenomena is reviewed and its consequences for charge collection in microcrystalline based solar cells are discussed.

Microcrystalline silicon thin films studied by atomic force microscopy with electrical current detection

Hydrogenated microcrystalline silicon (μc-Si:H) layers with thickness from 100 to 540 nm were prepared in situ by plasma enhanced chemical vapor deposition. The growth of μc-Si:H on various substrates [NiCr, device quality, and laser annealed amorphous silicon (a-Si:H)] was studied in ultrahigh vacuum by atomic force microscope using a conductive cantilever which enabled simultaneous measurement of morphology and local current with lateral resolution below 5 nm. The effect of barriers, voltage, and time on contrast in local current map is discussed in detail. Coexistent amorphous and microcrystalline regions are clearly identified due to their different conductivity. Laser annealing of the a-Si:H substrate significantly increases the crystalline fraction at the same layer thickness. Grains as small as 10–30 nm separated by less conductive grain boundaries were revealed in microcrystalline regions.

Enhanced Growth and Osteogenic Differentiation of Human Osteoblast-Like Cells on Boron-Doped Nanocrystalline Diamond Thin Films

Intrinsic nanocrystalline diamond (NCD) films have been proven to be promising substrates for the adhesion, growth and osteogenic differentiation of bone-derived cells. To understand the role of various degrees of doping (semiconducting to metallic-like), the NCD films were deposited on silicon substrates by a microwave plasma-enhanced CVD process and their boron doping was achieved by adding trimethylboron to the CH4:H2 gas mixture, the B∶C ratio was 133, 1000 and 6700 ppm. The room temperature electrical resistivity of the films decreased from >10 MΩ (undoped films) to 55 kΩ, 0.6 kΩ, and 0.3 kΩ (doped films with 133, 1000 and 6700 ppm of B, respectively). The increase in the number of human osteoblast-like MG 63 cells in 7-day-old cultures on NCD films was most apparent on the NCD films doped with 133 and 1000 ppm of B (153,000±14,000 and 152,000±10,000 cells/cm2, respectively, compared to 113,000±10,000 cells/cm2 on undoped NCD films). As measured by ELISA per mg of total protein, the cells on NCD with 133 and 1000 ppm of B also contained the highest concentrations of collagen I and alkaline phosphatase, respectively. On the NCD films with 6700 ppm of B, the cells contained the highest concentration of focal adhesion protein vinculin, and the highest amount of collagen I was adsorbed. The concentration of osteocalcin also increased with increasing level of B doping. The cell viability on all tested NCD films was almost 100%. Measurements of the concentration of ICAM-1, i.e. an immunoglobuline adhesion molecule binding inflammatory cells, suggested that the cells on the NCD films did not undergo significant immune activation. Thus, the potential of NCD films for bone tissue regeneration can be further enhanced and tailored by B doping and that B doping up to metallic-like levels is not detrimental for cells.

Size and Purity Control of HPHT Nanodiamonds down to 1 nm

High-pressure high-temperature (HPHT) nanodiamonds originate from grinding of diamond microcrystals obtained by HPHT synthesis. Here we report on a simple two-step approach to obtain as small as 1.1 nm HPHT nanodiamonds of excellent purity and crystallinity, which are among the smallest artificially prepared nanodiamonds ever shown and characterized. Moreover we provide experimental evidence of diamond stability down to 1 nm. Controlled annealing at 450 °C in air leads to efficient purification from the nondiamond carbon (shells and dots), as evidenced by X-ray photoelectron spectroscopy, Raman spectroscopy, photoluminescence spectroscopy, and scanning transmission electron microscopy. Annealing at 500 °C promotes, besides of purification, also size reduction of nanodiamonds down to ∼1 nm. Comparably short (1 h) centrifugation of the nanodiamonds aqueous colloidal solution ensures separation of the sub-10 nm fraction. Calculations show that an asymmetry of Raman diamond peak of sub-10 nm HPHT nanodiamonds can be well explained by modified phonon confinement model when the actual particle size distribution is taken into account. In contrast, larger Raman peak asymmetry commonly observed in Raman spectra of detonation nanodiamonds is mainly attributed to defects rather than to the phonon confinement. Thus, the obtained characteristics reflect high material quality including nanoscale effects in sub-10 nm HPHT nanodiamonds prepared by the presented method.