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Title: Three-dimensional coherent X-ray diffractive imaging of whole frozen-hydrated cells

Here, a structural understanding of whole cells in three dimensions at high spatial resolution remains a significant challenge and, in the case of X-rays, has been limited by radiation damage. By alleviating this limitation, cryogenic coherent diffractive imaging (cryo-CDI) can in principle be used to bridge the important resolution gap between optical and electron microscopy in bio-imaging. Here, the first experimental demonstration of cryo-CDI for quantitative three-dimensional imaging of whole frozen-hydrated cells using 8 Kev X-rays is reported. As a proof of principle, a tilt series of 72 diffraction patterns was collected from a frozen-hydrated Neospora caninum cell and the three-dimensional mass density of the cell was reconstructed and quantified based on its natural contrast. This three-dimensional reconstruction reveals the surface and internal morphology of the cell, including its complex polarized sub-cellular structure. Finally, it is believed that this work represents an experimental milestone towards routine quantitative three-dimensional imaging of whole cells in their natural state with spatial resolutions in the tens of nanometres.
Authors:
 [1] ;  [1] ;  [2] ;  [3] ;  [4] ;  [1] ;  [5] ;  [1] ;  [6] ;  [7] ;  [6] ;  [8] ;  [9] ;  [9] ;  [1] ;  [1]
  1. Univ. of California, Los Angeles, CA (United States)
  2. National Sun Yat-sen Univ., Kaohsiung (Taiwan)
  3. Carl ZEISS X-ray Microscopy, Inc., Pleasanton, CA (United States)
  4. Shandong Univ., Jinan (People's Republic of China)
  5. Stanford Univ., Stanford, CA (United States)
  6. Pohang Univ. of Science and Technology, Pohang (South Korea)
  7. Brookhaven National Lab. (BNL), Upton, NY (United States)
  8. European X-ray Free Electron Laser, Hamburg (Germany)
  9. ESRF, Grenoble (France)
Publication Date:
OSTI Identifier:
1261644
Grant/Contract Number:
DARPA-BAA-12-63; GM081409- 01A1; R01#AI064616; 31430031
Type:
Accepted Manuscript
Journal Name:
IUCrJ
Additional Journal Information:
Journal Volume: 2; Journal Issue: 5; Journal ID: ISSN 2052-2525
Publisher:
International Union of Crystallography
Research Org:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org:
USDOE; DARPA; National Inst. of Health (NIH); National Natural Science Foundation of China
Country of Publication:
United States
Language:
English
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE coherent diffractive imaging; cryo-CDI; three-dimensional imaging; three-dimensional cellular structure; coherent diffraction; X-ray imaging; Neospora caninum