Cone-beam X-ray Phase Contrast Tomography of Biological Samples

Cone-beam X-ray Phase Contrast Tomography of Biological Samples

Author: Matthias Bartels

Publisher: Universitätsverlag Göttingen

Published: 2013

Total Pages: 220

ISBN-13: 3863951344

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Three-dimensional information of entire objects can be obtained by the remarkable technique of computed tomography (CT). In combination with phase sensitive X-ray imaging high contrast for soft tissue structures can be achieved as opposed to CT based on classical radiography. In this work biological samples ranging from micrometer sized single cells over multi-cellular nerve tissue to entire millimeter sized organs are investigated by use of cone-beam propagationbased X-ray phase contrast. Optimization with respect to contrast, resolution and field of view is achieved by addressing instrumentation, sample preparation and phase reconstruction techniques. By using laboratory sources functional soft tissue within the bony capsule of mouse cochleae is visualized in 3D with unprecedented image quality. At synchrotron storage rings the technique reveals more than 1000 axons running in parallel within a mouse nerve and enables doseefficient three-dimensional cellular imaging as well as two-dimensional imaging at high resolutions below 50 nm.


Book Synopsis Cone-beam X-ray Phase Contrast Tomography of Biological Samples by : Matthias Bartels

Download or read book Cone-beam X-ray Phase Contrast Tomography of Biological Samples written by Matthias Bartels and published by Universitätsverlag Göttingen. This book was released on 2013 with total page 220 pages. Available in PDF, EPUB and Kindle. Book excerpt: Three-dimensional information of entire objects can be obtained by the remarkable technique of computed tomography (CT). In combination with phase sensitive X-ray imaging high contrast for soft tissue structures can be achieved as opposed to CT based on classical radiography. In this work biological samples ranging from micrometer sized single cells over multi-cellular nerve tissue to entire millimeter sized organs are investigated by use of cone-beam propagationbased X-ray phase contrast. Optimization with respect to contrast, resolution and field of view is achieved by addressing instrumentation, sample preparation and phase reconstruction techniques. By using laboratory sources functional soft tissue within the bony capsule of mouse cochleae is visualized in 3D with unprecedented image quality. At synchrotron storage rings the technique reveals more than 1000 axons running in parallel within a mouse nerve and enables doseefficient three-dimensional cellular imaging as well as two-dimensional imaging at high resolutions below 50 nm.


Cone-beam x-ray phase-contrast tomography for the observation of single cells in whole organs

Cone-beam x-ray phase-contrast tomography for the observation of single cells in whole organs

Author: Martin Krenkel

Publisher: Göttingen University Press

Published: 2015

Total Pages: 238

ISBN-13: 3863952510

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X-ray imaging enables the nondestructive investigation of interior structures in otherwise opaque samples. In particular the use of computed tomography (CT) allows for arbitrary virtual slices through the object and 3D information about intricate structures can be obtained. However, when it comes to image very small structures like single cells, the classical CT approach is limited by the weak absorption of soft-tissue. The use of phase information, encoded in measureable intensity images by free-space propagation of coherent x-rays, allows a huge increase in contrast, which enables 3D reconstructions at higher resolutions. In this work the application of propagation-based phase-contrast tomography to lung tissue samples is demonstrated in close to in vivo conditions. Reconstructions of the lung structure of whole mice at down to 5 μm resolution are obtained at a selfbuilt CT setup, which is based on a liquid-metal jet x-ray source. To reach even higher resolutions, synchrotron radiation in combination with suitable holographic phase-retrieval algorithms is employed. Due to optimized cone-beam geometry, field of view and resolution can be varied over a wide range of parameters, so that information on different length scales can be achieved, covering several millimeters field of view down to a 3D resolution of 50 nm. Thus, the sub-cellular 3D imaging of single cells embedded in large pieces of tissue is enabled, which paves the way for future biomedical research.


Book Synopsis Cone-beam x-ray phase-contrast tomography for the observation of single cells in whole organs by : Martin Krenkel

Download or read book Cone-beam x-ray phase-contrast tomography for the observation of single cells in whole organs written by Martin Krenkel and published by Göttingen University Press. This book was released on 2015 with total page 238 pages. Available in PDF, EPUB and Kindle. Book excerpt: X-ray imaging enables the nondestructive investigation of interior structures in otherwise opaque samples. In particular the use of computed tomography (CT) allows for arbitrary virtual slices through the object and 3D information about intricate structures can be obtained. However, when it comes to image very small structures like single cells, the classical CT approach is limited by the weak absorption of soft-tissue. The use of phase information, encoded in measureable intensity images by free-space propagation of coherent x-rays, allows a huge increase in contrast, which enables 3D reconstructions at higher resolutions. In this work the application of propagation-based phase-contrast tomography to lung tissue samples is demonstrated in close to in vivo conditions. Reconstructions of the lung structure of whole mice at down to 5 μm resolution are obtained at a selfbuilt CT setup, which is based on a liquid-metal jet x-ray source. To reach even higher resolutions, synchrotron radiation in combination with suitable holographic phase-retrieval algorithms is employed. Due to optimized cone-beam geometry, field of view and resolution can be varied over a wide range of parameters, so that information on different length scales can be achieved, covering several millimeters field of view down to a 3D resolution of 50 nm. Thus, the sub-cellular 3D imaging of single cells embedded in large pieces of tissue is enabled, which paves the way for future biomedical research.


Cone-beam X-ray Phase-contrast Tomography for the Observation of Single Cells in Whole Organs

Cone-beam X-ray Phase-contrast Tomography for the Observation of Single Cells in Whole Organs

Author: Martin Krenkel

Publisher:

Published: 2015

Total Pages:

ISBN-13:

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X-ray imaging enables the nondestructive investigation of interior structures in otherwise opaque samples. In particular the use of computed tomography (CT) allows for arbitrary virtual slices through the object and 3D information about intricate structures can be obtained. However, when it comes to image very small structures like single cells, the classical CT approach is limited by the weak absorption of soft-tissue. The use of phase information, encoded in measureable intensity images by free-space propagation of coherent x-rays, allows a huge increase in contrast, which enables 3D reco ...


Book Synopsis Cone-beam X-ray Phase-contrast Tomography for the Observation of Single Cells in Whole Organs by : Martin Krenkel

Download or read book Cone-beam X-ray Phase-contrast Tomography for the Observation of Single Cells in Whole Organs written by Martin Krenkel and published by . This book was released on 2015 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: X-ray imaging enables the nondestructive investigation of interior structures in otherwise opaque samples. In particular the use of computed tomography (CT) allows for arbitrary virtual slices through the object and 3D information about intricate structures can be obtained. However, when it comes to image very small structures like single cells, the classical CT approach is limited by the weak absorption of soft-tissue. The use of phase information, encoded in measureable intensity images by free-space propagation of coherent x-rays, allows a huge increase in contrast, which enables 3D reco ...


Nanoscale Photonic Imaging

Nanoscale Photonic Imaging

Author: Tim Salditt

Publisher: Springer Nature

Published: 2020-06-09

Total Pages: 634

ISBN-13: 3030344134

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This open access book, edited and authored by a team of world-leading researchers, provides a broad overview of advanced photonic methods for nanoscale visualization, as well as describing a range of fascinating in-depth studies. Introductory chapters cover the most relevant physics and basic methods that young researchers need to master in order to work effectively in the field of nanoscale photonic imaging, from physical first principles, to instrumentation, to mathematical foundations of imaging and data analysis. Subsequent chapters demonstrate how these cutting edge methods are applied to a variety of systems, including complex fluids and biomolecular systems, for visualizing their structure and dynamics, in space and on timescales extending over many orders of magnitude down to the femtosecond range. Progress in nanoscale photonic imaging in Göttingen has been the sum total of more than a decade of work by a wide range of scientists and mathematicians across disciplines, working together in a vibrant collaboration of a kind rarely matched. This volume presents the highlights of their research achievements and serves as a record of the unique and remarkable constellation of contributors, as well as looking ahead at the future prospects in this field. It will serve not only as a useful reference for experienced researchers but also as a valuable point of entry for newcomers.


Book Synopsis Nanoscale Photonic Imaging by : Tim Salditt

Download or read book Nanoscale Photonic Imaging written by Tim Salditt and published by Springer Nature. This book was released on 2020-06-09 with total page 634 pages. Available in PDF, EPUB and Kindle. Book excerpt: This open access book, edited and authored by a team of world-leading researchers, provides a broad overview of advanced photonic methods for nanoscale visualization, as well as describing a range of fascinating in-depth studies. Introductory chapters cover the most relevant physics and basic methods that young researchers need to master in order to work effectively in the field of nanoscale photonic imaging, from physical first principles, to instrumentation, to mathematical foundations of imaging and data analysis. Subsequent chapters demonstrate how these cutting edge methods are applied to a variety of systems, including complex fluids and biomolecular systems, for visualizing their structure and dynamics, in space and on timescales extending over many orders of magnitude down to the femtosecond range. Progress in nanoscale photonic imaging in Göttingen has been the sum total of more than a decade of work by a wide range of scientists and mathematicians across disciplines, working together in a vibrant collaboration of a kind rarely matched. This volume presents the highlights of their research achievements and serves as a record of the unique and remarkable constellation of contributors, as well as looking ahead at the future prospects in this field. It will serve not only as a useful reference for experienced researchers but also as a valuable point of entry for newcomers.


X-Ray Phase-Contrast Tomography

X-Ray Phase-Contrast Tomography

Author: Luca Brombal

Publisher: Springer Nature

Published: 2020-11-16

Total Pages: 140

ISBN-13: 3030604330

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X-ray imaging is a corner stone of breast cancer diagnosis. By exploiting the phase shift of X-rays rather than their attenuation, phase-contrast tomography has the potential to dramatically increase the visibility of small and low contrast features, thus leading to better diagnosis. This thesis presents research on the first synchrotron-based project developing a clinical phase-contrast breast computed tomography (CT) setup at Elettra, the Italian Syncrotron Radiation Facility. This book includes a comprehensive theoretical background on propagation-based phase-contrast imaging, exploring and extending the most recent image formation models. Along with theory, many practical implementation and optimization issues, ranging from detector-specific processing to setup geometry, are tackled on the basis of a large number of experimental evidences. Most of the modelling results and data analysis have general validity, being a valuable framework for optimization of phase-contrast setups. Results obtained at synchrotron are also compared with "real world" laboratory sources: both a first-of-its-kind comparison with one of the few hospital breast CT systems and a state-of-the-art implementation of monochromatic phase-contrast micro-tomography with a conventional rotating anode source are presented. On a more general level, this work sheds a light on the importance of synchrotron-based clinical programs, which are key to trigger the long-anticipated transition of phase-contrast imaging from synchrotrons to hospitals.


Book Synopsis X-Ray Phase-Contrast Tomography by : Luca Brombal

Download or read book X-Ray Phase-Contrast Tomography written by Luca Brombal and published by Springer Nature. This book was released on 2020-11-16 with total page 140 pages. Available in PDF, EPUB and Kindle. Book excerpt: X-ray imaging is a corner stone of breast cancer diagnosis. By exploiting the phase shift of X-rays rather than their attenuation, phase-contrast tomography has the potential to dramatically increase the visibility of small and low contrast features, thus leading to better diagnosis. This thesis presents research on the first synchrotron-based project developing a clinical phase-contrast breast computed tomography (CT) setup at Elettra, the Italian Syncrotron Radiation Facility. This book includes a comprehensive theoretical background on propagation-based phase-contrast imaging, exploring and extending the most recent image formation models. Along with theory, many practical implementation and optimization issues, ranging from detector-specific processing to setup geometry, are tackled on the basis of a large number of experimental evidences. Most of the modelling results and data analysis have general validity, being a valuable framework for optimization of phase-contrast setups. Results obtained at synchrotron are also compared with "real world" laboratory sources: both a first-of-its-kind comparison with one of the few hospital breast CT systems and a state-of-the-art implementation of monochromatic phase-contrast micro-tomography with a conventional rotating anode source are presented. On a more general level, this work sheds a light on the importance of synchrotron-based clinical programs, which are key to trigger the long-anticipated transition of phase-contrast imaging from synchrotrons to hospitals.


Biomedical Imaging

Biomedical Imaging

Author: Tim Salditt

Publisher: Walter de Gruyter GmbH & Co KG

Published: 2017-10-23

Total Pages: 358

ISBN-13: 3110426692

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Covering both physical as well as mathematical and algorithmic foundations, this graduate textbook provides the reader with an introduction into modern biomedical imaging and image processing and reconstruction. These techniques are not only based on advanced instrumentation for image acquisition, but equally on new developments in image processing and reconstruction to extract relevant information from recorded data. To this end, the present book offers a quantitative treatise of radiography, computed tomography, and medical physics. Contents Introduction Digital image processing Essentials of medical x-ray physics Tomography Radiobiology, radiotherapy, and radiation protection Phase contrast radiography Object reconstruction under nonideal conditions


Book Synopsis Biomedical Imaging by : Tim Salditt

Download or read book Biomedical Imaging written by Tim Salditt and published by Walter de Gruyter GmbH & Co KG. This book was released on 2017-10-23 with total page 358 pages. Available in PDF, EPUB and Kindle. Book excerpt: Covering both physical as well as mathematical and algorithmic foundations, this graduate textbook provides the reader with an introduction into modern biomedical imaging and image processing and reconstruction. These techniques are not only based on advanced instrumentation for image acquisition, but equally on new developments in image processing and reconstruction to extract relevant information from recorded data. To this end, the present book offers a quantitative treatise of radiography, computed tomography, and medical physics. Contents Introduction Digital image processing Essentials of medical x-ray physics Tomography Radiobiology, radiotherapy, and radiation protection Phase contrast radiography Object reconstruction under nonideal conditions


Multi-scale 3D Virtual Histology Via Phase-contrast X-ray Tomography with Synchrotron Radiation

Multi-scale 3D Virtual Histology Via Phase-contrast X-ray Tomography with Synchrotron Radiation

Author: Jasper Frohn

Publisher: Universitätsverlag Göttingen

Published: 2023

Total Pages: 148

ISBN-13: 386395601X

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To this day, the standard method for investigating biological tissue with cellular resolution is the examination under a light microscope, first denoted as histology by Karl Meyer in 1819. Despite the enormous success and importance of histology, it has two major disadvantages. Firstly, the specimen must be physically cut into thin sections due to the limited penetrating power of optical light, and secondly, additional staining of the specimen is required to achieve sufficient image contrast. Both disadvantages can be overcome by the non-destructive method of propagation-based X-ray phase-contrast tomography. While the mechanism of phase-contrast provides sufficient image contrast to image single cells, a tomographic imaging scheme with penetrating X-rays allows for an undamaged sample by virtually slicing the reconstructed 3D sample volume. In this work, the holotomography setup of the synchrotron endstation „GINIX“ (The Göttingen Instrument for Nanoscale-Imaging with X-Rays) was extended to a multi-scale X-ray phase-contrast tomography setup suitable for 3D virtual histology by adding two acquisition schemes. Compared to the existing setup, the first additional scheme is a propagation-based microtomography setup, which enlarges the reconstructed 3D volumes by a factor of approx. 64 at a fraction of the acquisition time (ca. 2 min). The second additional scheme aims for higher resolutions. To this end, the X-ray waveguide illumination was combined with photon counting detector with a large field of view and a novel phase reconstruction scheme, which is based on iterative farfield phase retrieval without an „empty-beam correction“ in the detector plane.


Book Synopsis Multi-scale 3D Virtual Histology Via Phase-contrast X-ray Tomography with Synchrotron Radiation by : Jasper Frohn

Download or read book Multi-scale 3D Virtual Histology Via Phase-contrast X-ray Tomography with Synchrotron Radiation written by Jasper Frohn and published by Universitätsverlag Göttingen. This book was released on 2023 with total page 148 pages. Available in PDF, EPUB and Kindle. Book excerpt: To this day, the standard method for investigating biological tissue with cellular resolution is the examination under a light microscope, first denoted as histology by Karl Meyer in 1819. Despite the enormous success and importance of histology, it has two major disadvantages. Firstly, the specimen must be physically cut into thin sections due to the limited penetrating power of optical light, and secondly, additional staining of the specimen is required to achieve sufficient image contrast. Both disadvantages can be overcome by the non-destructive method of propagation-based X-ray phase-contrast tomography. While the mechanism of phase-contrast provides sufficient image contrast to image single cells, a tomographic imaging scheme with penetrating X-rays allows for an undamaged sample by virtually slicing the reconstructed 3D sample volume. In this work, the holotomography setup of the synchrotron endstation „GINIX“ (The Göttingen Instrument for Nanoscale-Imaging with X-Rays) was extended to a multi-scale X-ray phase-contrast tomography setup suitable for 3D virtual histology by adding two acquisition schemes. Compared to the existing setup, the first additional scheme is a propagation-based microtomography setup, which enlarges the reconstructed 3D volumes by a factor of approx. 64 at a fraction of the acquisition time (ca. 2 min). The second additional scheme aims for higher resolutions. To this end, the X-ray waveguide illumination was combined with photon counting detector with a large field of view and a novel phase reconstruction scheme, which is based on iterative farfield phase retrieval without an „empty-beam correction“ in the detector plane.


Developments in X-ray Tomography

Developments in X-ray Tomography

Author:

Publisher:

Published: 2006

Total Pages: 708

ISBN-13:

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Book Synopsis Developments in X-ray Tomography by :

Download or read book Developments in X-ray Tomography written by and published by . This book was released on 2006 with total page 708 pages. Available in PDF, EPUB and Kindle. Book excerpt:


3d virtual histology of neuronal tissue by propagation-based x-ray phase-contrast tomography

3d virtual histology of neuronal tissue by propagation-based x-ray phase-contrast tomography

Author: Mareike Töpperwien

Publisher: Göttingen University Press

Published: 2018

Total Pages: 286

ISBN-13: 3863953649

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Deciphering the three-dimensional (3d) cytoarchitecture of neuronal tissue is an important step towards understanding the connection between tissue function and structure and determining relevant changes in neurodegenerative diseases. The gold standard in pathology is histology, in which the tissue is examined under a light microscope after serial sectioning and subsequent staining. It is an invasive and labor-intensive technique which is prone to artifacts due to the slicing procedure. While it provides excellent results on the 2d slices, the 3d anatomy can only be determined after aligning the individual sections, leading to a non-isotropic resolution within the tissue. X-ray computed tomography (CT) offers a promising alternative due to its potential resolution and large penetration depth which allows for non-invasive imaging of the sample's 3d density distribution. In classical CT, contrast formation is based on absorption of the x-rays as they pass through the sample. However, weakly absorbing samples like soft tissue from the central nervous system give nearly no contrast. By exploiting the much stronger phase shifts for contrast formation, which the sample induces in a (partially) coherent wavefront, it can be substantially increased. During free-space propagation behind the sample, these phase shifts are converted to a measurable intensity image by interference of the disturbed wave fronts. In this thesis, 3d virtual histology is performed by means of propagation-based x-ray phase-contrast tomography on tissue from the central nervous system of humans and mice. A combination of synchrotron-based and laboratory setups is used to visualize the 3d density distribution on varying lengths scales from the whole organ down to single cells. By comparing and optimizing different preparation techniques and phase-retrieval approaches, even sub-cellular resolution can be reached in mm-sized tissue blocks. The development of an automatic cell segmentation workflow provides access to the 3d cellular distribution within the tissue, enabling the quantification of the cellular arrangement and allowing for extensive statistical analysis based on several thousands to millions of cells. This paves the way for biomedical studies aimed at changes in cellular distribution, e.g., in the course of neurodegenerative diseases such as multiple sclerosis, Alzheimer's disease or ischemic stroke.


Book Synopsis 3d virtual histology of neuronal tissue by propagation-based x-ray phase-contrast tomography by : Mareike Töpperwien

Download or read book 3d virtual histology of neuronal tissue by propagation-based x-ray phase-contrast tomography written by Mareike Töpperwien and published by Göttingen University Press. This book was released on 2018 with total page 286 pages. Available in PDF, EPUB and Kindle. Book excerpt: Deciphering the three-dimensional (3d) cytoarchitecture of neuronal tissue is an important step towards understanding the connection between tissue function and structure and determining relevant changes in neurodegenerative diseases. The gold standard in pathology is histology, in which the tissue is examined under a light microscope after serial sectioning and subsequent staining. It is an invasive and labor-intensive technique which is prone to artifacts due to the slicing procedure. While it provides excellent results on the 2d slices, the 3d anatomy can only be determined after aligning the individual sections, leading to a non-isotropic resolution within the tissue. X-ray computed tomography (CT) offers a promising alternative due to its potential resolution and large penetration depth which allows for non-invasive imaging of the sample's 3d density distribution. In classical CT, contrast formation is based on absorption of the x-rays as they pass through the sample. However, weakly absorbing samples like soft tissue from the central nervous system give nearly no contrast. By exploiting the much stronger phase shifts for contrast formation, which the sample induces in a (partially) coherent wavefront, it can be substantially increased. During free-space propagation behind the sample, these phase shifts are converted to a measurable intensity image by interference of the disturbed wave fronts. In this thesis, 3d virtual histology is performed by means of propagation-based x-ray phase-contrast tomography on tissue from the central nervous system of humans and mice. A combination of synchrotron-based and laboratory setups is used to visualize the 3d density distribution on varying lengths scales from the whole organ down to single cells. By comparing and optimizing different preparation techniques and phase-retrieval approaches, even sub-cellular resolution can be reached in mm-sized tissue blocks. The development of an automatic cell segmentation workflow provides access to the 3d cellular distribution within the tissue, enabling the quantification of the cellular arrangement and allowing for extensive statistical analysis based on several thousands to millions of cells. This paves the way for biomedical studies aimed at changes in cellular distribution, e.g., in the course of neurodegenerative diseases such as multiple sclerosis, Alzheimer's disease or ischemic stroke.


X-Ray Near-Field Holography: Beyond Idealized Assumptions of the Probe

X-Ray Near-Field Holography: Beyond Idealized Assumptions of the Probe

Author: Johannes Hagemann

Publisher: Göttingen University Press

Published: 2017

Total Pages: 152

ISBN-13: 3863953320

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All images are flawed, no matter how good your lenses, mirrors etc. are. Especially in the hard X-ray regime it is challenging to manufacture high quality optics due to the weak interaction of multi-keV photons with matter. This is a tremendous challenge for obtaining high resolution quantitative X-ray microscopy images. In recent years lensless phase contrast imaging has become an alternative to classical absorptionbased imaging methods. Without any optics, the image is formed only by the free space propagation of the wave field. The actual image has to be formed posteriori by numerical reconstruction methods. Advanced phasing methods enable the experimentalist to recover a complex valued specimen from a single or a set of intensity measurement. This would be the ideal case - reality teaches us that there are no ideal imaging conditions. Describing, understanding and circumventing these non ideal imaging conditions and their effects on X-ray near-field holographic (NFH) imaging are the leitmotifs for this thesis. In NFH the non ideal conditions manifest themselves in the illuminating wave field or probe. The probe generally does not satisfy the canonical assumptions of fully coherent and monochromatic radiation emitted by a point source. The main results of this thesis are compiled as a collection of publications. An approach is shown to reconstruct the probe of a X-ray nano-focus setup by a series of measurements of the probe at varied Fresnel number. The following chapter presents a study concerning the reconstruction efficiency in terms of resolution for near- and far-field based lensless imaging. In the following, the reconstruction scheme for the probe is extended to incorporate the effects of partial coherence in the near field. This enables the recovery of the modal structure of the probe which yields a full description of its coherence properties. Giving up the assumption of temporal stability due to the stochastic pulses, delivered by X-ray free electron lasers, the reconstruction of probe and specimen must be achieved from a single shot. A suitable scheme for this purpose is proposed in this work.


Book Synopsis X-Ray Near-Field Holography: Beyond Idealized Assumptions of the Probe by : Johannes Hagemann

Download or read book X-Ray Near-Field Holography: Beyond Idealized Assumptions of the Probe written by Johannes Hagemann and published by Göttingen University Press. This book was released on 2017 with total page 152 pages. Available in PDF, EPUB and Kindle. Book excerpt: All images are flawed, no matter how good your lenses, mirrors etc. are. Especially in the hard X-ray regime it is challenging to manufacture high quality optics due to the weak interaction of multi-keV photons with matter. This is a tremendous challenge for obtaining high resolution quantitative X-ray microscopy images. In recent years lensless phase contrast imaging has become an alternative to classical absorptionbased imaging methods. Without any optics, the image is formed only by the free space propagation of the wave field. The actual image has to be formed posteriori by numerical reconstruction methods. Advanced phasing methods enable the experimentalist to recover a complex valued specimen from a single or a set of intensity measurement. This would be the ideal case - reality teaches us that there are no ideal imaging conditions. Describing, understanding and circumventing these non ideal imaging conditions and their effects on X-ray near-field holographic (NFH) imaging are the leitmotifs for this thesis. In NFH the non ideal conditions manifest themselves in the illuminating wave field or probe. The probe generally does not satisfy the canonical assumptions of fully coherent and monochromatic radiation emitted by a point source. The main results of this thesis are compiled as a collection of publications. An approach is shown to reconstruct the probe of a X-ray nano-focus setup by a series of measurements of the probe at varied Fresnel number. The following chapter presents a study concerning the reconstruction efficiency in terms of resolution for near- and far-field based lensless imaging. In the following, the reconstruction scheme for the probe is extended to incorporate the effects of partial coherence in the near field. This enables the recovery of the modal structure of the probe which yields a full description of its coherence properties. Giving up the assumption of temporal stability due to the stochastic pulses, delivered by X-ray free electron lasers, the reconstruction of probe and specimen must be achieved from a single shot. A suitable scheme for this purpose is proposed in this work.