Imaging of Biological Materials and Cells by X-ray Scattering and Diffraction

Hémonnot, Clément Y. J.; Köster, Sarah

doi:10.1021/acsnano.7b03447

Cited by 70 publications

(58 citation statements)

References 173 publications

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“…Recently, however, we have increasing knowledge about how to preserve or recover the phase information in X-ray diffraction experiments, which is owed to advanced algorithms and improved computer performance. The techniques to restore the real-space images of the object by using this knowledge are collectively called coherent diffractive imaging (CDI) or lens less imaging [ 56 , 57 , 58 ]. To do this, coherent illumination of objects is required.…”

Section: Future Directionsmentioning

confidence: 99%

Synchrotron Radiation X-ray Diffraction Techniques Applied to Insect Flight Muscle

Iwamoto

2018

IJMS

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X-ray fiber diffraction is a powerful tool used for investigating the molecular structure of muscle and its dynamics during contraction. This technique has been successfully applied not only to skeletal and cardiac muscles of vertebrates but also to insect flight muscle. Generally, insect flight muscle has a highly ordered structure and is often capable of high-frequency oscillations. The X-ray diffraction studies on muscle have been accelerated by the advent of 3rd-generation synchrotron radiation facilities, which can generate brilliant and highly oriented X-ray beams. This review focuses on some of the novel experiments done on insect flight muscle by using synchrotron radiation X-rays. These include diffraction recordings from single myofibrils within a flight muscle fiber by using X-ray microbeams and high-speed diffraction recordings from the flight muscle during the wing-beat of live insects. These experiments have provided information about the molecular structure and dynamic function of flight muscle in unprecedented detail. Future directions of X-ray diffraction studies on muscle are also discussed.

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Section: Future Directionsmentioning

confidence: 99%

Synchrotron Radiation X-ray Diffraction Techniques Applied to Insect Flight Muscle

Iwamoto

2018

IJMS

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“…Thanks to super-resolution techniques (Hell, 2007), fluorescence microscopy is widely used in labeled, intact cells (Ferná ndez-Suá rez & Ting, 2008;Huang et al, 2010) and can resolve details on the order of tens of nanometers. X-ray imaging techniques (Kirz et al, 1995;Hé monnot & Kö ster, 2017) rely on the small wavelength and high penetration depth of X-radiation. In particular, scanning small-angle X-ray scattering (SAXS) (Fratzl et al, 1997) is used on unsliced, unstained samples to obtain both real and reciprocal space information.…”

Section: Introductionmentioning

confidence: 99%

Large field-of-view scanning small-angle X-ray scattering of mammalian cells

Cassini¹,

Wittmeier²,

Brehm³

et al. 2020

J Synchrotron Rad

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X-ray imaging is a complementary method to electron and fluorescence microscopy for studying biological cells. In particular, scanning small-angle X-ray scattering provides overview images of whole cells in real space as well as local, high-resolution reciprocal space information, rendering it suitable to investigate subcellular nanostructures in unsliced cells. One persisting challenge in cell studies is achieving high throughput in reasonable times. To this end, a fast scanning mode is used to image hundreds of cells in a single scan. A way of dealing with the vast amount of data thus collected is suggested, including a segmentation procedure and three complementary kinds of analysis, i.e. characterization of the cell population as a whole, of single cells and of different parts of the same cell. The results show that short exposure times, which enable faster scans and reduce radiation damage, still yield information in agreement with longer exposure times.

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“…This enables recordings of the small-angle x-ray scattering (SAXS) while scanning the sample in real space, as initially introduced for biomaterials (11,12), more recently also for soft matter (13), as well as for 3d vector SAXS (14). Scanning diffraction from biological cells with spot sizes smaller than a single organelle are still more challenging in terms of signal-to-noise (15)(16)(17)(18)(19)(20), as well as radiation damage (21). Recently, we have used this approach to study the cytoskeletal structure of single CMs in order to compare the signal level for different preparation states (22).…”

Section: Introductionmentioning

confidence: 99%

X-ray structural analysis of single adult cardiomyocytes: tomographic imaging and micro-diffraction

Reichardt¹,

Neuhaus²,

Nicolas³

et al. 2020

Preprint

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ABSTRACTWe present a multi-scale imaging approach to characterize the structure of isolated adult murine cardiomyocytes based on a combination of full-field three-dimensional (3d) coherent x-ray imaging and scanning x-ray diffraction. Using these modalities, we probe the structure from the molecular to the cellular scale. Holographic projection images on freeze-dried cells have been recorded using highly coherent and divergent x-ray waveguide radiation. Phase retrieval and tomographic reconstruction then yield the 3d electron density distribution with a voxel size below 50 nm. In the reconstruction volume, myofibrils, sarcomeric organisation and mitochondria can be visualized and quantified within a single cell without sectioning. Next, we use micro-focusing optics by compound refractive lenses to probe the diffraction signal of the acto-myosin lattice. Comparison between recordings of chemically fixed and untreated, living cells indicate that the characteristic lattice distances shrink by approximately 10% upon fixation.SIGNIFICANCEDiffraction with synchrotron radiation has played an important role to decipher the molecular structure underlying force generation in muscle. In this work, the diffraction signal of the actomyosin contractile unit has for the first time been recorded from living cardiomyocytes, bringing muscle diffraction to the scale of single cells. In addition to scanning diffraction, we use coherent optics at the same synchrotron endstation to perform holographic imaging and tomography on a single cardiomyocyte. By this hard x-ray microscopy modality, we extend the length scales covered by scanning diffraction and reconstruct the electron density of an entire freeze-dried cardiomyocyte, visualizing the 3d architecture of myofibrils, sarcomers, and mitochondria with a voxel size below 50 nm.

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Imaging of Biological Materials and Cells by X-ray Scattering and Diffraction

Cited by 70 publications

References 173 publications

Synchrotron Radiation X-ray Diffraction Techniques Applied to Insect Flight Muscle

Synchrotron Radiation X-ray Diffraction Techniques Applied to Insect Flight Muscle

Large field-of-view scanning small-angle X-ray scattering of mammalian cells

X-ray structural analysis of single adult cardiomyocytes: tomographic imaging and micro-diffraction

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