Urs Staufer

Microscopy capabilities of the Microscopy, Electrochemistry, and Conductivity Analyzer

The Phoenix microscopy station, designed for the study of Martian dust and soil,consists of a sample delivery system, an optical microscope, and an atomic force microscope. The combination of microscopies facilitates the study of features from the millimeter to nanometer scale. Light-emitting diode illumination allows for full color optical imaging of the samples as well as imaging of ultraviolet-induced visible fluorescence. The atomic force microscope uses an array of silicon tips and can operate in both static and dynamic mode.

Label-Free Detection of Single Protein Molecules and Protein-Protein Interactions Using Synthetic Nanopores

Nanofabricated pores in 20 nm-thick silicon nitride membranes were used to probe various protein analytes as well as to perform an antigen-antibody binding assay. A two-compartment electrochemical cell was separated by a single nanopore, 28 nm in diameter. Adding proteins to one compartment caused current perturbations in the ion current flowing through the pore. These perturbations correlated with both the charge and the size of the protein or of a protein-protein complex. The potential of this nanotechnology for studying protein-protein interactions is highlighted with the sensitive detection of beta-human chorionic gonadotropin, a hormone and clinical biomarker of pregnancy, by monitoring in real time and at a molecular level the formation of a complex between hormones and antibodies in solution. In this form, the assay compared advantageously to immunoassays, with the important difference that labels, immobilization, or amplification steps were no longer needed. In conclusion, we present proof-of-principle that properties of proteins and their interactions can be investigated in solution using synthetic nanopores and that these interactions can be exploited to measure protein concentrations accurately.

Introduction to special section on the Phoenix Mission: Landing Site Characterization Experiments, Mission Overviews, and Expected Science

[1]xa0Phoenix, the first Mars Scout mission, capitalizes on the large NASA investments in the Mars Polar Lander and the Mars Surveyor 2001 missions. On 4 August 2007, Phoenix was launched to Mars from Cape Canaveral, Florida, on a Delta 2 launch vehicle. The heritage derived from the canceled 2001 lander with a science payload inherited from MPL and 2001 instruments gives significant advantages. To manage, build, and test the spacecraft and its instruments, a partnership has been forged between the Jet Propulsion Laboratory, the University of Arizona (home institution of principal investigator P. H. Smith), and Lockheed Martin in Denver; instrument and scientific contributions from Canada and Europe have augmented the mission. The science mission focuses on providing the ground truth for the 2002 Odyssey discovery of massive ice deposits hidden under surface soils in the circumpolar regions. The science objectives, the instrument suite, and the measurements needed to meet the objectives are briefly described here with reference made to more complete instrument papers included in this special section. The choice of a landing site in the vicinity of 68°N and 233°E balances scientific value and landing safety. Phoenix will land on 25 May 2008 during a complex entry, descent, and landing sequence using pulsed thrusters as the final braking strategy. After a safe landing, twin fan-like solar panels are unfurled and provide the energy needed for the mission. Throughout the 90-sol primary mission, activities are planned on a tactical basis by the science team; their requests are passed to an uplink team of sequencing engineers for translation to spacecraft commands. Commands are transmitted each Martian morning through the Deep Space Network by way of a Mars orbiter to the spacecraft. Data are returned at the end of the Martian day by the same path. Satisfying the missions goals requires digging and providing samples of interesting layers to three on-deck instruments. By verifying that massive water ice is found near the surface and determining the history of the icy soil by studying the mineralogical, chemical, and microscopic properties of the soil grains, Phoenix will address questions concerning the effects of climate change in the northern plains. A conclusion that unfrozen water has modified the soil naturally leads to speculation as to the biological potential of the soil, another scientific objective of the mission.

Assessment of insulated conductive cantilevers for biology and electrochemistry

This paper describes the characterization and application of electrically insulated conductive tips mounted on a cantilever for use in an atomic force microscope and operated in liquid. These multifunctional probes were microfabricated and designed for measurements on biological samples in buffer solution, but they can also be employed for electrochemical applications, in particular scanning electrochemical microscopy. The silicon nitride based cantilevers had a spring constant ≤0.1 N m-1 and a conductive tip, which was insulated except at the apex. The conductive core of the tip consisted of a metal, e.g. platinum silicide, and exhibited a typical radius of 15 nm. The mechanical and electrical characterization of the probe is presented and discussed. First measurements on the hexagonally packed intermediate layer of Deinococcus radiodurans demonstrated the possibility to adjust the image contrast by applying a voltage between a support and the conductive tip and to measure variations of less than 1 pA in faradaic current with a lateral resolution of 7.8 nm.

Conductive supports for combined AFM–SECM on biological membranes

Four different conductive supports are analysed regarding their suitability for combined atomic force and scanning electrochemical microscopy (AFM-SECM) on biological membranes. Highly oriented pyrolytic graphite (HOPG), MoS(2), template stripped gold, and template stripped platinum are compared as supports for high resolution imaging of reconstituted membrane proteins or native membranes, and as electrodes for transferring electrons from or to a redox molecule. We demonstrate that high resolution topographs of the bacterial outer membrane protein F can be recorded by contact mode AFM on all four supports. Electrochemical feedback experiments with conductive cantilevers that feature nanometre-scale electrodes showed fast re-oxidation of the redox couple Ru(NH(3))(6)(3+/2+) with the two metal supports after prolonged immersion in electrolyte. In contrast, the re-oxidation rates decayed quickly to unpractical levels with HOPG or MoS(2) under physiological conditions. On HOPG we observed heterogeneity in the re-oxidation rate of the redox molecules with higher feedback currents at step edges. The latter results demonstrate the capability of conductive cantilevers with small electrodes to measure minor variations in an SECM signal and to relate them to nanometre-scale features in a simultaneously recorded AFM topography. Rapid decay of re-oxidation rate and surface heterogeneity make HOPG or MoS(2) less attractive for combined AFM-SECM experiments on biological membranes than template stripped gold or platinumxa0supports.

Controlled, Reversible, and Nondestructive Generation of Uniaxial Extreme Strains (>10%) in Graphene

Theoretical calculations have predicted that extreme strains (>10%) in graphene would result in novel applications. However, up to now the highest reported strain reached ∼1.3%. Here, we demonstrate uniaxial strains >10% by pulling graphene using a tensile-MEMS. To prevent it from slipping away it was locally clamped with epoxy using a femtopipette. The results were analyzed using Raman spectroscopy and optical tracking. Furthermore, analysis proved the process to be reversible and nondestructive for the graphene.

Effects of size and defects on the elasticity of silicon nanocantilevers

The size-dependent elastic behavior of silicon nanocantilevers and nanowires, specifically the effective Youngs modulus, has been determined by experimental measurements and theoretical investigations. The size dependence becomes more significant as the devices scale down from micro- to nano-dimensions, which has mainly been attributed to surface effects. However, discrepancies between experimental measurements and computational investigations show that there could be other influences besides surface effects. In this paper, we try to determine to what extent the surface effects, such as surface stress, surface elasticity, surface contamination and native oxide layers, influence the effective Youngs modulus of silicon nanocantilevers. For this purpose, silicon cantilevers were fabricated in the top device layer of silicon on insulator (SOI) wafers, which were thinned down to 14 nm. The effective Youngs modulus was extracted with the electrostatic pull-in instability method, recently developed by the authors (H Sadeghian et al 2009 Appl. Phys. Lett. 94 221903). In this work, the drop in the effective Youngs modulus was measured to be significant at around 150 nm thick cantilevers. The comparison between theoretical models and experimental measurements demonstrates that, although the surface effects influence the effective Youngs modulus of silicon to some extent, they alone are insufficient to explain why the effective Youngs modulus decreases prematurely. It was observed that the fabrication-induced defects abruptly increased when the device layer was thinned to below 100 nm. These defects became visible as pinholes during HF-etching. It is speculated that they could be the origin of the reduced effective Youngs modulus experimentally observed in ultra-thin silicon cantilevers.

Design and fabrication of nanofluidic devices by surface micromachining

Using surface micromachining technology, we fabricated nanofluidic devices with channels down to 10xa0nm deep, 200xa0nm wide and up to 8xa0cm long. We demonstrated that different materials, such as silicon nitride, polysilicon and silicon dioxide, combined with variations of the fabrication procedure, could be used to make channels both on silicon and glass substrates. Critical channel design parameters were also examined. With the channels as the basis, we integrated equivalent elements which are found on micro total analysis (μTAS) chips for electrokinetic separations. On-chip platinum electrodes enabled electrokinetic liquid actuation. Micro-moulded polydimethylsiloxane (PDMS) structures bonded to the devices served as liquid reservoirs for buffers and sample. Ionic conductance measurements showed Ohmic behaviour at ion concentrations above 10xa0mM, and surface charge governed ion transport below 5xa0mM. Low device to device conductance variation (1%) indicated excellent channel uniformity on the wafer level. As proof of concept, we demonstrated electrokinetic injections using an injection cross with volume below 50xa0attolitres (10(-18)xa0l).

Selective molecular assembly patterning at the nanoscale: a novel platform for producing protein patterns by electron-beam lithography on Sio2/indium tin oxide coated glass substrates

The creation of geometrically well-defined submicron structures on insulating substrates by e-beam lithography is hampered by surface charging. This problem becomes crucial when trying to create nanosized protein patterns by selective molecular assembly patterning (SMAP) on transparent glass substrates. In this paper we demonstrate that the use of thin films of conductive indium tin oxide resolves the issue of surface charging during e-beam writing while being compatible with the standard SMAP protocol for surface modification.

Sub-ppm detection of vapors using piezoresistive microcantilever array sensors

The performance of microfabricated piezoresistive cantilever array sensors has been evaluated using various vapors of volatile organic compounds including alkanes with different chain length from 5 (n-pentane) to 14 (n-tetradecane). We demonstrate that piezoresistive microcantilever array sensors have the selectivity of discriminating individual alkanes in a homologous series as well as common volatile organic compounds according to principal component analysis. We developed a new method to evaluate the sensitivity, taking advantage of the low vapor pressures of alkanes with longer chains, such as n-dodecane, n-tridecane and n-tetradecane, under saturated vapor conditions. This method reveals sub-ppm sensitivity and the cantilever response is found to follow the mass of evaporated analytes as calculated using a quantitative model based on the Langmuir evaporation model.