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The parallel-plate compression test is one of the simplest ways to measure the mechanical properties of a material. In this test, the Young’s modulus ( E ) and the Poisson’s ratio ( ν ) of the material are determined directly without applying any additional modelling and parameter fitting in the post-processing. This is, however, limited when dealing soft biological materials due to their inherent properties such as being inhomogeneous, microscopic, and overly compliant. By combining an interferometry-assisted parallel-plate compression system and a confocal microscope, we were able to overcome these limitations and measure the E (315 ± 52 Pa) and ν (0.210 ± 0.043) of fixated and permeabilized bovine oocytes.

We propose a new type of self-referencing and multiplexed refractive index (RI) sensor based on a compound optical microresonator structure consisting of Fabry-Pérot (FP) resonators coupled with microring resonators. The transmission spectra shows resonant features that are superimposed on a background defined by FP oscillations. The resonances have asymmetric Fano-like non-Lorentzian shapes, which are used as sensing peaks, while the FP oscillations are used as reference peaks for internal self-referencing. The sensing peaks shift linearly with the increased RI of the cladding in the microring resonator, while FP peaks stay constant. When the temperature is increased, both the FP peaks and the Fano resonances shift linearly at the same rate, which eliminates the temperature effect on RI measurements. We theoretically analyzed that the two-mirror FP resonator coupled with a single microring resonator and optimized its sensing performance through finite-difference time-domain simulations. A sensitivity value of 220 nm/RIU and a maximum figure of merit of 4400 RIU-1 were achieved. We also proposed two possible multiplexing schemes consisting of two-mirror and three-mirror FP resonators coupled with two microring resonators of different radii. The proposed sensor concept is simple, easy-to-fabricate, self-calibrating and can be used for simultaneous measurements of different samples.

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.

Quantitative velocity estimations in optical coherence tomography requires the estimation of the axial and lateral flow components. Optical coherence tomography measures the depth resolved complex field reflected from a sample. While the axial velocity component can be determined from the Doppler shift or phase shift between a pair of consecutive measurements at the same location, the estimation of the lateral component for in vivo applications is still challenging. One approach to determine lateral velocity is multiple simultaneous measurements at different angles. In another approach the lateral component can be retrieved through repeated measurements at (nearly) the same location by an analysis of the decorrelation over time. In this paper we follow the latter approach. We describe a model for the complex field changes between consecutive measurements and use it to predict the uncertainties for amplitude-based, phase-based and complex algorithms. The uncertainty of the flow estimations follows from a statistical analysis and is determined by the number of available measurements and the applied analysis method. The model is verified in phantom measurements and the dynamic range of velocity estimations is investigated. We demonstrate that phase-based and complex (phasor) based lateral flow estimation methods are superior to amplitude-based algorithms.

Light microscopes don't have the ability to identify silicone (PDMS) particles from implants in histological slides. We describe a label-free method based on stimulated Raman scattering microscopy to locate and identify silicone in breast tissue.

Polarization-sensitive optical coherence tomography has been used to image two healthy and four diseased lungs ex vivo together with histology. Differences between lungs were found in alveoli size and airway smooth muscle thickness.

Retinal motion detection with an accuracy of 0.77 arcmin corresponding to 3.7 µm on the retina is demonstrated with a novel digital micromirror device based ophthalmoscope. By generating a confocal image as a reference, eye motion could be measured from consecutively measured subsampled frames. The subsampled frames provide 7.7 millisecond snapshots of the retina without motion artifacts between the image points of the subsampled frame, distributed over the full field of view. An ophthalmoscope pattern projection speed of 130 Hz enabled a motion detection bandwidth of 65 Hz. A model eye with a scanning mirror was built to test the performance of the motion detection algorithm. Furthermore, an in vivo motion trace was obtained from a healthy volunteer. The obtained eye motion trace clearly shows the three main types of fixational eye movements. Lastly, the obtained eye motion trace was used to correct for the eye motion in consecutively obtained subsampled frames to produce an averaged confocal image correct for motion artefacts.

Three-dimensional confocal fluorescence imaging of in vivo tissues is challenging due to sample motion and limited imaging speeds. In this paper a novel method is therefore presented for scanning confocal epi-fluorescence microscopy with instantaneous depth-sensing based on self-interference fluorescence microscopy (SIFM). A tabletop epi-fluorescence SIFM setup was constructed with an annular phase plate in the emission path to create a spectral self-interference signal that is phase-dependent on the axial position of a fluorescent sample. A Mach-Zehnder interferometer based on a 3 × 3 fiber-coupler was developed for a sensitive phase analysis of the SIFM signal with three photon-counter detectors instead of a spectrometer. The Mach-Zehnder interferometer created three intensity signals that alternately oscillated as a function of the SIFM spectral phase and therefore encoded directly for the axial sample position. Controlled axial translation of fluorescent microsphere layers showed a linear dependence of the SIFM spectral phase with sample depth over axial image ranges of 500 µm and 80 µm (3.9 × Rayleigh range) for 4 × and 10 × microscope objectives respectively. In addition, SIFM was in good agreement with optical coherence tomography depth measurements on a sample with indocyanine green dye filled capillaries placed at multiple depths. High-resolution SIFM imaging applications are demonstrated for fluorescence angiography on a dye-filled capillary blood vessel phantom and for autofluorescence imaging on an ex vivo fly eye.

Single pixel imaging can be the preferred method over traditional 2D-array imaging in spectral ranges where conventional cameras are not available. However, when it comes to real-time video imaging, single pixel imaging cannot compete with the framerates of conventional cameras, especially when high-resolution images are desired. Here we evaluate the performance of an imaging approach using two detectors simultaneously. First, we present theoretical results on how low SNR affects final image quality followed by experimentally determined results. Obtained video framerates were doubled compared to state of the art systems, resulting in a framerate from 22 Hz for a 32 × 32 resolution to 0.75 Hz for a 128 × 128 resolution image. Additionally, the two detector imaging technique enables the acquisition of images with a resolution of 256 × 256 in less than 3 s.

We demonstrate a compact, wide-field, quantitative phase contrast microscope that does not require lenses for image formation. High-resolution images are retrieved from Fresnel diffraction patterns recorded at multiple wavelengths, combined with a robust iterative phase retrieval algorithm. Quantitative phase contrast images of living cultured neurons are obtained with a transverse resolution of

We demonstrate a compact lensless microscope which can capture video-rate phase contrast images of moving objects and allows numerical scanning of the focal distance after recording. Using only an RGB-detector and illumination from a single mode fiber, diffraction patterns at three wavelengths are recorded simultaneously, enabling high-speed data collection and reconstruction of phase and amplitude. The technique is used for imaging of a moving test target, beads in a flow cell, and imaging of Caenorhabditis elegans moving in a droplet of liquid.

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.

We report on the development of the first immersion photoacoustic spectrometer (iPAS) for arcing fault detection in power transformers. The spectrometer consists of a detection system and an all-optical photoacoustic sensing head mounted inside a small permeable chamber where dissolved C2H2 diffuses while the transformer oil is kept out. Our all-optical iPAS sensor can be placed directly inside an oil bath and measure dissolved C2H2 with the sensitivity and linearity needed for in situ arcing fault detection. Moreover, its fast response time holds great promise for extra-early fault diagnosis.

Scattering prevents light from being focused in turbid media. The effect of scattering can be negated through wavefront shaping techniques when a localized form of feedback is available. Even in the absence of such a localized reporter, wavefront shaping can blindly form a single diffraction-limited focus when the feedback response is nonlinear. We developed and experimentally validated a model that accurately describes the statistics of this blind focusing process. We show that maximizing the nonlinear feedback signal does not always result in the formation of a focus. Using our model, we can calculate the minimal requirements to blindly focus light through strongly scattering media., Comment: 13 pages, 4 figures

Optical coherence tomography (OCT) is now a well-established modality for high-resolution cross-sectional and three-dimensional imaging of transparent and translucent samples and tissues. Conventional, intensity based OCT, however, does not provide a tissue-specific contrast, causing an ambiguity with image interpretation in several cases. Polarization sensitive (PS) OCT draws advantage from the fact that several materials and tissues can change the light’s polarization state, adding an additional contrast channel and providing quantitative information. In this paper, we review basic and advanced methods of PS-OCT and demonstrate its use in selected biomedical applications.

Taking inspiration from conventional top-down micromachining techniques, we have fabricated a low mass gold fiber-top cantilever via align-and-shine photolithography. The cantilever is characterized by measuring its resonance frequency and mechanical quality factor. Our results show that the device grants mass sensitivity comparable to that reported for similar standard cantilevers. This proof-of-concept paves the way to series production of highly sensitive fiber-top devices for remote detection of biochemical substances.

We present a digital spectral shaping technique to reduce the sidelobes (ringing) of the axial point-spread function in optical coherence tomography for non-Gaussian-shaped source spectra. The spectra of two superluminescent diodes were combined to generate a spectrum with significant modulation. Images of onion cells demonstrate the improved image quality in a turbid biological sample. A quantitative analysis of the accompanying penalty in signal-to-noise ratio is given. (C) 2002 Optical Society of America.

A high-speed single-mode fiber-based polarization-sensitive optical coherence tomography (PS OCT) system was developed. With a polarization modulator, Stokes parameters of reflected flight for four input polarization states are measured as a function of depth. A phase modulator in the reference arm of a Michelson interferometer permits independent control of the axial scan rate and carrier frequency. In vivo PS OCT images of human skin are presented, showing subsurface structures that are not discernible in conventional OCT images. A phase retardation image in tissue is calculated based on the reflected Stokes parameters of the four input polarization states. (C) 2000 Optical Society of America.

The invention is a fast-scanning ODT system that uses phase information derived from a Hilbert transformation to increase the sensitivity of flow velocity measurements while maintaining high spatial resolution. The significant increases in scanning speed and velocity sensitivity realized by the invention make it possible to image in vivo blood flow in human skin. The method of the invention overcomes the inherent limitations of the prior art ODT by using a phase change between sequential line scans for velocity image reconstruction. The ODT signal phase or phase shifts at each pixel can be determined from the complex function, {tilde over (Γ)}ODT(t), which is determined through analytic continuation of the measured interference fringes function, ΓODT(t), by use of a Hilbert transformation, by electronic phase demodulation, by optical means, or a fast Fourier transformation. The phase change in each pixel between axial-line scans is then used to calculate the Doppler frequency shift. Sequential measurements of a single line scan, measurements of sequential line scans or measurements of line scans in sequential frames may be used. Because the time between line scans is much longer than the pixel time window, very small Doppler shifts can be detected with this technique. In addition, spatial resolution and velocity sensitivity are decoupled. Furthermore, because two axial-line scans are compared at the same location, speckle modulations in the fringe signal cancel each other and, therefore, will not affect the phase-difference calculation.

We present a method, based on a single scattering model, to calculate the attenuation coefficient of each pixel in optical coherence tomography (OCT) depth profiles. Numerical simulations were used to determine the model's response to different depths and attenuation coefficients. Experiments were performed on uniform and layered phantoms with varying attenuation coefficients. They were measured by a 1300 nm OCT system and their attenuation coefficients were evaluated by our proposed method and by fitting the OCT slope as the gold standard. Both methods showed largely consistent results for the uniform phantoms. On the layered phantom, only our proposed method accurately estimated the attenuation coefficients. For all phantoms, the proposed method largely reduced the variability of the estimated attenuation coefficients. The method was illustrated on an in-vivo retinal OCT scan, effectively removing common imaging artifacts such as shadowing. By providing localized, per-pixel attenuation coefficients, this method enables tissue characterization based on attenuation coefficient estimates from OCT data. © 2013 Optical Society of America.

In phase-resolved OCT angiography blood flow is detected from phase changes in between A-scans that are obtained from the same location. In ophthalmology, this technique is vulnerable to eye motion. We address this problem by combining inter-B-scan phase-resolved OCT angiography with real-time eye tracking. A tracking scanning laser ophthalmoscope (TSLO) at 840 nm provided eye tracking functionality and was combined with a phase-stabilized optical frequency domain imaging (OFDI) system at 1040 nm. Real-time eye tracking corrected eye drift and prevented discontinuity artifacts from (micro)saccadic eye motion in OCT angiograms. This improved the OCT spot stability on the retina and consequently reduced the phase-noise, thereby enabling the detection of slower blood flows by extending the inter-B-scan time interval. In addition, eye tracking enabled the easy compounding of multiple data sets from the fovea of a healthy volunteer to create high-quality eye motion artifact-free angiograms. High-quality images are presented of two distinct layers of vasculature in the retina and the dense vasculature of the choroid. Additionally we present, for the first time, a phase-resolved OCT angiogram of the mesh-like network of the choriocapillaris containing typical pore openings. © 2012 Optical Society of America.

We present a new method for high-resolution, three-dimensional fluorescence imaging. In contrast to beam-scanning confocal microscopy, where the laser focus must be scanned both laterally and axially to collect a volume, we obtain depth information without the necessity of depth scanning. In this method, the emitted fluorescence is collected in the backward direction and is sent through a phase plate that encodes the depth information into the phase of a spectrally resolved interference pattern. We demonstrate that decoding this phase information allows for depth localization accuracy better than 4 μm over a 500 μm depth-of-field. In a high numerical aperture configuration with a much smaller depth of field, a localization accuracy of tens of nanometers can be achieved. This approach is ideally suited for miniature endoscopes, where space limitations at the endoscope tip render depth scanning difficult. We illustrate the potential for 3D visualization of complex biological samples by constructing a threedimensional volume of the microvasculature of ex vivo murine heart tissue from a single 2D scan. © 2012 Optical Society of America.

Current OCT devices provide three-dimensional (3D) in-vivo images of the human retina. The resulting very large data sets are difficult to manually assess. Automated segmentation is required to automatically process the data and produce images that are clinically useful and easy to interpret. In this paper, we present a method to segment the retinal layers in these images. Instead of using complex heuristics to define each layer, simple features are defined and machine learning classifiers are trained based on manually labeled examples. When applied to new data, these classifiers produce labels for every pixel. After regularization of the 3D labeled volume to produce a surface, this results in consistent, three-dimensionally segmented layers that match known retinal morphology. Six labels were defined, corresponding to the following layers: Vitreous, retinal nerve fiber layer (RNFL), ganglion cell layer & inner plexiform layer, inner nuclear layer & outer plexiform layer, photoreceptors & retinal pigment epithelium and choroid. For both normal and glaucomatous eyes that were imaged with a Spectralis (Heidelberg Engineering) OCT system, the five resulting interfaces were compared between automatic and manual segmentation. RMS errors for the top and bottom of the retina were between 4 and 6 μm, while the errors for intra-retinal interfaces were between 6 and 15 μm. The resulting total retinal thickness maps corresponded with known retinal morphology. RNFL thickness maps were compared to GDx (Carl Zeiss Meditec) thickness maps. Both maps were mostly consistent but local defects were better visualized in OCT-derived thickness maps.

We demonstrate that tissue structure, birefringence, and blood flow can be imaged simultaneously by use of techniques of polarization-sensitive optical coherence tomography and phase-resolved optical Doppler tomography. An efficient data-acquisition procedure is implemented that optimizes the concurrent processing and display of all three image types. Images of in vivo human skin acquired with a high-speed fiber-based system are presented. (C) 2002 Optical Society of America.

We used a novel phase-resolved optical Doppler tomographic (ODT) technique with very high flow-velocity sensitivity (10 mu m/s) and high spatial resolution (10 mu m) to image blood flow in port-wine stain (PWS) birth-marks in human skin. in addition to the regular ODT velocity and structural images, we use the variance of blood flow velocity to map the PWS vessels. Our device combines ODT and therapeutic systems such that PWS blood flow can be monitored in situ before and after laser treatment. To the authors' knowledge this is the first clinical application of ODT to provide a fast semiquantitative evaluation of the efficacy of PWS laser therapy in situ and in real time. (C) 2000 Optical Society of America.