Cryogenic coherent X-ray diffraction imaging of biological samples at SACLA: A correlative approach with cryo-electron and light microscopy

Yuki Takayama, Koji Yonekura

Research output: Contribution to journalReview articlepeer-review

12 Citations (Scopus)

Abstract

Coherent X-ray diffraction imaging at cryogenic temperature (cryo-CXDI) allows the analysis of internal structures of unstained, non-crystalline, whole biological samples in micrometre to sub-micrometre dimensions. Targets include cells and cell organelles. This approach involves preparing frozen-hydrated samples under controlled humidity, transferring the samples to a cryo-stage inside a vacuum chamber of a diffractometer, and then exposing the samples to coherent X-rays. Since 2012, cryo-coherent diffraction imaging (CDI) experiments have been carried out with the X-ray free-electron laser (XFEL) at the SPring-8 Ångstrom Compact free-electron LAser (SACLA) facility in Japan. Complementary use of cryo-electron microscopy and/or light microscopy is highly beneficial for both pre-checking samples and studying the integrity or nature of the sample. This article reports the authors' experience in cryo-XFEL-CDI of biological cells and organelles at SACLA, and describes an attempt towards reliable and higher-resolution reconstructions, including signal enhancement with strong scatterers and Patterson-search phasing.

Original languageEnglish
Pages (from-to)179-189
Number of pages11
JournalActa Crystallographica Section A: Foundations and Advances
Volume72
DOIs
Publication statusPublished - 2016
Externally publishedYes

Keywords

  • coherent X-ray diffraction imaging
  • correlative microscopy
  • frozen-hydrated non-crystalline samples
  • structural analysis
  • X-ray free-electron lasers

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Materials Science(all)
  • Condensed Matter Physics
  • Physical and Theoretical Chemistry
  • Inorganic Chemistry

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