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Title: Procedures for cryogenic X-ray ptychographic imaging of biological samples

Abstract

Biological sample-preparation procedures have been developed for imaging human chromosomes under cryogenic conditions. A new experimental setup, developed for imaging frozen samples using beamline I13 at Diamond Light Source, is described. This paper describes the equipment and experimental procedures as well as the authors' first ptychographic reconstructions using X-rays.

Authors:
 [1];  [2];  [3];  [3];  [4];  [5];  [5];  [3];  [6]
  1. Univ. College London, London (United Kingdom). London Centre for Nanotechnology; Research Complex at Harwell, Oxfordshire (England). Rutherford Appleton Lab.; Brunel Univ. London, Uxbridge (England)
  2. Univ. College London, London (United Kingdom). London Centre for Nanotechnology; Research Complex at Harwell, Oxfordshire (England). Rutherford Appleton Lab.; Southern Univ. of Science and Technology, Shenzhen (China)
  3. Univ. College London, London (United Kingdom). London Centre for Nanotechnology; Research Complex at Harwell, Oxfordshire (England). Rutherford Appleton Lab.
  4. Research Complex at Harwell, Oxfordshire (England). Rutherford Appleton Lab.
  5. Diamond Light Source, Didcot, Oxfordshire (England)
  6. Univ. College London, London (United Kingdom). London Centre for Nanotechnology; Research Complex at Harwell, Oxfordshire (England). Rutherford Appleton Lab.; Brookhaven National Lab. (BNL), Upton, NY (United States). Condensed Matter Physics and Materials Science Dept.
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1339020
Alternate Identifier(s):
OSTI ID: 1342640
Report Number(s):
BNL-113441-2017-JA
Journal ID: ISSN 2052-2525; IUCRAJ; R&D Project: PO011; KC0201060; TRN: US1700989
Grant/Contract Number:
SC0012704; BB/ H022597/1; SC00112704
Resource Type:
Journal Article: Published Article
Journal Name:
IUCrJ
Additional Journal Information:
Journal Volume: 4; Journal Issue: 2; Journal ID: ISSN 2052-2525
Publisher:
International Union of Crystallography
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 47 OTHER INSTRUMENTATION; human chromosomes; cell nuclei; chromosome imaging; X-ray ptychography; plunge freezing

Citation Formats

Yusuf, M., Zhang, F., Chen, B., Bhartiya, A., Cunnea, K., Wagner, U., Cacho-Nerin, F., Schwenke, J., and Robinson, I. K. Procedures for cryogenic X-ray ptychographic imaging of biological samples. United States: N. p., 2017. Web. doi:10.1107/S2052252516020029.
Yusuf, M., Zhang, F., Chen, B., Bhartiya, A., Cunnea, K., Wagner, U., Cacho-Nerin, F., Schwenke, J., & Robinson, I. K. Procedures for cryogenic X-ray ptychographic imaging of biological samples. United States. doi:10.1107/S2052252516020029.
Yusuf, M., Zhang, F., Chen, B., Bhartiya, A., Cunnea, K., Wagner, U., Cacho-Nerin, F., Schwenke, J., and Robinson, I. K. Thu . "Procedures for cryogenic X-ray ptychographic imaging of biological samples". United States. doi:10.1107/S2052252516020029.
@article{osti_1339020,
title = {Procedures for cryogenic X-ray ptychographic imaging of biological samples},
author = {Yusuf, M. and Zhang, F. and Chen, B. and Bhartiya, A. and Cunnea, K. and Wagner, U. and Cacho-Nerin, F. and Schwenke, J. and Robinson, I. K.},
abstractNote = {Biological sample-preparation procedures have been developed for imaging human chromosomes under cryogenic conditions. A new experimental setup, developed for imaging frozen samples using beamline I13 at Diamond Light Source, is described. This paper describes the equipment and experimental procedures as well as the authors' first ptychographic reconstructions using X-rays.},
doi = {10.1107/S2052252516020029},
journal = {IUCrJ},
number = 2,
volume = 4,
place = {United States},
year = {Thu Jan 12 00:00:00 EST 2017},
month = {Thu Jan 12 00:00:00 EST 2017}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1107/S2052252516020029

Citation Metrics:
Cited by: 6works
Citation information provided by
Web of Science

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  • Biological sample-preparation procedures have been developed for imaging human chromosomes under cryogenic conditions. A new experimental setup, developed for imaging frozen samples using beamline I13 at Diamond Light Source, is described. This paper describes the equipment and experimental procedures as well as the authors' first ptychographic reconstructions using X-rays.
  • Bionanoprobe (BNP), a hard x-ray fluorescence sample-scanning nanoprobe at the Advanced Photon Source of Argonne National Laboratory, has been used to quantitatively study elemental distributions in biological cells with sub-100 nm spatial resolution and high sensitivity. Cryogenic conditions enable biological samples to be studied in their frozen-hydrated state with both ultrastructure and elemental distributions more faithfully preserved compared to conventional chemical fixation or dehydration methods. Furthermore, radiation damage is reduced in two ways: the diffusion rate of free radicals is decreased at low temperatures; and the sample is embedded in vitrified ice, which reduces mass loss.
  • Coherent X-ray diffraction imaging (CXDI) is a technique for structure analyses of non-crystalline particles with dimensions ranging from micrometer to sub-micrometer. We have developed a diffraction apparatus named TAKASAGO-6 for use in single-shot CXDI experiments of frozen-hydrated non-crystalline biological particles at cryogenic temperature with X-ray free electron laser pulses provided at a repetition rate of 30 Hz from the SPring-8 Angstrom Compact free-electron LAser. Specimen particles are flash-cooled after being dispersed on thin membranes supported by specially designed disks. The apparatus is equipped with a high-speed translation stage with a cryogenic pot for raster-scanning of the disks at a speedmore » higher than 25 μm/33 ms. In addition, we use devices assisting the easy transfer of cooled specimens from liquid-nitrogen storages to the cryogenic pot. In the current experimental procedure, more than 20 000 diffraction patterns can be collected within 1 h. Here we report the key components and performance of the diffraction apparatus. Based on the efficiency of the diffraction data collection and the structure analyses of metal particles, biological cells, and cellular organelles, we discuss the future application of this diffraction apparatus for structure analyses of biological specimens.« less
  • X-ray diffraction microscopy (XDM) is well suited for nondestructive, high-resolution biological imaging, especially for thick samples, with the high penetration power of x rays and without limitations imposed by a lens. We developed nonvacuum, cryogenic (cryo-) XDM with hard x rays at 8 keV and report the first frozen-hydrated imaging by XDM. By preserving samples in amorphous ice, the risk of artifacts associated with dehydration or chemical fixation is avoided, ensuring the imaging condition closest to their natural state. The reconstruction shows internal structures of intact D. radiodurans bacteria in their natural contrast.