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 PDF

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.

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 PDF

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.

Multiscale X-ray Structural Analysis of Cardiac Cells and Tissues

Multiscale X-ray Structural Analysis of Cardiac Cells and Tissues PDF

Author: Marius Reichardt

Publisher: Universitätsverlag Göttingen

Published: 2022

Total Pages: 234

ISBN-13: 3863955366

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The cardiac function relies on an intricate molecular and cellular three-dimensional (3d) architecture of a complex, dense and co-dependent cellular network. Structural alterations of the cardiac structure can affect its essential function and lead to severe dysfunction of the organ. Cardiovascular diseases are the main cause of death worldwide with a rising incidence. However, it is not possible to give a generalized answer how the heart is formed. Up to now, cardiac structure as well as physiologic and disease-related tissue alterations of the tissue are mainly investigated by established 2d imaging methods such as optical microscopy or electron microscopy. This work presents a multiscale and multimodal X-ray imaging approach, which allows to probe the heart structure from the scale of entire intact murine hearts to the molecular organisation of the sarcomer structure. While the molecular structure of the actomyosin complex is probed by scanning X-ray diffraction, the 3d arrangement of the cellular network is investigated by propagation-based X-ray phase-contrast tomography. In this context, the concept of 3d virtual histology of cardiac tissue by X-ray phase-contrast tomography using laboratory sources as well as highly coherent synchrotron radiation is being further developed.

Advancing the Characterization of Neuronal Cyto-Architecture by X-ray Phase-Contrast Tomography

Advancing the Characterization of Neuronal Cyto-Architecture by X-ray Phase-Contrast Tomography PDF

Author: Marina Eckermann

Publisher: Universitätsverlag Göttingen

Published: 2021

Total Pages: 276

ISBN-13: 3863955285

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To bring physiology and pathology of the human brain into better micro-anatomical and histological context, studies with different methodologies are required. Established techniques such as electron microscopy or histology show limitations in view of invasiveness, labor-intense and artifact-prone sample preparation, as well as an adequate ratio between resolution and volume throughput. For this reason, X-ray phase-contrast tomography (PC-CT) has been proposed as a three-dimensional non-destructive imaging technique, which requires less effort in sample preparation and can assess larger volumes. Furthermore, it offers quantitative electron density based contrast even for unstained tissue. Up to now, however, PC-CT studies fell short in number of samples, so that structural alterations caused by neurodegenerative diseases cannot be distinguished from physiological inter-subject variations. In this thesis, the scalability of PC-CT with respect to the required number of samples and resolution-to-volume-throughput is demonstrated, and the methodology is advanced with respect to data acquisition, processing and segmentation. In addition to the human cerebellum, cortex and hippocampus are studied. Concerning quantification and analysis of PC-CT data, this work introduces optimal transport analysis to obtain quantitative metrics of the cyto-architecture and to identify changes due to neurodegenerative diseases. For the case of Alzheimer’s disease, this workflow reveals a yet undescribed compactification of granular cells in the human hippocampus. This thesis also provides optimized configurations to study neural tissues with laboratory instrumentation, and – finally – provides new correlative imaging approaches, in particular with scanning electron microscopy.

Vertebrate Skeletal Histology and Paleohistology

Vertebrate Skeletal Histology and Paleohistology PDF

Author: Vivian de Buffrénil

Publisher: CRC Press

Published: 2021-06-24

Total Pages: 838

ISBN-13: 1351189581

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Vertebrate Skeletal Histology and Paleohistology summarizes decades of research into the biology and biological meaning of hard tissues, in both living and extinct vertebrates. In addition to outlining anatomical diversity, it provides fundamental phylogenetic and evolutionary contexts for interpretation. An international team of leading authorities review the impact of ontogeny, mechanics, and environment in relation to bone and dental tissues. Synthesizing current advances in the biological problems of growth, metabolism, evolution, ecology, and behavior, this comprehensive and authoritative volume is built upon a foundation of concepts and technology generated over the past fifty years.

Nanoscale Photonic Imaging

Nanoscale Photonic Imaging PDF

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.

Coherent X-Ray Optics

Coherent X-Ray Optics PDF

Author: David Paganin

Publisher: OUP Oxford

Published: 2006-01-12

Total Pages: 424

ISBN-13: 019152431X

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This book gives a thorough treatment of the rapidly-expanding field of coherent x-ray optics, which has recently experienced something of a renaissance with the availability of third-generation synchrotron sources. It is the first book of its kind. The author begins with a treatment of the fundamentals of x-ray diffraction for both coherent and partially coherent radiation, together with the interactions of x-rays with matter. X-ray sources, optics elements and detectors are then discussed, with an emphasis on their role in coherent x-ray optics. Various facets of coherent x-ray imaging are then discussed, including holography, interferometry, self imaging, phase contrast and phase retrieval. Lastly, the foundations of the new field of singular x-ray optics are examined. Most topics are developed from first principles, with numerous references given to the contemporary research literature. This book will be useful to x-ray physicists and students, together with optical physicists and engineers who wish to learn more about the fascinating subject of coherent x-ray optics.

Computed Tomography

Computed Tomography PDF

Author: Willi A. Kalender

Publisher: John Wiley & Sons

Published: 2011-07-07

Total Pages: 372

ISBN-13: 3895786446

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The book offers a comprehensive and user-oriented description of the theoretical and technical system fundamentals of computed tomography (CT) for a wide readership, from conventional single-slice acquisitions to volume acquisition with multi-slice and cone-beam spiral CT. It covers in detail all characteristic parameters relevant for image quality and all performance features significant for clinical application. Readers will thus be informed how to use a CT system to an optimum depending on the different diagnostic requirements. This includes a detailed discussion about the dose required and about dose measurements as well as how to reduce dose in CT. All considerations pay special attention to spiral CT and to new developments towards advanced multi-slice and cone-beam CT. For the third edition most of the contents have been updated and latest topics like dual source CT, dual energy CT, flat detector CT and interventional CT have been added. The enclosed CD-ROM again offers copies of all figures in the book and attractive case studies, including many examples from the most recent 64-slice acquisitions, and interactive exercises for image viewing and manipulation. This book is intended for all those who work daily, regularly or even only occasionally with CT: physicians, radiographers, engineers, technicians and physicists. A glossary describes all the important technical terms in alphabetical order. The enclosed DVD again offers attractive case studies, including many examples from the most recent 64-slice acquisitions, and interactive exercises for image viewing and manipulation. This book is intended for all those who work daily, regularly or even only occasionally with CT: physicians, radiographers, engineers, technicians and physicists. A glossary describes all the important technical terms in alphabetical order.

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