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
Total Pages: 286
Release: 2018
Genre:
ISBN: 3863953649

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.

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:
Total Pages:
Release: 2018
Genre:
ISBN:

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 alignin...

Nanoscale Photonic Imaging

Nanoscale Photonic Imaging
Author: Tim Salditt
Publisher: Springer Nature
Total Pages: 634
Release: 2020-06-09
Genre: Science
ISBN: 3030344134

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.

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
Total Pages: 148
Release: 2023
Genre:
ISBN: 386395601X

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.

X-ray Phase-Contrast Imaging Using Near-Field Speckles

X-ray Phase-Contrast Imaging Using Near-Field Speckles
Author: Marie-Christine Zdora
Publisher: Springer Nature
Total Pages: 337
Release: 2021-02-16
Genre: Science
ISBN: 3030663299

This thesis presents research on novel X-ray imaging methods that improve the study of specimens with small density differences, revealing their inner structure and density distribution. Exploiting the phase shift of X-rays in a material can significantly increase the image contrast compared to conventional absorption imaging. This thesis provides a practical guide to X-ray phase-contrast imaging with a strong focus on X-ray speckle-based imaging, the most recently developed phase-sensitive method. X-ray speckle-based imaging only requires a piece of abrasive paper in addition to the standard X-ray imaging setup. Its simplicity and robustness combined with the compatibility with laboratory X-ray sources, make it an ideal candidate for wide user uptake in a range of fields. An in-depth overview of the state of the art of X-ray speckle-based imaging and its latest developments is given in this thesis. It, furthermore, explores a broad range of applications, from X-ray optics characterisation, to biomedical imaging for 3D virtual histology and geological studies of volcanic rocks, demonstrating is promising potential. Moreover, the speckle-based technique is placed in the context of other phase-sensitive X-ray imaging methods to assist in the choice of a suitable method, hence serving as a guide and reference work for future users.

Neurohistology and Imaging Techniques

Neurohistology and Imaging Techniques
Author: Radek Pelc
Publisher: Humana
Total Pages: 471
Release: 2020-06-25
Genre: Medical
ISBN: 9781071604267

This volume explores major light microscopic imaging modalities that can be used to view nervous tissue, and discusses the steps needed to use each of them, and ways to interpret the data. The chapters in this book cover topics such as atlasing of insect brain; neuroanatomical tracing through fluorochrome expression; fluorescent probes for amyloids; or optical clearing for ultramicroscopy of GFP-expressing tissues. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Authoritative and cutting-edge, Neurohistology and Imaging Techniques is a valuable resource for both expert and novice users of major light microscopic imaging techniques, and those interested in exploring alternate imaging tools.

X-ray waveguide optics

X-ray waveguide optics
Author: Sarah Hoffmann-Urlaub
Publisher: Göttingen University Press
Total Pages: 134
Release: 2017
Genre:
ISBN: 3863953088

Modern x-ray sources and analysis techniques such as lens less imaging combined with phase retrieval algorithms allow for resolving structure sizes in the nanometer range. For this purpose optics have to be employed, ensuring small focal spot dimensions simultaneously with high photon densities. Furthermore, the wave front behind the optics is required to be smooth enabling for high resolution imaging. Combining all these properties, x-ray waveguides are well suited to perform this task, since the intensity distribution behind the guide is restricted in two dimensions serving as a secondary quasi point-source without wave-front aberrations, showing also a high divergence, suitable for resolving fine features. Importantly, the radiation provided by the waveguide reveals a high degree of coherence, required by many imaging techniques. The waveguide itself consists of an air-filled channel embedded in a solid matrix; typical materials are silicon, germanium or quartz. While the entrance area is nano-sized, the channel length is in the millimeter-range, this way posing challenges to fabricate high aspect ratio geometries. Since the functioning of x-ray waveguides is based on the total reflection at small incident angles, the surface roughness of the channel walls must be as low as possible to avoid scattering and hence loss of intensity. To fulfill these demanding conditions, a process scheme involving spin-coating, electron beam lithography, wet development, reactive ion etching and wafer bonding is optimized within this work. To gain deeper insights into the principle of wave guiding finite difference simulations are performed, also opening access for advanced design considerations such as gratings, tapered and curved channels, or beamsplitters, enabling for constructing novel x-ray tools as for example time delay devices or interferometers. Waveguides in all geometries are tested at synchrotron sources, accomplishing new benchmarks in x-ray optical performance. Here, the x-ray beam leaving the channel, propagates out to a pixel array detector in the far-field region. From the recorded data the intensity distribution in the near-field directly behind the waveguide is reconstructed, revealing an outstanding agreement with the simulations and electron micrographs. Since the radiation field of the waveguide is well-characterized and also tunable to meet the requirements of both the measurement setup and the sample, they are suited of a broad field of applications in coherent x-ray imaging.

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
Author: Marina Eckermann
Publisher: Universitätsverlag Göttingen
Total Pages: 276
Release: 2021
Genre:
ISBN: 3863955285

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.

MicroComputed Tomography

MicroComputed Tomography
Author: Stuart R. Stock
Publisher: CRC Press
Total Pages: 366
Release: 2008-12-23
Genre: Technology & Engineering
ISBN: 1420058770

Due to the availability of commercial laboratory systems and the emergence of user facilities at synchrotron radiation sources, studies of microcomputed tomography or microCT have increased exponentially. MicroComputed Technology provides a complete introduction to the technology, describing how to use it effectively and understand its results. The first part of the book focuses on methodology, covering experimental methods, data analysis, and visualization approaches. The second part addresses various microCT applications, including porous solids, microstructural evolution, soft tissue studies, multimode studies, and indirect analyses. The author presents a sufficient amount of fundamental material so that those new to the field can develop a relative understanding of how to design their own microCT studies. One of the first full-length references dedicated to microCT, this book provides an accessible introduction to field, supplemented with application examples and color images.