Scanning X-Ray Nanodiffraction on Dictyostelium discoideum

Priebe, Marius; Bernhardt, Marten; Blum, Christoph; Tarantola, Marco; Bodenschatz, Eberhard; Salditt, Tim

doi:10.1016/j.bpj.2014.10.027

Cited by 38 publications

(118 citation statements)

References 36 publications

(51 reference statements)

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“…Independent of any labeling such as for fluorescence-based microscopy, scanning SAXS detects all cellular constituents present in proportion to the respective electron density contrast and distribution. For cellular experiments, structural attributes down to an equivalent structure size of a few nanometers can thus be detected [7,10,11].…”

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confidence: 99%

Anisotropic x-ray scattering and orientation fields in cardiac tissue cells

et al. 2017

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X-ray diffraction from biomolecular assemblies is a powerful technique which can provide structural information about complex architectures such as the locomotor systems underlying muscle contraction. However, in its conventional form, macromolecular diffraction averages over large ensembles. Progress in x-ray optics has now enabled to probe structures on sub-cellular scales, with the beam confined to a distinct organelle. Here, we use scanning small angle x-ray scattering (scanning SAXS) to probe the diffraction from cytoskeleton networks in cardiac tissue cells. In particular, we focus on actin-myosin composites, which we identify as the dominating contribution to the anisotropic diffraction patterns, by correlation with optical fluorescence microscopy. To this end, we use a principal component analysis approach to quantify direction, degree of orientation, nematic order, and the second moment of the scattering distribution in each scan point. We compare the fiber orientation from micrographs of fluorescently labeled actin fibers to the structure orientation of the x-ray dataset and thus correlate signals of two different measurements: the native electron density distribution of the local probing area versus specifically labeled constituents of the sample. Further, we develop a robust and automated fitting approach based on a power law expansion, in order to describe the local structure factor in each scan point over a broad range of the momentum transfer q r . Finally, we demonstrate how the methodology shown for freeze dried cells in the first part of the paper can be translated to alive cell recordings.Recent progress in x-ray optics has now overcome this barrier, enabling hard x-ray spot sizes in the submicron range [24], well suited to record structural data within precise locations of a single cell. X-rays even in the multi-keV regime required for diffraction studies can nowadays be focussed by a variety of optical elements, including diffractive optics such as Fresnel zone plates [25], compound refractive lenses [26-28] and elliptical Kirkpatrick-Baez (KB) mirrors [29][30][31][32]. Similar to earlier scanning diffraction work on biomaterials such as wood and bone [33][34][35][36][37][38], we can hence now combine high resolution in reciprocal space with at least moderate resolution in real space. Scanning small angle x-ray scattering (scanning SAXS) experiments requiring a sample environment for biological cells are typically not compatible with the ultimate small spot sizes of 10 nm and below, as presented in [31,39,40], but values in the range of 80-300 nm are feasible, in particular in terms of the working distance, and readily allow structure factors to be assigned to different cellular compartments. Presently, feasibility of cellular scanning SAXS has been demonstrated for a variety of biological cells, ranging from bacterial cells D. radiodurans [6] to eukaryotes such as the amoeba D. discoideum [10], adenoma cells [7-9, 41], and human mesenchymal stem cells (hMSC) [11].Beyond these previous proof-...

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Anisotropic x-ray scattering and orientation fields in cardiac tissue cells

et al. 2017

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“…Firstly, it 188 Summary, Conclusion & Outlook represents the prototype of a scanning (coherent) SAXS experiment on the level of single cells that was also applied in recent works such as [105,240,323]. Presently, the obtained resolution in the range of about 2 nm −1 in reciprocal space 27 exceeds any achieved real-space resolution on unstained cellular samples.…”

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confidence: 98%

“…[65, 130, 200, 201, 264 266, 313, 314]). Scanning SAXS experiments using these nano-beams are ideally suited to analyse melanosomes and eukaryotic cells, even in a fully hydrated and living state [104,105,239,240,322,323].…”

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

Wilke

2015

Göttingen Series in X-Ray Physics

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“…Also, further analysis routines such as the automized detection of diffracton streaks typical for fibre scattering (streak-finder algorithm, [3]), Fresnel propagation and (holographic) phase retrieval methods, as well as rudimentary tomographic functions (sinogram generation, filtered back projection) will be added as analysis modules.…”

Section: Discussionmentioning

confidence: 99%

dada – a web-based 2D detector analysis tool

Osterhoff

2017

J. Phys.: Conf. Ser.

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Abstract. The data daemon, dada, is a server backend for unified access to 2D pixel detector image data stored with different detectors, file formats and saved with varying naming conventions and folder structures across instruments. Furthermore, dada implements basic preprocessing and analysis routines from pixel binning over azimuthal integration to raster scan processing. Common user interactions with dada are by a web frontend, but all parameters for an analysis are encoded into a Uniform Resource Identifier (URI) which can also be written by hand or scripts for batch processing.

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Scanning X-Ray Nanodiffraction on Dictyostelium discoideum

Cited by 38 publications

References 36 publications

Anisotropic x-ray scattering and orientation fields in cardiac tissue cells

Anisotropic x-ray scattering and orientation fields in cardiac tissue cells

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

dada – a web-based 2D detector analysis tool

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