Evaluation of the hoof centre-of-pressure path in horses affected by chronic osteoarthritic pain.

INTRODUCTION: The Centre of Pressure (COP) is the single point summarising all forces transferred to the hoof during the stance phase of a stride. COP path (COPp) is the trajectory that COP follows from footstrike to lift-off. Aim of the present study was to characterize the COP and COPp in horses affected by osteoarthritis and chronic lameness. MATERIALS AND METHODS: Seventeen adult horses with a diagnosis of osteoarthritis and single limb chronic lameness were recruited. The COP was recorded using a wireless pressure measuring system (TekScan®) with sensors taped to the hooves (either fore- or hind limb, depending on lameness location). The COPp coordinates were further processed. Procrustes analysis was performed to assess the variability of single strides COPp and average COPp among strides, gaits, and limbs by calculating Procrustes distances (D-values). A linear mixed-effects model was run to analyse D-values differences for lame and sound limbs. Additionally, average COPp D-values and COPp hoofprint shape indices were compared for lame and sound limbs with the Signed Rank Test. RESULTS: At walk and trot the single-stride COPp D-values were significantly lower in lame than in sound limbs (marginal effects p<0.001). Analysis of the average COPp D-values confirmed that each hoof COPp is highly consistent with itself over subsequent trials but is different from the contralateral. COPp and hoofprint shape indices did not differ between sound and lame limbs. Footstrike and lift-off within the hoofprint showed that most horses had lateral footstrike and lift-off, independently of the lameness location. CONCLUSION: Our findings are in line with previous observations that COPp are highly repetitive and characteristic for each horse and limb. There seems to be a further decrease in COPp variability in the presence of a painful limb pathology.

PtyLab.m/py/jl: a cross-platform, open-source inverse modeling toolbox for conventional and Fourier ptychography.

Conventional (CP) and Fourier (FP) ptychography have emerged as versatile quantitative phase imaging techniques. While the main application cases for each technique are different, namely lens-less short wavelength imaging for CP and lens-based visible light imaging for FP, both methods share a common algorithmic ground. CP and FP have in part independently evolved to include experimentally robust forward models and inversion techniques. This separation has resulted in a plethora of algorithmic extensions, some of which have not crossed the boundary from one modality to the other. Here, we present an open source, cross-platform software, called PtyLab, enabling both CP and FP data analysis in a unified framework. With this framework, we aim to facilitate and accelerate cross-pollination between the two techniques. Moreover, the availability in Matlab, Python, and Julia will set a low barrier to enter each field.

Field-position dependent apodization in dark-field digital holographic microscopy for semiconductor metrology.

Measuring overlay between two layers of semiconductor devices is a crucial step during electronic chip fabrication. We present dark-field digital holographic microscopy that addresses various overlay metrology challenges that are encountered in the semiconductor industry. We present measurement results that show that the point-spread function of our microscope depends on the position in the field-of-view. We will show that this novel observation can be explained by a combination of the finite bandwidth of the light source and a wavelength-dependent focal length of the imaging lens. Moreover, we will also present additional experimental data that supports our theoretical understanding. Finally, we will propose solutions that reduce this effect to acceptable levels.

aPIE: an angle calibration algorithm for reflection ptychography.

Reflection ptychography is a lensfree microscopy technique particularly promising in regions of the electromagnetic spectrum where imaging optics are inefficient or not available. This is the case in tabletop extreme ultraviolet microscopy and grazing incidence small angle x ray scattering experiments. Combining such experimental configurations with ptychography requires accurate knowledge of the relative tilt between the sample and the detector in non-coplanar scattering geometries. Here, we describe an algorithm for tilt estimation in reflection ptychography. The method is verified experimentally, enabling sample tilt determination within a fraction of a degree. Furthermore, the angle-estimation uncertainty and reconstruction quality are studied for both smooth and highly structured beams.

Advances in laboratory-scale ptychography using high harmonic sources [Invited].

Extreme ultraviolet microscopy and wavefront sensing are key elements for next-generation ultrafast applications, such as chemically-resolved imaging, focal spot diagnostics in pump-and-probe experiments, and actinic metrology for the state-of-the-art lithography node at 13.5 nm wavelength. Ptychography offers a robust solution to the aforementioned challenges. Originally adapted by the electron and synchrotron communities, advances in the stability and brightness of high-harmonic tabletop sources have enabled the transfer of ptychography to the laboratory. This review covers the state of the art in tabletop ptychography with high harmonic generation sources. We consider hardware options such as illumination optics and detector concepts as well as algorithmic aspects in the analysis of multispectral ptychography data. Finally, we review technological application cases such as multispectral wavefront sensing, attosecond pulse characterization, and depth-resolved imaging.

Aberration calibration and correction with nano-scatterers in digital holographic microscopy for semiconductor metrology.

Overlay metrology measures pattern placement between two layers in a semiconductor chip. The continuous shrinking of device dimensions drives the need to explore novel optical overlay metrology concepts that can address many of the existing metrology challenges. We present a compact dark-field digital holographic microscope that uses only a single imaging lens. Our microscope offers several features that are beneficial for overlay metrology, like a large wavelength range. However, imaging with a single lens results in highly aberrated images. In this work, we present an aberration calibration and correction method using nano-sized point scatterers on a silicon substrate. Computational imaging techniques are used to recover the full wavefront error, and we use this to correct for the lens aberrations. We present measured data to verify the calibration method and we discuss potential calibration error sources that must be considered. A comparison with a ZEMAX calculation is also presented to evaluate the performance of the presented method.

Smartphone-Based Pelvic Movement Asymmetry Measures for Clinical Decision Making in Equine Lameness Assessment.

Visual evaluation of hindlimb lameness in the horse is challenging. Objective measurements, simultaneous to visual assessment, are used increasingly to aid clinical decision making. The aim of this study was to investigate the association of pelvic movement asymmetry with lameness scores (UK scale 0-10) of one experienced veterinarian. Absolute values of pelvic asymmetry measures, quantifying differences between vertical minima (AbPDMin), maxima (AbPDMax) and upward movement amplitudes (AbPDUp), were recorded during straight-line trot with a smartphone attached to the sacrum (n = 301 horses). Overall, there was a significant difference between lameness grades for all three asymmetry measures (p < 0.001). Five pair-wise differences (out of 10) were significant for AbPDMin (p ≤ 0.02) and seven for AbPDMax (p ≤ 0.03) and AbPDUp (p ≤ 0.02). Receiver operating curves assessed sensitivity and specificity of asymmetry measures against lameness scores. AbPDUp had the highest discriminative power (area under curve (AUC) = 0.801-0.852) followed by AbPDMax (AUC = 0.728-0.813) and AbPDMin (AUC = 0.688-0.785). Cut-off points between non-lame (grade 0) and lame horses (grades 1-4) with a minimum sensitivity of 75% were identified as AbPDUp ≥ 7.5 mm (67.6% specificity), AbPDMax ≥ 4.5 mm (51.9% specificity) and AbPDMin ≥ 2.5 mm (33.3% specificity). In conclusion, pelvic upward movement amplitude difference (AbPDUp) was the asymmetry parameter with the highest discriminative power in this study.

Monitoring of Organochlorine Pesticide and Polychlorinated Biphenyl Residues in Common Swifts (Apus apus) in the Region of Hannover, Lower Saxony, Germany.

The use of pesticides is associated with the decline of several avian species. In this study, we monitored the organochlorine contaminants in common swifts (Apus apus) in the years 2016 to 2018. These long-distance migrants breed in Europe and winter in Africa. Their only feeding source is aerial plankton. Pooled organ samples of 42 adult and 40 juvenile swifts were tested with the multi-residue method by gas chromatography-mass spectrometry (GC-TOF/MS). Predominantly, 4,4'-DDE, dieldrin, hexachlorobenzene (HCB), lindane and polychlorinated biphenyls (PCBs) were found in most of these common swifts. Only 4,4'-DDE (adult: 83 ± 70 µg/kg, juvenile: 17 ± 39 µg/kg) and dieldrin (adult: 2 ± 3 µg/kg, juvenile: 0.3 ± 1 µg/kg) concentrations were significantly different between adult and juvenile birds. All detected concentrations in our study were far lower than the previously recorded pesticide concentrations of common swifts in Italy and those which are known to cause toxicity and death in birds.

Ptychographic optical coherence tomography.

Ptychography is a robust computational imaging technique that can reconstruct complex light fields beyond conventional hardware limits. However, for many wide-field computational imaging techniques, including ptychography, depth sectioning remains a challenge. Here we demonstrate a high-resolution three-dimensional (3D) computational imaging approach, which combines ptychography with spectral-domain imaging, inspired by optical coherence tomography (OCT). This results in a flexible imaging system with the main advantages of OCT, such as depth-sectioning without sample rotation, decoupling of transverse and axial resolution, and a high axial resolution only determined by the source bandwidth. The interferometric reference needed in OCT is replaced by computational methods, simplifying hardware requirements. As ptychography is capable of deconvolving the illumination contributions in the observed signal, speckle-free images are obtained. We demonstrate the capabilities of ptychographic optical coherence tomography (POCT) by imaging an axially discrete lithographic structure and an axially continuous mouse brain sample.

Diffraction-based overlay metrology using angular-multiplexed acquisition of dark-field digital holograms.

In semiconductor device manufacturing, optical overlay metrology measures pattern placement between two layers in a chip with sub-nm precision. Continuous improvements in overlay metrology are needed to keep up with shrinking device dimensions in modern chips. We present first overlay metrology results using a novel off-axis dark-field digital holographic microscopy concept that acquires multiple holograms in parallel by angular multiplexing. We show that this concept reduces the impact of source intensity fluctuations on the noise in the measured overlay. With our setup we achieved an overlay reproducibility of 0.13 nm and measurements on overlay targets with known programmed overlay values showed good linearity of R2= 0.9993. Our data show potential for significant improvement and that digital holographic microscopy is a promising technique for future overlay metrology tools.

High-resolution microscopy through optically opaque media using ultrafast photoacoustics.

We present a high-resolution microscope capable of imaging buried structures through optically opaque materials with micrometer transverse resolution and a nanometer-scale depth sensitivity. The ability to image through such materials is made possible by the use of laser ultrasonic techniques, where an ultrafast laser pulse launches acoustic waves inside an opaque layer and subsequent acoustic echoes from buried interfaces are detected optically by a time-delayed probe pulse. We show that the high frequency of the generated ultrasound waves enables imaging with a transverse resolution only limited by the optical detection system. We present the imaging system and signal analysis and demonstrate its imaging capability on complex microstructured objects through 200 nm thick metal layers and gratings through 500 nm thickness. Furthermore, we characterize the obtained imaging performance, achieving a diffraction-limited transverse resolution of 1.2 µm and a depth sensitivity better than 10 nm.

Role of scattering by surface roughness in the photoacoustic detection of hidden micro-structures.

We present an experimental study in which we compare two different pump-probe setups to generate and detect high-frequency laser-induced ultrasound for the detection of gratings buried underneath optically opaque metal layers. One system is built around a high-fluence, low-repetition-rate femtosecond laser (1 kHz) and the other around a low-fluence, high-repetition-rate femtosecond laser (5.1 MHz). We find that the signal diffracted by the acoustic replica of the grating as a function of pump-probe time delay is very different for the two setups used. We attribute this difference to the presence of a constant background field due to optical scattering by interface roughness. In the low-fluence setup, the optical field diffracted by the acoustic replica is significantly weaker than the background optical field, with which it can destructively or constructively interfere. For the right phase difference between the optical fields, this can lead to a significant "amplification" of the weak field diffracted off the grating-shaped acoustic waves. For the high-fluence system, the situation is reversed because the field diffracted off the acoustic-wave-induced grating is significantly larger than the background optical field. Our measurements show that optical scattering by interface roughness must be taken into account to properly explain experiments on laser-induced ultrasound performed with high-repetition-rate laser systems and can be used to enhance signal strength.

Laser-induced ultrasonics for detection of low-amplitude grating through metal layers with finite roughness.

We report on the use of laser-induced ultrasonics for the detection of gratings with amplitudes as small as 0.5 nm, buried underneath an optically opaque nickel layer. In our experiments, we use gratings fabricated on top of a nickel layer on glass, and we optically pump and probe the sample from the glass side. The diffraction of the probe pulse from the acoustic echo from the buried grating is measured as a function of time. We use a numerical model to show how the various physical phenomena such as interface displacement, strain-optic effects, thermo-optic effects, and surface roughness influence the shape and strength of the time-dependent diffraction signal. More importantly, we use a Rayleigh-Rice scattering theory to quantify the amount of light scattering, which is then used as in input parameter in our numerical model to predict the time-dependent diffracted signal.

Impact of coherence length on the field of view in dark-field holographic microscopy for semiconductor metrology: theoretical and experimental comparisons.

Semiconductor manufacturers continue to increase the component densities on computer chips by reducing the device dimensions to less than 10 nm. This trend requires faster, more precise, and more robust optical metrology tools that contain complex and high-precision optics with challenging imaging requirements. Here, we present dark-field digital holographic microscopy as a promising optical metrology technique that uses optics with acceptable complexity. A theoretical analysis and an experimental demonstration of this technique are presented, showing the impact of the coherence length of the light source on the field of view. Finally, we also present the first holographically obtained images of metrology targets.

zPIE: an autofocusing algorithm for ptychography.

An autofocusing algorithm for ptychography is proposed. The method optimizes a sharpness metric that would be observed in a differential interference microscope and is valid for both amplitude and phase modulating specimens. We experimentally demonstrate that the algorithm, based on the extended ptychographic iterative engine (ePIE), calibrates the sample-detector distance with an accuracy within the depth of field of the ptychographic microscope. We show that the method can be used to determine slice separation in multislice ptychography, provided there are isolated regions on each slice of the specimen that do not axially overlap.

Measuring laser beam quality, wavefronts, and lens aberrations using ptychography.

We report on an approach for quantitative characterization of laser beam quality, wavefronts, and lens aberrations using ptychography with a near-infrared supercontinuum laser. Ptychography is shown to offer a powerful alternative for both beam propagation ratio M2 and wavefront measurements compared with existing techniques. In addition, ptychography is used to recover the transmission function of a microlens array for aberration analysis. The results demonstrate ptychography's flexibility in wavefront metrology and optical shop testing.

Generation and characterization of focused helical x-ray beams.

The phenomenon of orbital angular momentum (OAM) affects a variety of important applications in visible optics, including optical tweezers, free-space communication, and 3D localization for fluorescence imaging. The lack of suitable wavefront shaping optics such as spatial light modulators has inhibited the ability to impart OAM on x-ray and electron radiation in a controlled way. Here, we report the experimental observation of helical soft x-ray beams generated by holographically designed diffractive optical elements. We demonstrate that these beams rotate as a function of propagation distance and measure their vorticity and coherent mode structure using ptychography. Our results establish an approach for controlling and shaping of complex focused beams for short wavelength scanning microscopy and OAM-driven applications.

Optical parametric chirped pulse amplifier producing ultrashort 10.5 mJ pulses at 1.55 µm.

We present an optical parametric chirped pulse amplifier (OPCPA) delivering 10.5 mJ pulses with durations down to 220 fs, at 100 Hz repetition rate, centered at 1550 nm. The system is pumped by a picosecond Nd:YAG amplifier at 1064 nm based on quasi-continuous-wave diode pumping and seeded by a femtosecond mode-locked Er fiber laser at 1550 nm. This choice of wavelengths enables the use of well-established technology and optical components for both pump and signal beams, resulting in a straightforward and robust system design and the ability for further power scaling to be used in high-energy laser-produced plasma experiments.

Interference probe ptychography for computational amplitude and phase microscopy.

We have developed an approach to Fresnel domain ptychography in which the illumination consists of an interference pattern. This pattern is conveniently created by overlapping two coherent beams at an angle. Only the phase and orientation of the interferometric fringe pattern needs to be scanned to reconstruct a high-fidelity object image, which alleviates the requirements for accurate sample positioning and system stability. As such, the resulting imaging systems can be constructed in an extremely simple and robust way. Object images are reconstructed from recorded Fresnel diffraction data using a modified ptychographical iterative engine. We demonstrate the capabilities of this imaging system by recording images of various biological samples, demonstrating quantitative phase contrast as well as a spatial resolution better than 2.2 µm.

Fourier transform holography with extended references using a coherent ultra-broadband light source.

We demonstrate a technique that enables lensless holographic imaging with extended reference structures, using ultra-broadband radiation sources for illumination. We show that this 'two-pulse imaging' approach works with one- and two-dimensional HERALDO reference structures, and demonstrate that the obtained spectrally resolved data can be used to improve the signal-to-noise ratio in the final image. Intensity stitching of multiple exposures is applied to increase the detected dynamic range, leading to an improved image reconstruction. Furthermore, we show that a combination of holography and iterative phase retrieval can be used to obtain high-quality images quickly and reliably, by using the HERALDO reconstruction as the initial support constraint in the iterative phase retrieval algorithm. A signal-to-noise improvement of two orders of magnitude is achieved compared to the basic HERALDO result.