Coherent X-ray diffractive imaging on the single-cell-level of microbial samples

Coherent X-ray diffractive imaging on the single-cell-level of microbial samples PDF Author: Robin Niklas Wilke
Publisher: Göttingen University Press
ISBN: 3863951905
Category :
Languages : en
Pages : 254

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Book Description
Since its first experimental demonstration in 1999, Coherent X-Ray Diffractive Imaging has become one of the most promising high resolution X-Ray imaging techniques using coherent radiation produced by brilliant synchrotron storage rings. The ability to directly invert diffraction data with the help of advanced algorithms has paved the way for microscopic investigations and wave-field analyses on the spatial scale of nanometres without the need for inefficient imaging lenses. X-Ray phase contrast which is a measure of the electron density is an important contrast mode of soft biological specimens. For the case of many dominant elements of soft biological matter, the electron density can be converted into an effective mass density offering a unique quantitative information channel which may shed light on important questions such as DNA compaction in the bacterial nucleoid through ‚weighing with light‘. In this work X-Ray phase contrast maps have been obtained from different biological samples by exploring different methods. In particular, the techniques Ptychography and Waveguide-Holographic-Imaging have been used to obtain twodimensional and three-dimensional mass density maps on the single-cell-level of freeze-dried cells of the bacteria Deinococcus radiodurans, Bacillus subtilis and Bacillus thuringiensis allowing, for instance, to estimate the dry weight of the bacterial genome in a near native state. On top of this, reciprocal space information from coherent small angle X-Ray scattering (cellular Nano-Diffraction) of the fine structure of the bacterial cells has been recorded in a synergistic manner and has been analysed down to a resolution of about 2.3/nm exceeding current limits of direct imaging approaches. Furthermore, the dynamic range of present detector technology being one of the major limiting factors of ptychographic phasing of farfield diffraction data has been significantly increased. Overcoming this problem for the case of the very intense X-Ray beam produced by Kirkpatrick-Baez mirrors has been explored by using semi-transparent central stops.

Coherent X-ray diffractive imaging on the single-cell-level of microbial samples

Coherent X-ray diffractive imaging on the single-cell-level of microbial samples PDF Author: Robin Niklas Wilke
Publisher: Göttingen University Press
ISBN: 3863951905
Category :
Languages : en
Pages : 254

Get Book Here

Book Description
Since its first experimental demonstration in 1999, Coherent X-Ray Diffractive Imaging has become one of the most promising high resolution X-Ray imaging techniques using coherent radiation produced by brilliant synchrotron storage rings. The ability to directly invert diffraction data with the help of advanced algorithms has paved the way for microscopic investigations and wave-field analyses on the spatial scale of nanometres without the need for inefficient imaging lenses. X-Ray phase contrast which is a measure of the electron density is an important contrast mode of soft biological specimens. For the case of many dominant elements of soft biological matter, the electron density can be converted into an effective mass density offering a unique quantitative information channel which may shed light on important questions such as DNA compaction in the bacterial nucleoid through ‚weighing with light‘. In this work X-Ray phase contrast maps have been obtained from different biological samples by exploring different methods. In particular, the techniques Ptychography and Waveguide-Holographic-Imaging have been used to obtain twodimensional and three-dimensional mass density maps on the single-cell-level of freeze-dried cells of the bacteria Deinococcus radiodurans, Bacillus subtilis and Bacillus thuringiensis allowing, for instance, to estimate the dry weight of the bacterial genome in a near native state. On top of this, reciprocal space information from coherent small angle X-Ray scattering (cellular Nano-Diffraction) of the fine structure of the bacterial cells has been recorded in a synergistic manner and has been analysed down to a resolution of about 2.3/nm exceeding current limits of direct imaging approaches. Furthermore, the dynamic range of present detector technology being one of the major limiting factors of ptychographic phasing of farfield diffraction data has been significantly increased. Overcoming this problem for the case of the very intense X-Ray beam produced by Kirkpatrick-Baez mirrors has been explored by using semi-transparent central stops.

Coherent X-Ray Diffractive Imaging on the Single-Cell-Level of Microbial Samples

Coherent X-Ray Diffractive Imaging on the Single-Cell-Level of Microbial Samples PDF Author: Robin Niklas Wilke
Publisher:
ISBN:
Category :
Languages : en
Pages :

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Coherent X-Ray Diffractive Imaging on the Single-celll-Level of Microbial Samples

Coherent X-Ray Diffractive Imaging on the Single-celll-Level of Microbial Samples PDF Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 236

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Book Description
Since its first experimental demonstration in 1999, Coherent X-Ray Diffractive Imaging has become one of the most promising high resolution X-Ray imaging techniques using coherent radiation produced by brilliant synchrotron storage rings. The ability to directly invert diffraction data with the help of advanced algorithms has paved the way for microscopic investigations and wave-field analyses on the spatial scale of nanometres without the need for inefficient imaging lenses. X-Ray phase contrast which is a measure of the electron density is an important contrast mode of soft biological spe...

Nanoscale Photonic Imaging

Nanoscale Photonic Imaging PDF Author: Tim Salditt
Publisher: Springer Nature
ISBN: 3030344134
Category : Science
Languages : en
Pages : 634

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Book Description
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.

A Study on New Approaches in Coherent X-ray Microscopy of Biological Specimens

A Study on New Approaches in Coherent X-ray Microscopy of Biological Specimens PDF Author: Klaus Giewekemeyer
Publisher: Universitätsverlag Göttingen
ISBN: 3863950232
Category :
Languages : en
Pages : 226

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Book Description
The use of coherent x rays for microscopic imaging has seen a rapid and ongoing development within the past decade, driven by an increasing availability of highly brilliant and coherent sources worldwide. Accordingly, novel methods have been developed, which replace the microscope‘s objective lens by a numerical reconstruction scheme. The aim of the present work is to study how very recent experimental and algorithmic developments in the field can be implemented towards a highly sensitive and fully quantitative microscopy method for imaging of biological cells. To this end, different experimental approaches are studied, based on coherent far-field as well as near-field diffraction. At first, an application of the novel ptychographic imaging method to single biological cells is presented. In particular, it is demonstrated how weakly scattering biological specimens can be imaged with fully quantitative density contrast. Alongside, a sueccessful extension of the method towards soft x-ray energies is described.In the second part of the work it is shown how x-ray waveguides can be used as a point source for propagation-based microscopy of single cells in the hard x-ray regime. The specifically devised iterative reconstruction scheme allows for full quantitativity and high sensitivity and thus enables an application to single biological cells. The work contains a thorough introduction into the x-ray optical methods applied and aims at a useful and self-contained overview on aspects of signal and Fourier theory relevant for the used numerical propagation schemes.

Coherent X-ray Diffractive Imaging of Biological Samples in 2D and 3D with Synchrotron and XFEL Radiation

Coherent X-ray Diffractive Imaging of Biological Samples in 2D and 3D with Synchrotron and XFEL Radiation PDF Author: Max Rose
Publisher:
ISBN:
Category :
Languages : en
Pages :

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Imaging Whole Escherichia Coli Bacteria by Using Single-Particle X-Ray Diffraction

Imaging Whole Escherichia Coli Bacteria by Using Single-Particle X-Ray Diffraction PDF Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 3

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Book Description
We report the first experimental recording, to our knowledge, of the diffraction pattern from intact Escherichia coli bacteria using coherent x-rays with a wavelength of 2 Å. By using the over-sampling phasing method, a real space image at a resolution of 30 nm was directly reconstructed from the diffraction pattern. An R factor used for characterizing the quality of the reconstruction was in the range of 5%, which demonstrated the reliability of the reconstruction process. The distribution of proteins inside the bacteria labeled with manganese oxide has been identified and this distribution confirmed by fluorescence microscopy images. Compared with lens-based microscopy, this diffraction-based imaging approach can examine thicker samples, such as whole cultured cells, in three dimensions with resolution limited only by radiation damage. Looking forward, the successful recording and reconstruction of diffraction patterns from biological samples reported here represent an important step toward the potential of imaging single biomolecules at near-atomic resolution by combining single-particle diffraction with x-ray free electron lasers.

Multiscale X-Ray Analysis of Biological Cells and Tissues by Scanning Diffraction and Coherent Imaging

Multiscale X-Ray Analysis of Biological Cells and Tissues by Scanning Diffraction and Coherent Imaging PDF Author: Jan-David Nicolas
Publisher: Göttingen University Press
ISBN: 3863954203
Category :
Languages : en
Pages : 183

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Book Description
Understanding the intricate details of muscle contraction has a long-standing tradition in biophysical research. X-ray diffraction has been one of the key techniques to resolve the nanometer-sized molecular machinery involved in force generation. Modern, powerful X-ray sources now provide billions of X-ray photons in time intervals as short as microseconds, enabling fast time-resolved experiments that shed further light on the complex relationship between muscle structure and function. Another approach harnesses this power by repeatedly performing such an experiment at different locations in a sample. With millions of repeated exposures in a single experiment, X-ray diffraction can seamlessly be turned into a raster imaging method, neatly combining real- and reciprocal space information. This thesis has focused on the advancement of this scanning scheme and its application to soft biological tissue, in particular muscle tissue. Special emphasis was placed on the extraction of meaningful, quantitative structural parameters such as the interfilament distance of the actomyosin lattice in cardiac muscle. The method was further adapted to image biological samples on a range of scales, from isolated cells to millimeter-sized tissue sections. Due to the ‘photon-hungry’ nature of the technique, its full potential is often exploited in combination with full-field imaging techniques. From the vast set of microscopic tools available, coherent full-field X-ray imaging has proven to be particularly useful. This multimodal approach allows to correlate two- and three-dimensional images of cells and tissue with diffraction maps of structure parameters. With the set of tools developed in this thesis, scanning X-ray diffraction can now be efficiently used for the structural analysis of soft biological tissues with overarching future applications in biophysical and biomedical research.

Three-dimensional Coherent X-ray Diffractive Imaging of Whole Frozen-hydrated Cells

Three-dimensional Coherent X-ray Diffractive Imaging of Whole Frozen-hydrated Cells PDF Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 9

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X-Ray Diffraction Imaging of Biological Cells

X-Ray Diffraction Imaging of Biological Cells PDF Author: Masayoshi Nakasako
Publisher: Springer
ISBN: 443156618X
Category : Science
Languages : en
Pages : 243

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Book Description
In this book, the author describes the development of the experimental diffraction setup and structural analysis of non-crystalline particles from material science and biology. Recent advances in X-ray free electron laser (XFEL)-coherent X-ray diffraction imaging (CXDI) experiments allow for the structural analysis of non-crystalline particles to a resolution of 7 nm, and to a resolution of 20 nm for biological materials. Now XFEL-CXDI marks the dawn of a new era in structural analys of non-crystalline particles with dimensions larger than 100 nm, which was quite impossible in the 20th century. To conduct CXDI experiments in both synchrotron and XFEL facilities, the author has developed apparatuses, named KOTOBUKI-1 and TAKASAGO-6 for cryogenic diffraction experiments on frozen-hydrated non-crystalline particles at around 66 K. At the synchrotron facility, cryogenic diffraction experiments dramatically reduce radiation damage of specimen particles and allow tomography CXDI experiments. In addition, in XFEL experiments, non-crystalline particles scattered on thin support membranes and flash-cooled can be used to efficiently increase the rate of XFEL pulses. The rate, which depends on the number density of scattered particles and the size of X-ray beams, is currently 20-90%, probably the world record in XFEL-CXDI experiments. The experiment setups and results are introduced in this book. The author has also developed software suitable for efficiently processing of diffraction patterns and retrieving electron density maps of specimen particles based on the diffraction theory used in CXDI.