Nektarios Koukourakis

Axial scanning in confocal microscopy employing adaptive lenses (CAL)

In this paper we analyze the capability of adaptive lenses to replace mechanical axial scanning in confocal microscopy. The adaptive approach promises to achieve high scan rates in a rather simple implementation. This may open up new applications in biomedical imaging or surface analysis in micro- and nanoelectronics, where currently the axial scan rates and the flexibility at the scan process are the limiting factors. The results show that fast and adaptive axial scanning is possible using electrically tunable lenses but the performance degrades during the scan. This is due to defocus and spherical aberrations introduced to the system by tuning of the adaptive lens. These detune the observation plane away from the best focus which strongly deteriorates the axial resolution by a factor of ~2.4. Introducing balancing aberrations allows addressing these influences. The presented approach is based on the employment of a second adaptive lens, located in the detection path. It enables shifting the observation plane back to the best focus position and thus creating axial scans with homogeneous axial resolution. We present simulated and experimental proof-of-principle results.

Photorefractive two-wave mixing for image amplification in digital holography

We use photorefractive two-wave mixing for coherent amplification of the object beam in digital holographic recording. Both amplitude and phase reconstruction benefit from the prior amplification as they have an increased SNR. We experimentally verify that the amplification process does not affect the phase of the wavefield. This allows for digital holographic phase analysis after amplification. As the grating formation in photorefractive crystals is just driven by coherent light, the crystal works as a coherence gate. Thus the proposed combination allows for applying digital holography for imaging through scattering media, after the image bearing light is coherence gated and filtered out of scattered background. We show experimental proof-of principle results.

Integrity of Micro-Hotplates During High-Temperature Operation Monitored by Digital Holographic Microscopy

An investigation on the integrity of micro-hotplates using in situ digital holographic microscopy is reported. The surface topography and surface evolution of the devices during high-temperature operation (heating/cooling cycles) is measured with nanometer-scale resolution. A localized permanent out-of-plane surface deformation of 40% of the membrane thickness caused by the top measurement electrodes occurring after the first cycle is observed. The integrity-related issues caused by such a permanent deformation are discussed.

Volumetric HiLo microscopy employing an electrically tunable lens.

Electrically tunable lenses exhibit strong potential for fast motion-free axial scanning in a variety of microscopes. However, they also lead to a degradation of the achievable resolution because of aberrations and misalignment between illumination and detection optics that are induced by the scan itself. Additionally, the typically nonlinear relation between actuation voltage and axial displacement leads to over- or under-sampled frame acquisition in most microscopic techniques because of their static depth-of-field. To overcome these limitations, we present an Adaptive-Lens-High-and-Low-frequency (AL-HiLo) microscope that enables volumetric measurements employing an electrically tunable lens. By using speckle-patterned illumination, we ensure stability against aberrations of the electrically tunable lens. Its depth-of-field can be adjusted a-posteriori and hence enables to create flexible scans, which compensates for irregular axial measurement positions. The adaptive HiLo microscope provides an axial scanning range of 1 mm with an axial resolution of about 4 μm and sub-micron lateral resolution over the full scanning range. Proof of concept measurements at home-built specimens as well as zebrafish embryos with reporter gene-driven fluorescence in the thyroid gland are shown.

Wavefront shaping for imaging-based flow velocity measurements through distortions using a Fresnel guide star

Imaging-based flow measurement techniques, like particle image velocimetry (PIV), are vulnerable to time-varying distortions like refractive index inhomogeneities or fluctuating phase boundaries. Such distortions strongly increase the velocity error, as the position assignment of the tracer particles and the decrease of image contrast exhibit significant uncertainties. We demonstrate that wavefront shaping based on spatially distributed guide stars has the potential to significantly reduce the measurement uncertainty. Proof of concept experiments show an improvement by more than one order of magnitude. Possible applications for the wavefront shaping PIV range from measurements in jets and film flows to biomedical applications.

Depth-filtered digital holography

We introduce depth-filtered digital holography (DFDH) as a method for quantitative tomographic phase imaging of buried layers in multilayer samples. The procedure is based on the acquisition of multiple holograms for different wavelengths. Analyzing the intensity over wavelength pixel wise and using an inverse Fourier transform leads to a depth-profile of the multilayered sample. Applying a windowed Fourier transform with a narrow window, we choose a depth-of interest (DOI) which is used to synthesize filtered interference patterns that just contain information of this limited depth. We use the angular spectrum method to introduce an additional spatial filtering and to reconstruct the corresponding holograms. After a short theoretical framework we show experimental proof-of-principle results for the method.

High room-temperature optical gain in Ga(NAsP)/Si heterostructures

We analyze the modal gain of Ga(NAsP) multi quantum-well heterostructures pseudomorphically grown on (001) silicon substrate by metal-organic vapor-phase epitaxy. Using the variable stripe length method, we obtain high modal gain values up to 78 cm−1 at room temperature that are comparable to the values of common high quality III-V laser material. We find good agreement between experimental results and theoretically calculated gain spectra obtained using a microscopic model. The results underline the high potential of Ga(NAsP) as an active material for directly electrically pumped lasers on silicon substrate.

On the speckle number of interferometric velocity and distance measurements of moving rough surfaces.

The minimum achievable systematic uncertainty of interferometric measurements is fundamentally limited due to speckle noise. Numerical and physical experiments, regarding the achievable measurement uncertainty of Mach-Zehnder based velocity and position sensors, are presented at the example of the laser Doppler distance sensor with phase evaluation. The results show that the measurement uncertainty depends on the number of speckles on the photo detectors. However, while the systematic uncertainty due to the speckle effect decreases, the random uncertainty due to noise from the photo detector increases with increasing speckle number. This results in a minimal total measurement uncertainty for an optimal speckle number on the photo detector, which is achieved by adjusting the aperture of the detection optics.

Multiwavelength phase unwrapping and aberration correction using depth filtered digital holography

In this Letter, we present a new approach to processing data from a standard spectral domain optical coherence tomography (OCT) system using depth filtered digital holography (DFDH). Intensity-based OCT processing has an axial resolution of the order of a few micrometers. When the phase information is used to obtain optical path length differences, subwavelength accuracy can be achieved, but this limits the resolvable step heights to half of the wavelength of the system. Thus there is a metrology gap between phase- and intensity-based methods. Our concept addresses this metrology gap by combining DFHD with multiwavelength phase unwrapping. Additionally, the measurements are corrected for aberrations. Here, we present proof of concept measurements of a structured semiconductor sample.

Single-shot holography for depth resolved three dimensional imaging

We introduce a method for depth-resolved photorefractive holographic imaging with potentially extremely short acquisition time for a complete three dimensional (3D) image. By combining the advantages of full-field frequency-domain optical coherence tomography with those of photorefractive holography our concept is capable of obtaining 3D information with only one single shot. We describe the operation principle of our concept and give a first experimental proof of principle.