Volcano-Shaped Scanning Probe Microscopy Probe for Combined Force-Electrogram Recordings from Excitable Cells

Atomic force microscopy based approaches have led to remarkable advances in the field of mechanobiology. However, linking the mechanical cues to biological responses requires complementary techniques capable of recording these physiological characteristics. In this study, we present an instrument for combined optical, force, and electrical measurements based on a novel type of scanning probe microscopy cantilever composed of a protruding volcano-shaped nanopattemed microelectrode (nanovolcano probe) at the tip of a suspended microcantilever. This probe enables simultaneous force and electrical recordings from single cells. Successful impedance measurements on mechanically stimulated neonatal rat cardiomyocytes in situ were achieved using these nanovolcano probes. Furthermore, proof of concept experiments demonstrated that extracellular field potentials (electrogram) together with contraction displacement curves could simultaneously be recorded. These features render the nanovolcano probe especially suited for mechanobiological studies aiming at linking mechanical stimuli to electrophysiological responses of single cells.

New photoplastic fabrication techniques and devices based on high aspect ratio photoresist

Grégoire Genolet

This thesis deals with the development of microfabrication technologies based on photoplastic structuring by lithographic and molding techniques. These technologies, combined with an original releasing method, allow for the simple fabrication of pseudo three-dimensional, soft, photoplastic microstructures with features and shapes that are difficult to obtain with standard micro-machining. The photoplastic material used is the SU-8 photoresist, a material that is used increasingly in the growing field of micro-electro-mechanical systems (MEMS). Photoplastic SU-8 microstructure fabrication is based on a combination of multi-layer spin-coating, molding and photolithographic processing of the resist on a prestructured silicon substrate. The final product is obtained by release of the structure from the substrate. This thesis describes several SU-8 microstructures which have been developed with an emphasis on atomic force microscopy (AFM) and scanning-near field optical microscopy (SNOM) probes. Scanning probe microscopy is a well-established technique for surface analysis, but batch-fabricated, low-cost probes still remain a challenging issue. Using a polymer for the cantilever facilitates the realization of mechanical properties that are difficult to achieve with classical silicon technology. The design, fabrication and testing of single lever and cassettes of multiple single-lever probes are presented and demonstrates the potential for fabrication structures with complicated shape and features. The fabrication process for SU-8 AFM probes is a simple batch process in which the integrated tips and the levers are defined in one photolithography step. Tip radii of curvature smaller than 15 nm have been obtained. The cantilever thickness depends on the spin-coating parameters, hence it can be very well controlled over a full wafer. Photoplastic cantilevers with thicknesses ranging from 1 to 6 μm have been produced. Imaging soft, condensed matter with photo-plastic levers, which uses laser beam deflection sensing, exhibits a resolution that compares well with that of commercially available silicon cantilevers. Lateral resolution of 5–6 nm has been estimated from imaging DNA-plasmid molecules. A vertical resolution of the order of 0.1 nm has been found. A similar fabrication technique was also developed to fabricate photo-plastic tips for SNOM that are to be attached to optical fibers. This technique allows optical apertures to be integrated at the end of the well-defined tip directly by probe fabrication, without the need for any post-processing for the aperture formation. Sub-100 nm aperture have been fabricated using this technique. Simple fabrication, as well as topographical and optical imaging demonstrate the potential of photoplastic-based probes for both AFM and SNOM applications, as well as for future combined probes development. In addition, the fabrication of functional microstructures by using SU-8 processing needs to be combined with other microfabrication techniques. Simple releasing of the molded structures from the substrate is especially of great importance. A sacrificial layer technique based on electrochemical etching enhancement has been developed and combined with the fabrication of the different photoplastic SU-8 probes presented. This technique allows the fast releasing of large microstructures and has been demonstrated by releasing other SU-8 photoplastic microfabricated devices.

EPFL2001

Force-distance curves by atomic force microscopy

Giovanni Dietler

Atomic force microscopy (AFM) force-distance curves have become a fundamental tool in several fields of research, such as surface science, materials engineering, biochemistry and biology. Furthermore, they have great importance for the study of surface interactions from a theoretical point of view. Force-distance curves have been employed for the study of numerous materials properties and for the characterization of all the known kinds of surface forces. Since 1989, several techniques of acquisition and analysis have arisen. An increasing number of systems, presenting new kinds of forces, have been analyzed. AFM force-distance curves are routinely used in several kinds of measurement, for the determination of elasticity, Hamaker constants, surface charge densities, and degrees of hydrophobicity. The present review is designed to indicate the theoretical background of AFM force-distance curves as well as to present the great variety of measurements that can be performed with this tool. Section 1 is a general introduction to AFM force-distance curves. In Sections 2-4 the fundamentals of the theories concerning the three regions of force-distance curves are summarized. In particular, Section 2 contains a review of the techniques employed for the characterization of the elastic properties of materials. After an overview of calibration problems (Section 5), the different forces that can be measured with AFM force-distance curves are discussed. Capillary, Coulomb, Van der Waals, double-layer, solvation, hydration, hydrophobic, specific and steric forces are considered. For each force the available theoretical aspects necessary for the comprehension of the experiments are provided. The main experiments concerning the measurements of such forces are listed, pointing out the experimental problems, the artifacts that are likely to affect the measurement, and the main established results. Experiments up to June 1998 are reviewed. Finally, in Section 7, techniques to acquire force-distance curves sequentially and to draw bidimensional maps of different parameters are listed.

Elsevier1999

Study of probes and substrates for low temperature atomic force microscopy and biological applications

Giovanni Dietler, Aleksandra Radenovic

The atomic force microscopy in ultrahigh vacuum and at low temperature demonstrated its excellent capability to reach atomic resolution. Nevertheless in the case of biological samples high resolution has been achieved only in very few cases. We demonstrated here the importance of the appropriate choice of probes and substrates in order to image DNA at low temperature with high resolution. We investigated properties of three types of cantilevers and they were studied by scanning electron microscopy as a function of temperature. A large bending of cantilevers, which were coated from both sides, was observed at low temperatures. Therefore uncoated cantilevers pare strongly recommended for low temperature applications. Different methods for immobilization of DNA on the substrate are examined at low temperatures. First images of linear DNA on graphite at 82 K under ultrahigh vacuum conditions are presented.