The monoclonal anti-dsRNA antibody J2 binds double-stranded RNAs (dsRNA) in an apparently sequence-nonspecific way. The mAb only recognizes antigens with double-stranded regions of at least 40 bp and its affinity to poly(A) poly(U) and to dsRNAs with mixed base pair composition is about tenfold higher than to poly(I) poly(C). Because no specific binding site could be determined, the number, the exact dimensions, and other distinct features of the binding sites on a given antigen are difficult to evaluate by biochemical methods. We therefore employed scanning force microscopy (SFM) as a method to analyze antibody-dsRNA interaction and protein-RNA binding in general. Several in vitro-synthesized dsRNA substrates, generated from the Dictyostelium PSV-A gene, were used. In addition to the expected sequence-nonspecific binding, imaging of the complexes indicated preferential binding of antibodies to the ends of dsRNA molecules as well as to certain internal sites. Analysis of 2,000 bound antibodies suggested that the consensus sequence of a preferential internal binding site is A 2 N 9 A 3 N 9 A 2 , thus presenting A residues on one face of the helix. The site was verified by site-directed mutagenesis, which abolished preferential binding to this region. The data demonstrate that SFM can be efficiently used to identify and characterize binding sites for proteins with no or incomplete sequence specificity. This is especially the case for many proteins involved in RNA metabolism.

The reproducible micromachining of hollow metal tips on Si cantilevers and their applicability to scanning probe microscopy techniques are described. Provided with apertures below 130 nm and hollow pyramidal tips proved to be highly suited probes for scanning near-field optical microscopy (SNOM). First results of combined SFM/SNOM measurements together with scanning electron microscopy (SEM) photographs of the new sensors are presented. The SNOM images show a resolution of about 100 nm demonstrating the usefulness of these probes.

Fast switching ‘transparent-to-black’ electrochromic devices are currently under investigation as potential candidates in modern applications like e-papers or with additional functionality as ultracompact iris or switchable neutral filter in camera systems. However, recent electrochromic devices show either a lack of contrast or slow response times. To overcome these deficiencies we focus on a careful material composition of the colouring hybrid electrodes in our device. We have established a nanoporous Sb-doped SnO electrode as supporting electrode for chemisorbed electrochromic tetraphenylbenzidine molecules due to its good conductivity in the redox potential range of the molecule. This hybrid electrode was combined with a modified nanoporous TiO / viologen electrode to realize a high performance, complementary electrochromic device. Fast switching time constants of 0.5 s and concurrently high change in optical density OD = 2.04 at 605 nm confirm our successful concept. The achieved colouration efficiency of 440 cm C exceeds every high contrast device presented so far.

The availability of very sharp, wear-proof, electrically conductive probes is one crucial issue for conductive atomic force microscopy (AFM) techniques such as scanning capacitance microscopy, scanning spreading resistance microscopy, and nanopotentiometry. The purpose of this systematic study is to give an overview of the existing probes and to evaluate their performance for the electrical techniques with emphasis on applications on Si at high contact forces. The suitability of the characterized probes has been demonstrated by applying conductive AFM techniques to test structures and state-of-the-art semiconductor devices. Two classes of probes were examined geometrically and electrically: Si sensors with a conductive coating and integrated pyramidal tips made of metal or diamond. Structural information about the conductive materials was obtained by electron microscopy and other analytical tools. Swift and nondestructive procedures to characterize the geometrical and electrical properties of the probes prior to the actual AFM experiment have been developed. Existing contact models have been used to explain variations in the electrical performance of the conductive probes.

Gold electrodes integrated into silicon scanning force microscopy (SFM) probes allow the acquisition of spatially correlated data for sample morphology (via SFM) and local electrochemical reactivity via scanning electrochemical microscopy (SECM). The lateral resolution of both techniques is controlled by different properties of the integrated probes. The topographic tracking provided by the SFM mechanism allows the realization of very small working distances for the SECM measurements. Microfabrication technology was used in order to reduce the size of the active electrode area of the tip into the sub-100 nm regime. The functionality of the probes was tested using electrochemical methods. Experiments revealed that the response could be quantitatively compared to numerical simulation. The low working distance, in combination with the small size of the active electrode area, allows for high lateral resolution in the SECM images. This is illustrated with different model substrates that cover a range of different rate constants and illustrate the dependence of the SECM contrast on the local kinetics of the sample in the sub-micrometre size range.