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
Title Coherent X-ray diffractive imaging on the single-cell-level of microbial samples PDF eBook
Author Robin Niklas Wilke
Publisher Göttingen University Press
Pages 254
Release 2015
Genre
ISBN 3863951905

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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
Title Coherent X-Ray Diffractive Imaging on the Single-Cell-Level of Microbial Samples PDF eBook
Author Robin Niklas Wilke
Publisher
Pages
Release 2014
Genre
ISBN

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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
Title Coherent X-Ray Diffractive Imaging on the Single-celll-Level of Microbial Samples PDF eBook
Author
Publisher
Pages 236
Release 2014
Genre
ISBN

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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
Title Nanoscale Photonic Imaging PDF eBook
Author Tim Salditt
Publisher Springer Nature
Pages 634
Release 2020-06-09
Genre Science
ISBN 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.

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
Title A Study on New Approaches in Coherent X-ray Microscopy of Biological Specimens PDF eBook
Author Klaus Giewekemeyer
Publisher Universitätsverlag Göttingen
Pages 226
Release 2011
Genre
ISBN 3863950232

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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
Title Coherent X-ray Diffractive Imaging of Biological Samples in 2D and 3D with Synchrotron and XFEL Radiation PDF eBook
Author Max Rose
Publisher
Pages
Release 2018
Genre
ISBN

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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
Title Imaging Whole Escherichia Coli Bacteria by Using Single-Particle X-Ray Diffraction PDF eBook
Author
Publisher
Pages 3
Release 2004
Genre
ISBN

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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.