A four-grating interferometer for x-ray multi-contrast imaging.

BACKGROUND: X-ray multi-contrast imaging with gratings provides a practical method to detect differential phase and dark-field contrast images in addition to the x-ray absorption image traditionally obtained in laboratory or hospital environments. Systems have been developed for preclinical applications in areas including breast imaging, lung imaging, rheumatoid arthritis hand imaging and kidney stone imaging. PURPOSE: Prevailing x-ray interferometers for multi-contrast imaging include Talbot-Lau interferometers and universal moiré effect-based phase-grating interferometers. Talbot-Lau interferometers suffer from conflict between high interferometer sensitivity and large field of view (FOV) of the object being imaged. A small period analyzer grating is necessary to simultaneously achieve high sensitivity and large FOV within a compact imaging system but is technically challenging to produce for high x-ray energies. Phase-grating interferometers suffer from an intrinsic fringe period ranging from a few micrometers to several hundred micrometers that can hardly be resolved by large area flat panel x-ray detectors. The purpose of this work is to introduce a four-grating x-ray interferometer that simultaneously allows high sensitivity and large FOV, without the need for a small period analyzer grating. METHODS: The four-grating interferometer consists of a source grating placed downstream of and close to the x-ray source, a pair of phase gratings separated by a fixed distance placed downstream of the source grating, and an analyzer grating placed upstream of and close to the x-ray detector. The object to be imaged is placed upstream of and close to the phase-grating pair. The distance between the source grating and the phase-grating pair is designed to be far larger than that between the phase-grating pair and the analyzer grating to promote simultaneously high sensitivity and large FOV. The method was evaluated by constructing a four-grating interferometer with an 8 µm period source grating, a pair of phase gratings of 2.4 µm period, and an 8 µm period analyzer grating. RESULTS: The fringe visibility of the four-grating interferometer was measured to be ≈24% at 40 kV and ≈18% at 50 kV x-ray tube operating voltage. A quartz bead of 6 mm diameter was imaged to compare the theoretical and experimental phase contrast signal with good agreement. Kidney stone specimens were imaged to demonstrate the potential of such a system for classification of kidney stones. CONCLUSIONS: The proposed four-grating interferometer geometry enables a compact x-ray multi-contrast imaging system with simultaneously high sensitivity and large FOV. Relaxation of the requirement for a small period analyzer grating makes it particularly suitable for high x-ray energy applications such as abdomen and chest imaging.

The role of nonmuscle myosin 2A and 2B in the regulation of mesenchymal cell contact guidance.

Contact guidance refers to the ability of cells to sense the geometrical features of the microenvironment and respond by changing their shape and adopting the appropriate orientation. Inhibition and ablation of nonmuscle myosin 2 (NM2) paralogues have demonstrated their importance for contact guidance. However, the specific roles of the NM2 paralogues have not been systematically studied. In this work we use micropatterned substrates to examine the roles of NM2A and NM2B and to elucidate the relationship of the microenvironment, actomyosin, and microtubules in contact guidance. We show that contact guidance is preserved following loss of NM2B and that expression of NM2A alone is sufficient to establish an appropriate orientation of the cells. Loss of NM2B and overexpression of NM2A result in a prominent cell polarization that is found to be linked to the increased alignment of microtubules with the actomyosin scaffold. Suppression of actomyosin with blebbistatin reduces cell polarity on a flat surface, but not on a surface with contact guidance cues. This indicates that the lost microtubule-actomyosin interactions are compensated for by microtubule-microenvironment interactions, which are sufficient to establish cell polarity through contact guidance.

Correlative Detection of Isolated Single and Multi-Cellular Calcifications in the Internal Elastic Lamina of Human Coronary Artery Samples.

Histopathology protocols often require sectioning and processing of numerous microscopy slides to survey a sample. Trade-offs between workload and sampling density means that small features can be missed. Aiming to reduce the workload of routine histology protocols and the concern over missed pathology in skipped sections, we developed a prototype x-ray tomographic scanner dedicated to rapid scouting and identification of regions of interest in pathology specimens, thereby allowing targeted histopathology analysis to replace blanket searches. In coronary artery samples of a deceased HIV patient, the scanner, called Tomopath, obtained depth-resolved cross-sectional images at 15 µm resolution in a 15-minute scan, which guided the subsequent histological sectioning and microscopy. When compared to a commercial tabletop micro-CT scanner, the prototype provided several-fold contrast-to-noise ratio in 1/11th the scan time. Correlated tomographic and histological images revealed two types of micro calcifications: scattered loose calcifications typically found in atherosclerotic lesions; isolated focal calcifications in one or several cells in the internal elastic lamina and occasionally in the tunica media, which we speculate were the initiation of medial calcification linked to kidney disease, but rarely detected at this early stage due to their similarity to particle contaminants introduced during histological processing, if not for the evidence from the tomography scan prior to sectioning. Thus, in addition to its utility as a scouting tool, in this study it provided complementary information to histological microscopy. Overall, the prototype scanner represents a step toward a dedicated scouting and complementary imaging tool for routine use in pathology labs.

Cryogenic Etching of High Aspect Ratio 400 nm Pitch Silicon Gratings.

The cryogenic process and Bosch process are two widely used processes for reactive ion etching of high aspect ratio silicon structures. This paper focuses on the cryogenic deep etching of 400 nm pitch silicon gratings with various etching mask materials including polymer, Cr, SiO2 and Cr-on-polymer. The undercut is found to be the key factor limiting the achievable aspect ratio for the direct hard masks of Cr and SiO2, while the etch selectivity responds to the limitation of the polymer mask. The Cr-on-polymer mask provides the same high selectivity as Cr and reduces the excessive undercut introduced by direct hard masks. By optimizing the etching parameters, we etched a 400 nm pitch grating to ≈ 10.6 µm depth, corresponding to an aspect ratio of ≈ 53.

A Universal Moiré Effect and Application in X-Ray Phase-Contrast Imaging.

A moiré pattern is created by superimposing two black-and-white or gray-scale patterns of regular geometry, such as two sets of evenly spaced lines. We observed an analogous effect between two transparent phase masks in a light beam which occurs at a distance. This phase moiré effect and the classic moiré effect are shown to be the two ends of a continuous spectrum. The phase moiré effect allows the detection of sub-resolution intensity or phase patterns with a transparent screen. When applied to x-ray imaging, it enables a polychromatic far-field interferometer (PFI) without absorption gratings. X-ray interferometry can non-invasively detect refractive index variations inside an object1-10. Current bench-top interferometers operate in the near field with limitations in sensitivity and x-ray dose efficiency2, 5, 7-10. The universal moiré effect helps overcome these limitations and obviates the need to make hard x-ray absorption gratings of sub-micron periods.

Enhancing Tabletop X-Ray Phase Contrast Imaging with Nano-Fabrication.

X-ray phase-contrast imaging is a promising approach for improving soft-tissue contrast and lowering radiation dose in biomedical applications. While current tabletop imaging systems adapt to common x-ray tubes and large-area detectors by employing absorptive elements such as absorption gratings or monolithic crystals to filter the beam, we developed nanometric phase gratings which enable tabletop x-ray far-field interferometry with only phase-shifting elements, leading to a substantial enhancement in the performance of phase contrast imaging. In a general sense the method transfers the demands on the spatial coherence of the x-ray source and the detector resolution to the feature size of x-ray phase masks. We demonstrate its capabilities in hard x-ray imaging experiments at a fraction of clinical dose levels and present comparisons with the existing Talbot-Lau interferometer and with conventional digital radiography.

Motionless electromagnetic phase stepping versus mechanical phase stepping in x-ray phase-contrast imaging with a compact source.

X-ray phase contrast imaging based on grating interferometers detects the refractive index distribution of an object without relying on radiation attenuation, thereby having the potential for reduced radiation absorption. These techniques belong to the broader category of optical wavefront measurement, which requires stepping the phase of the interference pattern to obtain a pixel-wise map of the phase distortion of the wavefront. While phase stepping traditionally involves mechanical scanning of a grating or mirror, we developed electromagnetic phase stepping (EPS) for imaging with compact sources to obviate the need for mechanical movement. In EPS a solenoid coil is placed outside the x-ray tube to shift its focal spot with a magnetic field, causing a relative movement between the projection of the sample and the interference pattern in the image. Here we present two embodiments of this method. We verified experimentally that electromagnetic and mechanical phase stepping give the same results and attain the same signal-to-noise ratios under the same radiation dose. We found that the relative changes of interference fringe visibility were within 3.0% when the x-ray focal spot was shifted by up to 1.0 mm in either direction. We conclude that when using x-ray tube sources, EPS is an effective means of phase stepping without the need for mechanical movement.

Electrodeposition of Gold to Conformally Fill High Aspect Ratio Nanometric Silicon Grating Trenches: A Comparison of Pulsed and Direct Current Protocols.

Filling high-aspect-ratio trenches with gold is a frequent requirement in the fabrication of x-ray optics as well as micro-electronic components and other fabrication processes. Conformal electrodeposition of gold in sub-micron-width silicon trenches with an aspect ratio greater than 35 over a grating area of several square centimeters is challenging and has not been described in the literature previously. A comparison of pulsed plating and constant current plating led to a gold electroplating protocol that reliably filled trenches for such structures.

Fabrication of 200 nm period hard X-ray phase gratings.

Far field X-ray grating interferometry achieves extraordinary phase sensitivity in imaging weakly absorbing samples, provided that the grating period is within the transverse coherence length of the X-ray source. Here we describe a cost-efficient process to fabricate large area, 100 nm half-pitch hard X-ray phase gratings with an aspect ratio of 32. The nanometric gratings are suitable for ordinary compact X-ray sources having low spatial coherence, as demonstrated by X-ray diffraction experiments.

Motionless phase stepping in X-ray phase contrast imaging with a compact source.

X-ray phase contrast imaging offers a way to visualize the internal structures of an object without the need to deposit significant radiation, and thereby alleviate the main concern in X-ray diagnostic imaging procedures today. Grating-based differential phase contrast imaging techniques are compatible with compact X-ray sources, which is a key requirement for the majority of clinical X-ray modalities. However, these methods are substantially limited by the need for mechanical phase stepping. We describe an electromagnetic phase-stepping method that eliminates mechanical motion, thus removing the constraints in speed, accuracy, and flexibility. The method is broadly applicable to both projection and tomography imaging modes. The transition from mechanical to electromagnetic scanning should greatly facilitate the translation of X-ray phase contrast techniques into mainstream applications.

Flexible retrospective phase stepping in x-ray scatter correction and phase contrast imaging using structured illumination.

The development of phase contrast methods for diagnostic x-ray imaging is inspired by the potential of seeing the internal structures of the human body without the need to deposit any harmful radiation. An efficient class of x-ray phase contrast imaging and scatter correction methods share the idea of using structured illumination in the form of a periodic fringe pattern created with gratings or grids. They measure the scatter and distortion of the x-ray wavefront through the attenuation and deformation of the fringe pattern via a phase stepping process. Phase stepping describes image acquisition at regular phase intervals by shifting a grating in uniform steps. However, in practical conditions the actual phase intervals can vary from step to step and also spatially. Particularly with the advent of electromagnetic phase stepping without physical movement of a grating, the phase intervals are dependent upon the focal plane of interest. We describe a demodulation algorithm for phase stepping at arbitrary and position-dependent (APD) phase intervals without assuming a priori knowledge of the phase steps. The algorithm retrospectively determines the spatial distribution of the phase intervals by a Fourier transform method. With this ability, grating-based x-ray imaging becomes more adaptable and robust for broader applications.

Interferometric hard x-ray phase contrast imaging at 204 nm grating period.

We report on hard x-ray phase contrast imaging experiments using a grating interferometer of approximately 1/10th the grating period achieved in previous studies. We designed the gratings as a staircase array of multilayer stacks which are fabricated in a single thin film deposition process. We performed the experiments at 19 keV x-ray energy and 0.8 µm pixel resolution. The small grating period resulted in clear separation of different diffraction orders and multiple images on the detector. A slitted beam was used to remove overlap of the images from the different diffraction orders. The phase contrast images showed detailed features as small as 10 µm, and demonstrated the feasibility of high resolution x-ray phase contrast imaging with nanometer scale gratings.

Probing coherence in microcavity frequency combs via optical pulse shaping.

Recent investigations of microcavity frequency combs based on cascaded four-wave mixing have revealed a link between the evolution of the optical spectrum and the observed temporal coherence. Here we study a silicon nitride microresonator for which the initial four-wave mixing sidebands are spaced by multiple free spectral ranges (FSRs) from the pump. Additional lines then fill in to yield a comb with single FSR spacing, resulting in partial coherence. By using a pulse shaper to select and manipulate the phase of various subsets of spectral lines, we are able to probe the structure of the coherence within the partially coherent comb. Our data demonstrate strong variation in the degree of mutual coherence between different groups of lines and provide support for a simple model of partially coherent comb formation.

Wide cantilever stiffness range cavity optomechanical sensors for atomic force microscopy.

We report on progress in developing compact sensors for atomic force microscopy (AFM), in which the mechanical transducer is integrated with near-field optical readout on a single chip. The motion of a nanoscale, doubly clamped cantilever was transduced by an adjacent high quality factor silicon microdisk cavity. In particular, we show that displacement sensitivity on the order of 1 fm/(Hz)(1/2) can be achieved while the cantilever stiffness is varied over four orders of magnitude (≈0.01 N/m to ≈290 N/m). The ability to transduce both very soft and very stiff cantilevers extends the domain of applicability of this technique, potentially ranging from interrogation of microbiological samples (soft cantilevers) to imaging with high resolution (stiff cantilevers). Along with mechanical frequencies (> 250 kHz) that are much higher than those used in conventional AFM probes of similar stiffness, these results suggest that our cavity optomechanical sensors may have application in a wide variety of high-bandwidth AFM measurements.

Observation of correlation between route to formation, coherence, noise, and communication performance of Kerr combs.

Microresonator optical frequency combs based on cascaded four-wave mixing are potentially attractive as a multi-wavelength source for on-chip optical communications. In this paper we compare time domain coherence, radio-frequency (RF) intensity noise, and individual line optical communications performance for combs generated from two different silicon nitride microresonators. The comb generated by one microresonator forms directly with lines spaced by a single free spectral range (FSR) and exhibits high coherence, low noise, and excellent 10 Gbit/s optical communications results. The comb generated by the second microresonator forms initially with multiple FSR line spacing, with additional lines later filling to reach single FSR spacing. This comb exhibits degraded coherence, increased intensity noise, and severely degraded communications performance. This study is to our knowledge the first to simultaneously investigate and observe a correlation between the route to comb formation, the coherence, noise, and optical communications performance of a Kerr comb.

Optomechanical transduction of an integrated silicon cantilever probe using a microdisk resonator.

Sensitive transduction of the motion of a microscale cantilever is central to many applications in mass, force, magnetic resonance, and displacement sensing. Reducing cantilever size to nanoscale dimensions can improve the bandwidth and sensitivity of techniques like atomic force microscopy, but current optical transduction methods suffer when the cantilever is small compared to the achievable spot size. Here, we demonstrate sensitive optical transduction in a monolithic cavity-optomechanical system in which a subpicogram silicon cantilever with a sharp probe tip is separated from a microdisk optical resonator by a nanoscale gap. High quality factor (Q ≈ 10(5)) microdisk optical modes transduce the cantilever's megahertz frequency thermally driven vibrations with a displacement sensitivity of ≈4.4 × 10(-16) m/(Hz)(1/2) and bandwidth >1 GHz, and a dynamic range >10(6) is estimated for a 1 s measurement. Optically induced stiffening due to the strong optomechanical interaction is observed, and engineering of probe dynamics through cantilever design and electrostatic actuation is illustrated.

Polarization mode dispersion spectrum measurement via high-speed wavelength-parallel polarimetry.

We report experiments in which wavelength-parallel spectral polarimetry technology is used for measurement of the frequency-dependent polarization mode dispersion (PMD) vector. Experiments have been performed using either a grating spectral disperser, configured to provide 13.6 GHz spectral resolution over a 14 nm optical bandwidth, or a virtually imaged phased array spectral disperser, configured for 1.6 GHz spectral resolution over a 200 GHz band. Our results indicate that the spectral polarimetry data obtained via this approach are of sufficient quality to permit accurate extraction of the PMD spectrum. The wavelength-parallel spectral polarimetry approach allows data acquisition within a few milliseconds.

Optical arbitrary waveform characterization via dual-quadrature spectral shearing interferometry.

We demonstrate a new dual-quadrature spectral shearing interferometry technique appropriate for spectral phase characterization of arbitrary optical waveforms generated by line-by-line shaping of high-repetition- rate (approximately 10 GHz) optical frequency combs. Spectral shearing interferograms are generated through sum-frequency mixing of the frequency comb field with a pair of reference tones generated via intensity modulation of a continuous-wave laser. Although related to the well known SPIDER method, our approach relaxes spectral resolution requirements and operates in a collinear interaction geometry compatible with the use of high sensitivity, aperiodically poled lithium niobate nonlinear waveguide devices.

Direct spectral phase retrieval of ultrashort pulses by double modified one-dimensional autocorrelation traces.

We propose and experimentally demonstrate an original method to analytically retrieve complete spectral phase of ultrashort pulses by measuring two modified interferometric field autocorrelation traces using thick nonlinear crystals with slightly different central phase-matching wavelengths. This new scheme requires no spectrometer, detector array, nor iterative data inversion, and is compatible with periodically poled lithium niobate (PPLN) waveguide technology offering potential for high measurement sensitivity.

Ultrasensitive nonlinear measurements of femtosecond pulses in the telecommunications band by aperiodically poled LiNbO3 waveguides.

We have used aperiodically poled lithium niobate waveguides to perform intensity autocorrelation and frequency-resolved optical gating (FROG) measurements for ultraweak femtosecond pulses at 1.5 microm wavelength. The required pulse energies for intensity autocorrelation and FROG are as low as 52 aJ and 124 aJ, respectively. The corresponding sensitivities are 3.2 x 10(-7) mW(2) and 2.7 x 10(-6) mW(2), about 3-5 orders of magnitude better than the previous records. The high nonlinear conversion efficiency is attributed to the long waveguide structure, and the needed broad phase-matching bandwidth is realized by chirping the poling period. We discuss the theory of intensity autocorrelation and FROG measurements in the presence of different phase-matching bandwidths, and we show, for the first time to our knowledge, that the distorted intensity autocorrelation trace due to a delta-like phase-matching spectrum is described by a modified field autocorrelation function. We also report new experimental results comparing autocorrelation traces measured with chirped and unchirped waveguide samples and demonstrating high-quality FROG measurements for cubic phase waveforms generated in a programmable pulse shaper.