K.O. van der Werf

Combined AFM and confocal fluorescence microscope for applications in bio‐nanotechnology

We present a custom‐designed atomic force fluorescence microscope (AFFM), which can perform simultaneous optical and topographic measurements with single molecule sensitivity throughout the whole visible to near‐infrared spectral region. Integration of atomic force microscopy (AFM) and confocal fluorescence microscopy combines the high‐resolution topographical imaging of AFM with the reliable (bio)‐chemical identification capability of optical methods. The AFFM is equipped with a spectrograph enabling combined topographic and fluorescence spectral imaging, which significantly enhances discrimination of spectroscopically distinct objects. The modular design allows easy switching between different modes of operation such as tip‐scanning, sample‐scanning or mechanical manipulation, all of which are combined with synchronous optical detection. We demonstrate that coupling the AFM with the fluorescence microscope does not compromise its ability to image with a high spatial resolution. Examples of several modes of operation of the AFFM are shown using two‐dimensional crystals and membranes containing light‐harvesting complexes from the photosynthetic bacterium Rhodobacter sphaeroides.

Mechanical testing of electrospun PCL fibers

Poly(ε-caprolactone) (PCL) fibers ranging from 250 to 700 nm in diameter were produced by electrospinning a polymer tetrahydrofuran/N,N-dimethylformamide solution. The mechanical properties of the fibrous scaffolds and individual fibers were measured by different methods. The Youngs moduli of the scaffolds were determined using macro-tensile testing equipment, whereas single fibers were mechanically tested using a nanoscale three-point bending method, based on atomic force microscopy and force spectroscopy analyses. The modulus obtained by tensile-testing eight different fiber scaffolds was 3.8±0.8 MPa. Assuming that PCL fibers can be described by the bending model of isotropic materials, a Youngs modulus of 3.7±0.7 GPa was determined for single fibers. The difference of three orders of magnitude observed in the moduli of fiber scaffolds vs. single fibers can be explained by the lacunar and random structure of the scaffolds.

A new imaging mode in Atomic Force Microscopy based on the error signal

A new imaging mode, the error signal mode, is introduced to atomic force microscopy. In this mode, the error signal is displayed while imaging in the height mode. The feedback loop serves as a high-pass filter that filters out the low spatial frequency components of the surface, leaving only the high spatial frequency components of the surface to contribute to the error signal and to be displayed. At a scan rate of typically 10 lines per second, images taken in this mode show very fine detail. Since the applied force stays nearly constant, the error signal mode is especially suitable for imaging soft biological samples with a high level of detail without damaging the surface.

Nano-mechanical tuning and imaging of a photonic crystal micro-cavity resonance

We show that nano-mechanical interaction using atomic force microscopy (AFM) can be used to map out mode-patterns of an optical micro-resonator with high spatial accuracy. Furthermore we demonstrate how the Q-factor and center wavelength of such resonances can be sensitively modified by both horizontal and vertical displacement of an AFM tip consisting of either Si(3)N(4) or Si material. With a silicon tip we are able to tune the resonance wavelength by 2.3 nm, and to set Q between values of 615 and zero, by expedient positioning of the AFM tip. We find full on/off switching for less than 100 nm vertical, and for 500 nm lateral displacement at the strongest resonance antinode locations.

Tuning fork shear-force feedback

Investigations have been performed on the dynamics of a distance regulation system based on an oscillating probe at resonance. This was examined at a tuning fork shear-force feedback system, which is used as a distance control mechanism in near-field scanning optical microscopy. In this form of microscopy, a tapered optical fiber is attached to the tuning fork and scanned over the sample surface to be imaged. Experiments were performed measuring both amplitude and phase of the oscillation of the tuning fork as a function of driving frequency and tip-sample distance. These experiments reveal that the resonance frequency of the tuning fork changes upon approaching the sample. Both the amplitude and the phase of the tuning fork can be used as distance control parameter in the feedback system. Using the amplitude a second-order behavior is observed, while with phase only a first-order behavior is observed. Numerical calculations confirm these observations. This first-order behavior results in an improved stability of the feedback system. As an example, a sample consisting of DNA strands on mica was imaged which showed the height of the DNA as 1.4 +/- 0.2 nm.

Automatic kelvin probe compatible with ultrahigh vacuum

This article describes a new type of in situ ultrahigh‐vacuum compatible kelvin probe based on a voice‐coil driving mechanism. This design exhibits several advantages over conventional mechanical feed‐through and (in situ) piezoelectric devices in regard to the possibility of multiple probe geometry, flexibility of probe geometry, amplitude of oscillation, and pure parallel vibration. Automatic setup and constant spacing features are achieved using a digital‐to‐analog converter (DAC) steered offset potential. The combination of very low driver noise pick‐up and data‐acquisition system (DAS) signal processing techniques results in a work function (wf  ) resolution, under optimal conditions, of <0.1 meV. Due to its high surface sensitivity and compatibility with standard sample cleaning and analysis techniques this design has numerous applications in surface studies, e.g., adsorption kinetics, sample topography and homogeneity, sputter profiles, etc. For semiconductor specimens the high wf resolution makes it eminently suitable for surface photovoltage (SPV) spectroscopy.

Micromechanical analysis of native and cross-linked collagen type I fibrils supports the existence of microfibrils

The mechanical properties of individual collagen fibrils of approximately 200 nm in diameter were determined using a slightly adapted AFM system. Single collagen fibrils immersed in PBS buffer were attached between an AFM cantilever and a glass surface to perform tensile tests at different strain rates and stress relaxation measurements. The stress-strain behavior of collagen fibrils immersed in PBS buffer comprises a toe region up to a stress of 5 MPa, followed by the heel and linear region at higher stresses. Hysteresis and strain-rate dependent stress-strain behavior of collagen fibrils were observed, which suggest that single collagen fibrils have viscoelastic properties. The stress relaxation process of individual collagen fibrils could be best fitted using a two-term Prony series. Furthermore, the influence of different cross-linking agents on the mechanical properties of single collagen fibrils was investigated. Based on these results, we propose that sliding of microfibrils with respect to each other plays a role in the viscoelastic behavior of collagen fibrils in addition to the sliding of collagen molecules with respect to each other. Our finding provides a better insight into the relationship between the structure and mechanical properties of collagen and the micro-mechanical behavior of tissues.

Operation of a scanning near-field optical microscope in reflection in combination with a scanning force microscope

Images obtained with a scanning near field optical microscope (SNOM) operating in reflection are presented. We have obtained the first results with a SiN tip as optical probe. The instrument is simultaneously operated as a scanning force microscope (SFM). Moreover, the instrument incorporates an inverted light microscope (LM) for preselection of a scan area. The SiN probe is operated in the contact regime causing a highly improved lateral resolution in the optical image compared to an alternative set-up using a fiber probe, which is also presented. The combined microscope is operated either in open loop or as a force regulated SNOM. Near field optical images can be directly compared with the topography displayed in the simultaneously recorded SFM image.

Dependence of silicon position-detector bandwidth on wavelength, power, and bias

We have developed a two-LED wobbler system to generate the spatial displacement of total light intensity on a detector surface, facilitating the acquisition of frequency responses up to 600 kHz with high accuracy. We have used this setup to characterize the low-pass filtering behavior of silicon-based position detectors for wavelengths above 850 nm by acquiring the frequency responses of several quadrant detectors and position-sensitive detectors as functions of wavelength, applied bias voltage, and total light power. We observed an increase in bandwidth for an increase in applied bias voltage and incident-light intensity. The combined effect of these parameters is strongly dependent on the detector used and has significant implications for the use of these detectors in scanning probe and optical tweezers applications.

Integrated automatic modular measuring system

This paper describes a versatile automatic measuring system composed of discrete modules. The modules can operate in both stand‐alone and remote modes and are interconnected using an IEEE‐488 bus, allowing utilization of standard measurement apparatus and peripherals. The system design allows user optimization of the measurement procedure, which can thus be tailored to meet specific experimental requirements. The flexibility of this system is demonstrated by its application in spectroscopic ellipsometry.