Nonlinear optomechanical measurement of mechanical motion.

Precision measurement of nonlinear observables is an important goal in all facets of quantum optics. This allows measurement-based non-classical state preparation, which has been applied to great success in various physical systems, and provides a route for quantum information processing with otherwise linear interactions. In cavity optomechanics much progress has been made using linear interactions and measurement, but observation of nonlinear mechanical degrees-of-freedom remains outstanding. Here we report the observation of displacement-squared thermal motion of a micro-mechanical resonator by exploiting the intrinsic nonlinearity of the radiation-pressure interaction. Using this measurement we generate bimodal mechanical states of motion with separations and feature sizes well below 100 pm. Future improvements to this approach will allow the preparation of quantum superposition states, which can be used to experimentally explore collapse models of the wavefunction and the potential for mechanical-resonator-based quantum information and metrology applications.

Micromechanical PDGF recognition via lab-on-a-disc aptasensor arrays.

A plug-and-play CD-like platform is used to perform a statistical detection of platelet derived growth factor (PDGF) proteins through aptamer-based surface functionalization of multiple microcantilever arrays. When PDGF proteins bind to aptamer coatings, the cantilevers deflect. The deflection response is monitored by optical read-out units from a commercial DVD-ROM device. We report on the use of an improved sensing platform which facilitates measurements under continuous liquid flow and with temperature control. Also, the mechanical wobbling of the DVD-ROM platform has been minimized and the scanning system has been optimized in order to detect cantilever deflections in liquid with nanometer scale resolution. The capability of the sensing platform is demonstrated by detection of clinically relevant concentrations of PDGF proteins. We present statistical measurements on 100 microcantilevers at different concentrations of PDGF, ranging from 10 nM to 400 nM. Hereby it is possible to reliably characterize the averaged mechanical response of cantilevers as a function of protein concentration.

The influence of refractive index change and initial bending of cantilevers on the optical lever readout method.

It has been speculated that the initial bending of cantilevers has a major influence on the detector signal in a cantilever-based sensor using the optical lever readout method. We have investigated theoretically as well as experimentally the changes induced in the detector signal when the optical lever technique is used to monitor a cantilever with initial bending during changes in the refractive index of the surrounding media. We find that for changes in refractive index as small as 10(-4) the detector signal is highly dependent on the initial bending of the cantilever. The findings are validated experimentally using an environmental chamber and varying the pressure. We sketch routes to circumvent the problem and formulas suitable for data treatment are given.

Mass and position determination of attached particles on cantilever based mass sensors.

An analytical expression relating mass and position of a particle attached on a cantilever to the resulting change in cantilever resonant frequency is derived. Theoretically, the position and mass of the attached particle can be deduced by combining measured resonant frequencies of several bending modes. This finding is verified experimentally using a microscale cantilever with and without an attached gold bead. The resonant frequencies of several bending modes are measured as a function of the bead position. The bead mass and position calculated from the measured resonant frequencies are in good agreement with the expected mass and the position measured.

Highly sensitive polymer-based cantilever-sensors for DNA detection.

We present a technology for the fabrication of cantilever arrays aimed to develop an integrated biosensor microsystem. The fabrication process is based on spin coating of the photosensitive polymer and near-ultraviolet exposure. Arrays of up to 33 microcantilevers are fabricated in the novel polymer material SU-8. The low Young's modulus of the polymer, 40 times lower than that of silicon, enables to improve the sensitivity of the sensor device for target detection. The mechanical properties of SU-8 cantilevers, such as spring constant, resonant frequency and quality factor are characterized as a function of the dimensions and the medium. The devices have been tested for measurement of the adsorption of single stranded DNA and subsequent interstitial adsorption of lateral spacer molecules. We demonstrate that sensitivity is enhanced by a factor of six compared to that of commercial silicon nitride cantilevers.

On the electromechanical modelling of a resonating nano-cantilever-based transducer.

An electromechanical model for a transducer based on a lateral resonating cantilever is described. The on-plane vibrations of the cantilever are excited electrostatically by applying DC and AC voltages from a driver electrode placed closely parallel to the cantilever. The model predicts the static deflection and the frequency response of the oscillation amplitude for different voltage polarization conditions. For the electrostatic force calculation the model takes into account the real deflection shape of the cantilever and the contribution to the cantilever-driver capacitance of the fringing field. Both the static and dynamic predictions have been validated experimentally by measuring the deflection of the cantilever by means of an optical microscope.

Atomic force microscope characterization of a resonating nanocantilever.

An atomic force microscope (AFM) is used as a nanometer-scale resolution tool for the characterization of the electromechanical behaviour of a resonant cantilever-based mass sensor. The cantilever is actuated electrostatically by applying DC and AC voltages from a driver electrode placed closely parallel to the cantilever. In order to minimize the interaction between AFM probe and the resonating transducer cantilever, the AFM is operated in a dynamic non-contact mode, using oscillation amplitudes corresponding to a low force regime. The dependence of the static cantilever deflection on DC voltage and of the oscillation amplitude on the frequency of the AC voltage is measured by this technique and the results are fitted by a simple non-linear electromechanical model.

Hybridisation of short DNA molecules investigated with in situ atomic force microscopy.

By introducing the complementary DNA (cDNA) strand to a molecular layer of short single stranded DNA (ssDNA), immobilised on a gold surface, we have investigated hybridisation between the two DNA strands through the technique of in situ atomic force microscopy (AFM). Before introduction of cDNA, the ssDNA molecular layer was modulated with the spacer molecule mercaptohexanol (MCH), which makes the ssDNA molecules more accessible for hybridisation. With in situ AFM, we have monitored the formation of a smooth, mixed molecular layer containing ssDNA and MCH. Furthermore, the hybridisation between the two DNA strands has been studied. Introduction of the cDNA strand resulted in an increase in smoothness and thickness of the molecular layer. Both the increase in order and thickness of the molecular layer can be expected if hybridisation occurs, since double stranded DNA molecules have a more rigid and elongated structure than ssDNA molecules.

Optimised cantilever biosensor with piezoresistive read-out.

We present a cantilever-based biochemical sensor with piezoresistive read-out which has been optimised for measuring surface stress. The resistors and the electrical wiring on the chip are encapsulated in low-pressure chemical vapor deposition (LPCVD) silicon nitride, so that the chip is well suited for operation in liquids. The wiring is titanium silicide which-in contrast to conventional metal wiring-is compatible with the high-temperature LPCVD coating process.

AFM lithography of aluminum for fabrication of nanomechanical systems.

Nanolithography by local anodic oxidation of surfaces using atomic force microscopy (AFM) has proven to be more reproducible when using dynamic, non-contact mode. Hereby, the tip/sample interaction forces are reduced dramatically compared to contact mode, and thus tip wear is greatly reduced. Anodic oxidation of Al can be used for fabricating nanomechanical systems, by using the Al oxide as a highly selective dry etching mask. In our experiments, areas as large as 2 micro m x 3 micro m have been oxidized repeatedly without any sign of tip-wear. Furthermore, line widths down to 10nm have been routinely obtained, by optimization of AFM parameters, such as tip/sample distance, voltage and scan speed. Finally, AFM oxidation experiments have been performed on CMOS processed chips, demonstrating the first steps of fabricating fully functional nanomechanical devices.

Sodium and chloride transport in soft water and hard water acclimated zebrafish (Danio rerio).

While the zebrafish is commonly used for studies of developmental biology and toxicology, very little is known about their osmoregulatory physiology. The present investigation of Na(+) and Cl(-) transport revealed that the zebrafish is able to tolerate extremely low ambient ion concentrations and that this is achieved at least in part by a greatly enhanced apparent uptake capacity and affinity for both ions. Zebrafish maintain plasma and whole body electrolyte concentrations similar to most other freshwater teleosts even in deionized water containing only 35 microM NaCl, i.e soft water. We recorded an extremely low transport affinity constant (K(m)) of 8+/-1 microM for the active uptake of Cl(-) in soft water acclimated fish, while other transport kinetic parameters were in agreement with reports for other freshwater organisms. While both Na(+) and Cl(-) uptake in soft water clearly depends on apical proton pump activity, changes in abundance and possibly localization of this protein did not appear to contribute to soft water acclimation. Active Cl(-) uptake was strongly dependent on branchial carbonic anhydrase (CA) activity regardless of water type, while the response of Na(+) transport to a CA inhibitor was more variable. Differential response of Na(+) uptake to amiloride depending on acclimation medium suggests that different Na(+) transport mechanisms are employed by zebrafish acclimated to soft and hard water.

Structural characterisation of the H-bonded [MnII(OH2)2...(OCH3)2MnIII] motif: a model for resting state hydroxylic solvent coordination in M(II)-M(III) enzymes.

A mixed-valence dimanganese complex with two water ligands bound to the Mn(II) ion and two methoxide ligands bound to the Mn(III) ion in a H-bonded arrangement has been structurally characterised and models possible resting state water/hydroxide coordination in purple acid phosphatases.

Environmental sensors based on micromachined cantilevers with integrated read-out

An AFM probe with integrated piezoresistive read-out has been developed and applied as a cantilever-based environmental sensor. The probe has a built-in reference cantilever, which makes it possible to subtract background drift directly in the measurement. Moreover, the integrated read-out facilitates measurements in liquid. The probe has been successfully implemented in gaseous as well as in liquid experiments. For example, the probe has been used as an accurate and minute thermal sensor and as a humidity sensor. In liquid, the probe has been used to detect the presence of alcohol in water.

Primary structure of EPV20, a secretory glycoprotein containing a previously uncharacterized type of domain.

A 20-kDa glycoprotein, EPV20, was isolated from bovine milk and characterized. The primary structure was determined by cDNA and protein sequencing combined with mass spectrometry. EPV20 is a 130-residue polypeptide synthesized with a 19-residue signal peptide. The function of EPV20 is unknown, but it displays 79% sequence similarity to a putative protein deduced from a human testis cDNA sequence designated HE1 (human epididymis clone 1) (Kirchhoff, C., 1992. EMBL/GeneBank/DDBJ Databases, accession number X67698). Northern blot analysis showed the bovine EPV20 to be expressed in kidney, spleen, liver and mammary gland, but remarkably not in bovine testis. The six Cys residues of EPV20 were found to be disulfide-linked in a 1-6, 2-3 and 4 5 pattern. This disulfide arrangement has been observed in other proteins, e.g. in human prostatic acid phosphatase, but the spacing between the cystines differs. Therefore, EPV20 represents a new structure among the large group of proteins containing domains with three disulfide bonds.

Procathepsin D cannot autoactivate to cathepsin D at acid pH.

The amino acid sequence of the propart of bovine procathepsin D was determined at the protein level. Incubation of the isolated procathepsin D at pH 3.5-5.0 for 30-120 min leads to a 2 kDa reduction in its molecular mass, as seen by SDS-PAGE. The activation product is pseudocathepsin D and is the result of a proteolytic cleavage between LeuP26 and IleP27 in the propart. Incubation at pH 5.0 for 20 h of either procathepsin D or pseudocathepsin D results in both cases in approximately equal amounts of pseudocathepsin D and a further processed intermediate, nine amino acids shorter than pseudocathepsin D. No reaction products corresponding to cathepsin D with a mature amino terminus were observed, showing that autoproteolysis alone cannot generate the mature form found in the lysosomes.