ABSTRACT
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ABSTRACT
We present an approach towards four dimensional (4d) movies of materials, showing dynamic processes within the entire 3d structure. The method is based on tomographic reconstruction on dynamically curved paths using a motion model estimated by optical flow techniques, considerably reducing the typical motion artefacts of dynamic tomography. At the same time we exploit x-ray phase contrast based on free propagation to enhance the signal from micron scale structure recorded with illumination times down to a millisecond (ms). The concept is demonstrated by observing the burning process of a match stick in 4d, using high speed synchrotron phase contrast x-ray tomography recordings. The resulting movies reveal the structural changes of the wood cells during the combustion.
ABSTRACT
In this work, we propose a novel computed tomography (CT) approach for three-dimensional (3D) object reconstruction, based on a generalized tomographic geometry with two-dimensional angular sampling (two angular degrees of freedom). The reconstruction is based on the 3D Radon transform and is compatible with anisotropic beam conditions. This allows isotropic 3D imaging with a source, which can be extended along one direction for increased flux, while high resolution is achieved by a small source size only in the orthogonal direction. This novel scheme for analytical CT is demonstrated by numerical simulations and proof-of-concept experiments. In this way high resolution and coherence along a single direction determines the reconstruction quality of the entire 3D data set, opening up, for example, new opportunities to achieve nanoscale resolution and/or phase contrast with low brilliance sources such as laboratory x-ray or neutron sources.
ABSTRACT
We have used X-ray phase contrast tomography to resolve the structure of uncut, entire myelinated optic, saphenous and sciatic mouse nerves. Intrinsic electron density contrast suffices to identify axonal structures. Specific myelin labeling by an osmium tetroxide stain enables distinction between axon and surrounding myelin sheath. Utilization of spherical wave illumination enables zooming capabilities which enable imaging of entire sciatic internodes as well as identification of sub-structures such as nodes of Ranvier and Schmidt-Lanterman incisures.
Subject(s)
Optic Nerve/ultrastructure , Saphenous Vein/innervation , Saphenous Vein/ultrastructure , Sciatic Nerve/ultrastructure , Animals , Axons/physiology , Imaging, Three-Dimensional , Mice , Mice, Inbred C57BL , Microscopy, Phase-Contrast , Myelin Sheath/physiology , Optic Nerve/anatomy & histology , Osmium Tetroxide/pharmacology , Saphenous Vein/anatomy & histology , Schwann Cells/cytology , Sciatic Nerve/anatomy & histology , Staining and Labeling , Tomography, X-Ray ComputedABSTRACT
Quantitative waveguide-based X-ray phase contrast imaging has been carried out on the level of single, unstained, unsliced and freeze-dried bacterial cells of Bacillus thuringiensis and Bacillus subtilis using hard X-rays of 7.9â keV photon energy. The cells have been prepared in the metabolically dormant state of an endospore. The quantitative phase maps obtained by iterative phase retrieval using a modified hybrid input-output algorithm allow for mass and mass density determinations on the level of single individual endospores but include also large field of view investigations. Additionally, a direct reconstruction based on the contrast transfer function is investigated, and the two approaches are compared. Depending on the field of view and method, a resolution down to 65â nm was achieved at a maximum applied dose of below 5 × 105â Gy. Masses in the range of about â¼110-190â (20)â fg for isolated endospores have been obtained.
ABSTRACT
We demonstrate nanoscale x-ray holographic imaging using optimized illumination wave fronts emitted by x-ray waveguide channels. Mode filtering minimizes wave-front distortions and artifacts encountered in most hard x-ray focusing schemes, enabling quantitative reconstruction of the projected density, as evidenced by a test pattern imaged with a field of view of about 20×40 µm and at 22 nm resolution. The dose efficiency and contrast sensitivity make the optical scheme compatible with samples of intrinsically low contrast, typical for hydrated soft matter. This is demonstrated by imaging bacteria in the hydrated and living state, with quantitative phase contrast revealing dense structures of the bacterial nucleoids associated with compactified DNA. In response to continued irradiation, characteristic changes in these dense structures are observed.
Subject(s)
Deinococcus/cytology , Holography/methods , DNA, Bacterial/chemistry , Holography/instrumentation , Image Processing, Computer-Assisted/methods , Solutions , X-RaysABSTRACT
We have implemented a deterministic method for solving the phase problem in hard x-ray in-line holography which overcomes the twin image problem. The phase distribution in the detector plane is retrieved by using two images with slightly different Fresnel numbers. We then use measured intensities and reconstructed phases in the detection plane to compute the exit wave in the sample plane. No further a priori information like a limited support or the assumption of pure phase objects is necessary so that it can be used for a wide range of complex samples. Using a nano-focused hard x-ray beam half period resolutions better than 30 nm are achieved.
Subject(s)
Holography/instrumentation , Imaging, Three-Dimensional/instrumentation , Radiographic Image Interpretation, Computer-Assisted/instrumentation , Radiography/instrumentation , Subtraction Technique/instrumentation , Equipment Design , Equipment Failure AnalysisABSTRACT
We have analyzed the model organism Caenorhabditis elegans with the help of phase-contrast x-ray tomography. This work combines techniques from x-ray imaging studies of single biological cells by in-line holography with three-dimensional reconstruction and furthermore extends these studies to the multicellular level. To preserve the sub-cellular ultrastructure of the nematodes, we used the near-native sample preparation of high-pressure freezing as commonly used in the field of electron microscopy. For the presented samples, a standard, non-magnifying parallel-beam setting, as well as a magnifying, divergent-beam setting using nanofocusing optics is evaluated based on their tomographic reconstruction potential. In this paper, we address difficulties in sample preparation and issues of image processing. By experimental refinement and through optimized reconstruction procedures, we were able to perform x-ray imaging studies on a living specimen.