Elena Reznikova

High-resolution tomographic imaging of a human cerebellum: comparison of absorption and grating-based phase contrast

Human brain tissue belongs to the most impressive and delicate three-dimensional structures in nature. Its outstanding functional importance in the organism implies a strong need for brain imaging modalities. Although magnetic resonance imaging provides deep insights, its spatial resolution is insufficient to study the structure on the level of individual cells. Therefore, our knowledge of brain microstructure currently relies on two-dimensional techniques, optical and electron microscopy, which generally require severe preparation procedures including sectioning and staining. X-ray absorption microtomography yields the necessary spatial resolution, but since the composition of the different types of brain tissue is similar, the images show only marginal contrast. An alternative to absorption could be X-ray phase contrast, which is known for much better discrimination of soft tissues but requires more intricate machinery. In the present communication, we report an evaluation of the recently developed X-ray grating interferometry technique, applied to obtain phase-contrast as well as absorption-contrast synchrotron radiation-based microtomography of human cerebellum. The results are quantitatively compared with synchrotron radiation-based microtomography in optimized absorption-contrast mode. It is demonstrated that grating interferometry allows identifying besides the blood vessels, the stratum moleculare, the stratum granulosum and the white matter. Along the periphery of the stratum granulosum, we have detected microstructures about 40 µm in diameter, which we associate with the Purkinje cells because of their location, size, shape and density. The detection of individual Purkinje cells without the application of any stain or contrast agent is unique in the field of computed tomography and sets new standards in non-destructive three-dimensional imaging.

Inverse geometry for grating-based x-ray phase-contrast imaging

Phase-contrast imaging using conventional polychromatic x-ray sources and grating interferometers has been developed and demonstrated for x-ray energies up to 60 keV. Here, we conduct an analysis of possible grating configurations for this technique and present further geometrical arrangements not considered so far. An inverse interferometer geometry is investigated that offers significant advantages for grating fabrication and for the application of the method in computed tomography (CT) scanners. We derive and measure the interferometer’s angular sensitivity for both the inverse and the conventional configuration as a function of the sample position. Thereby, we show that both arrangements are equally sensitive and that the highest sensitivity is obtained, when the investigated object is close to the interferometer’s phase grating. We also discuss the question whether the sample should be placed in front of or behind the phase grating. For CT applications, we propose an inverse geometry with the sample ...

Recent developments in X-ray Talbot interferometry at ESRF-ID19

In this paper we describe the design of different X-ray Talbot interferometers that have been built at the tomography beamline ID19 of the European Synchrotron Radiation Facility (ESRF) in Grenoble, France, and give a short review of performance characteristics, of current developments, and of the results obtained with these instruments so far. Among the applications so far, soft-tissue imaging has been a particular focus, as demonstrated in a recent paper by Schulz et al. (J. Roy. Soc. Interface, in press).

Planar sets of cross x-ray refractive lenses from SU-8 polymer

Sets of planar SU-8 cross lenses focusing in two directions have been fabricated by tilted deep X-ray lithography using an X-ray mask with tilted absorber structures. The profile of the absorber structures on the mask take into account the lithographic peculiarities of SU-8 resist to reproduce the designed profile of the lens elements exactly. The cross lenses are placed on one substrate and have identical focal distances, which allow to scan the spectral range from 5 keV to 30 keV by stepping the lens substrate from one lens to the next. Another set of cross lenses was developed with different quasi-parabolic profiles to obtain a large focus depth (up to several centimeters) with uniform intensity distribution in the micron focal spot. This together with the stepping possibilities between lenses satisfies the requirement of static spectroscopy experiments. For the truncated parabolic profile, these cross lenses consist of separate segments arranged in a new mosaic form. In comparison with the known “fern”-like kinoform profile, the lenses have been developed with smaller gain loss. The testing of the new sets have been performed at the undulator ID-18F and ID-22 beamlines (ESRF, Grenoble, France) and the experimental results are compared to simulations.

Parabolic crossed planar polymeric x-ray lenses

The principles of design and manufacturing of the polymer planar x-ray lenses focusing in one and two directions, as well as the peculiarities of optical behaviors and the results of the lens test are reported in this paper. The methods of electron and deep x-ray lithography used in lens manufacturing allow the manufacture of ten or more x-ray lenses on one substrate; the lenses show focal lengths down to several centimeters for photon energies between 5 and 40 keV. The measured focus size was 105 nm for a linear lens with an intensity gain of about 407, and 300 x 770 nm for a crossed lens with an intensity gain of 6470.

Imaging of Metastatic Lymph Nodes by X-ray Phase-Contrast Micro-Tomography

Invasive cancer causes a change in density in the affected tissue, which can be visualized by x-ray phase-contrast tomography. However, the diagnostic value of this method has so far not been investigated in detail. Therefore, the purpose of this study was, in a blinded manner, to investigate whether malignancy could be revealed by non-invasive x-ray phase-contrast tomography in lymph nodes from breast cancer patients. Seventeen formalin-fixed paraffin-embedded lymph nodes from 10 female patients (age range 37–83 years) diagnosed with invasive ductal carcinomas were analyzed by X-ray phase-contrast tomography. Ten lymph nodes had metastatic deposits and 7 were benign. The phase-contrast images were analyzed according to standards for conventional CT images looking for characteristics usually only visible by pathological examinations. Histopathology was used as reference. The result of this study was that the diagnostic sensitivity of the image analysis for detecting malignancy was 100% and the specificity was 87%. The positive predictive value was 91% for detecting malignancy and the negative predictive value was 100%. We conclude that x-ray phase-contrast imaging can accurately detect density variations to obtain information regarding lymph node involvement previously inaccessible with standard absorption x-ray imaging.

Submicron focusing of high-energy x-rays with Ni refractive lenses

We report the results on experimental study of optical properties of Ni refractive lenses made by deep X-ray lithography and LIGA techniques. One- and two-dimensional lenses were tested at the ESRF ID15 beamline using wide energy spectrum from 40 keV to 220 keV. The focusing properties in terms of focal length, size of the focal spot/line and gain were studied. Sub micrometer focusing was measured in the energy range from 40 to 150 keV. The measured lens parameters were compared with ray-tracing analysis.

X-ray prism lenses with large apertures

Existing refractive X-ray lenses are characterized by either small apertures due to high absorption in the border areas. They can only be used with synchrotron sources, offering high brilliance. By increasing transparency and aperture the range of applications will expand, common X-ray tubes might turn out to be reasonable X-ray sources in an application with X-ray lenses. A basic concept that meets the demands is an X-ray Fresnel lens. But, Fresnel X-ray lenses are hard to fabricate, since the smaller lens structures need to be produced with extremely high aspect ratios. As an alternative, the Fresnel structures can be replaced by an array of prism-shaped structures. In particular equilateral triangular structures are easier to fabricate and additionally give a higher surface-volume-ratio, increasing transparency. At the Institute for Microstructure Technology the development of such prism lenses is under way. Due to the physical properties of X-rays, several thousands of precisely arranged prisms with large aspect ratio and smooth sidewalls are needed for a single X-ray lens. Therefore, direct X-ray lithography is used to fabricate the SU-8 microstructures. The length of one single prism edge is of the order of 10 μm. One single prismatic X-ray lens consists of up to 60.000 prisms. With the appropriate X-ray mask, refractive X-ray lenses with an aperture of up to 2 mm, for a source distance of 350 mm and a working distance of 350 mm are being produced, assuming a point-shaped source. These X-ray prism lenses are not optimized for smallest focal diameter, but designed to illuminate samples in X-ray optical systems. Most important in this application is an as high transparency as possible.

X-ray Lenses Fabricated by LIGA Technology

X-ray refractive optical lens systems have been successfully elaborated, designed, fabricated at the Institute for Microstructure Technology at the Forschungszentrum Karlsruhe (Germany) using LIGA technology in recent years. The lenses are structured in a SU-8 polymer. The capability of the LIGA technique to create an arbitrary profile of the focusing microstructures allow the fabrication of lenses with different curvature radius of parabolic geometry, minimized absorption and a large depth of focus. Also a set of planar lens systems on one substrate can be realized with 17 lenses providing identical focal distances for different X-ray energies from 2 to over 100 keV. Nickel lenses fabricated by electroforming using polymer templates can be applied for energies larger than 80 keV. The parabolic crossed lenses are used for 2D nano focusing of monochromatic beams. The quasi-parabolic crossed lenses with a submicron focus and a focus depth of the centimetre range can be used as an achromatic system. Mosaic truncated parabolic lenses with a focusing aperture up to 1 mm are made to increase the X-ray intensity in the focused spot.

Near-diffraction limited coherent x-ray focusing using planar refractive lenses made of epoxy SU-8 resist

We present results on comprehensive studies of high resolution SU-8 planar refractive lenses. Lens optical properties were investigated using coherent high energy X-ray radiation. Resolution of about 270 nm was measured for the lens consisting of 31 individual lenses at energy 14 keV. Coherent properties of the set-up permit to resolve near-focus fine structure, which is determined by tiny aberrations caused by lens imperfections close to the parabola apex. This study allows understanding as far SR deep lithography as possible can maintaine to close tolerances for lens parameters. Two-dimensional focusing crossed lenses were tested and imaging experiments in projection and imaging mode were conducted. Radiation stability test was performed and conclusions on the applicability of SU-8 lenses were done.