Philippe Niedermann

FluidFM: Combining Atomic Force Microscopy and Nanofluidics in a Universal Liquid Delivery System for Single Cell Applications and Beyond

We describe the fluidFM, an atomic force microscope (AFM) based on hollow cantilevers for local liquid dispensing and stimulation of single living cells under physiological conditions. A nanofluidic channel in the cantilever allows soluble molecules to be dispensed through a submicrometer aperture in the AFM tip. The sensitive AFM force feedback allows controlled approach of the tip to a sample for extremely local modification of surfaces in liquid environments. It also allows reliable discrimination between gentle contact with a cell membrane or its perforation. Using these two procedures, dyes have been introduced into individual living cells and even selected subcellular structures of these cells. The universality and versatility of the fluidFM will stimulate original experiments at the submicrometer scale not only in biology but also in physics, chemistry, and material science.

Scaling Hetero-Epitaxy from Layers to Three-Dimensional Crystals

Laying It on Thick The growth of one layered material onto a second lies at the heart of many electronic devices. However, if there is a lattice mismatch between the two materials, strains develop in the overgrowth material leading to bowing and cracking. Falub et al. (p. 1330; see the cover) patterned Si substrates into a series of pillars onto which they grew a germanium layer. The germanium initially coated the top of each silicon pillar but then widened as the layer thickened, leading to thick, crack-free germanium films. A space-filling array of self-limited three-dimensional epitaxial crystals averts wafer bowing, layer cracking, and dislocation propagation. Quantum structures made from epitaxial semiconductor layers have revolutionized our understanding of low-dimensional systems and are used for ultrafast transistors, semiconductor lasers, and detectors. Strain induced by different lattice parameters and thermal properties offers additional degrees of freedom for tailoring materials, but often at the expense of dislocation generation, wafer bowing, and cracks. We eliminated these drawbacks by fast, low-temperature epitaxial growth of Ge and SiGe crystals onto micrometer-scale tall pillars etched into Si(001) substrates. Faceted crystals were shown to be strain- and defect-free by x-ray diffraction, electron microscopy, and defect etching. They formed space-filling arrays up to tens of micrometers in height by a mechanism of self-limited lateral growth. The mechanism is explained by reduced surface diffusion and flux shielding by nearest-neighbor crystals.

Parallel AFM imaging and force spectroscopy using two‐dimensional probe arrays for applications in cell biology

Atomic force microscopy (AFM) investigations of living cells provide new information in both biology and medicine. However, slow cell dynamics and the need for statistically significant sample sizes mean that data collection can be an extremely lengthy process. We address this problem by parallelizing AFM experiments using a two‐dimensional cantilever array, instead of a single cantilever. We have developed an instrument able to operate a two‐dimensional cantilever array, to perform topographical and mechanical investigations in both air and liquid. Deflection readout for all cantilevers of the probe array is performed in parallel and online by interferometry. Probe arrays were microfabricated in silicon nitride. Proof‐of‐concept has been demonstrated by analyzing the topography of hard surfaces and fixed cells in parallel, and by performing parallel force spectroscopy on living cells. These results open new research opportunities in cell biology by measuring the adhesion and elastic properties of a large number of cells. Both properties are essential parameters for research in metastatic cancer development. Copyright

Unexpected Dominance of Vertical Dislocations in High-Misfit Ge/Si(001) Films and Their Elimination by Deep Substrate Patterning

An innovative strategy in dislocation analysis, based on comparison between continuous and tessellated film, demonstrates that vertical dislocations, extending straight up to the surface, easily dominate in thick Ge layers on Si(001) substrates. The complete elimination of dislocations is achieved by growing self-aligned and self-limited Ge microcrystals with fully faceted growth fronts, as demonstrated by AFM extensive etch-pit counts.

Fabrication of nanopore arrays and ultrathin silicon nitride membranes by block-copolymer-assisted lithography

Here we show a method for patterning a thin metal film using self-assembled block-copolymer micelles monolayers as a template. The obtained metallic mask is transferred by reactive ion etching in silicon oxide, silicon and silicon nitride substrates, thus fabricating arrays of hexagonally packed nanopores with tunable diameters, interspacing and aspect ratios. This technology is compatible with integration into a standard microtechnology sequence for wafer-scale fabrication of ultrathin silicon nitride nanoporous membranes with 80 nm mean pore diameter.

Piezoresistive cantilever array for life sciences applications

Atomic Force Microscopy (AFM) techniques are used with one- or two-dimensional arrays of piezoresistive probes for parallel imaging. We present a newly designed AFM platform to drive these passivated piezoresistive cantilever arrays in air and liquid environments. Large area imaging in liquid as well as qualitative and quantitative analysis of biological cells are demonstrated by the means of piezoresistive cantilever for the first time to our knowledge. Noise limitations in topography and force resolutions of these piezolevers are quantified.

Parallel scanning AFM with on-chip circuitry in CMOS technology

We present the first force sensors for application in Atomic Force Microscopy (AFM) fabricated with industrial CMOS technology. Sensing is based on two different detection schemes: a piezoresistive Wheatstone bridge and stress-sensing MOS transistors. The system combines on a single chip (i) two cantilevers for parallel scanning, (ii) thermal actuators for independent deflection of the two cantilevers, (iii) sensors to measure the deflection, and (iv) offset compensation and signal conditioning circuitry. The AFM probes were tested in contact and dynamic mode. In the dynamic mode, images with a resolution of better than 20 /spl Aring/ were recorded. Moreover, we successfully took parallel scanning images in contact mode.

Highly Mismatched, Dislocation-Free SiGe/Si Heterostructures

Defect-free mismatched heterostructures on Si substrates are produced by an innovative strategy. The strain relaxation is engineered to occur elastically rather than plastically by combining suitable substrate patterning and vertical crystal growth with compositional grading. Its validity is proven both experimentally and theoretically for the pivotal case of SiGe/Si(001).

Strain relaxation of GaAs/Ge crystals on patterned Si substrates

We report on the mask-less integration of GaAs crystals several microns in size on patterned Si substrates by metal organic vapor phase epitaxy. The lattice parameter mismatch is bridged by first growing 2-μm-tall intermediate Ge mesas on 8-μm-tall Si pillars by low-energy plasma enhanced chemical vapor deposition. We investigate the morphological evolution of the GaAs crystals towards full pyramids exhibiting energetically stable {111} facets with decreasing Si pillar size. The release of the strain induced by the mismatch of thermal expansion coefficients in the GaAs crystals has been studied by X-ray diffraction and photoluminescence measurements. The strain release mechanism is discussed within the framework of linear elasticity theory by Finite Element Method simulations, based on realistic geometries extracted from scanning electron microscopy images.

Transfer of Ultrasmall Iron Oxide Nanoparticles from Human Brain-Derived Endothelial Cells to Human Glioblastoma Cells

Nanoparticles (NPs) are being used or explored for the development of biomedical applications in diagnosis and therapy, including imaging and drug delivery. Therefore, reliable tools are needed to study the behavior of NPs in biological environment, in particular the transport of NPs across biological barriers, including the blood-brain tumor barrier (BBTB), a challenging question. Previous studies have addressed the translocation of NPs of various compositions across cell layers, mostly using only one type of cells. Using a coculture model of the human BBTB, consisting in human cerebral endothelial cells preloaded with ultrasmall superparamagnetic iron oxide nanoparticles (USPIO NPs) and unloaded human glioblastoma cells grown on each side of newly developed ultrathin permeable silicon nitride supports as a model of the human BBTB, we demonstrate for the first time the transfer of USPIO NPs from human brain-derived endothelial cells to glioblastoma cells. The reduced thickness of the permeable mechanical support compares better than commercially available polymeric supports to the thickness of the basement membrane of the cerebral vascular system. These results are the first report supporting the possibility that USPIO NPs could be directly transferred from endothelial cells to glioblastoma cells across a BBTB. Thus, the use of such ultrathin porous supports provides a new in vitro approach to study the delivery of nanotherapeutics to brain cancers. Our results also suggest a novel possibility for nanoparticles to deliver therapeutics to the brain using endothelial to neural cells transfer.