Humidity-controlled preparation of frozen-hydrated biological samples for cryogenic coherent x-ray diffraction microscopy

Yuki Takayama, Masayoshi Nakasako

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Coherent x-ray diffraction microscopy (CXDM) has the potential to visualize the structures of micro- to sub-micrometer-sized biological particles, such as cells and organelles, at high resolution. Toward advancing structural studies on the functional states of such particles, here, we developed a system for the preparation of frozen-hydrated biological samples for cryogenic CXDM experiments. The system, which comprised a moist air generator, microscope, micro-injector mounted on a micromanipulator, custom-made sample preparation chamber, and flash-cooling device, allowed for the manipulation of sample particles in the relative humidity range of 20%-94%rh at 293 K to maintain their hydrated and functional states. Here, we report the details of the system and the operation procedure, including its application to the preparation of a frozen-hydrated chloroplast sample. Sample quality was evaluated through a cryogenic CXDM experiment conducted at BL29XUL of SPring-8. Taking the performance of the system and the quality of the sample, the system was suitable to prepare frozen-hydrated biological samples for cryogenic CXDM experiments.

Original languageEnglish
Article number054301
JournalReview of Scientific Instruments
Volume83
Issue number5
DOIs
Publication statusPublished - 2012 May

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Humidity
Cryogenics
cryogenics
humidity
Microscopy
Atmospheric humidity
Microscopic examination
x ray diffraction
Diffraction
X-Rays
microscopy
X rays
preparation
Micromanipulators
Experiments
Chloroplasts
Organelles
Microscopes
chloroplasts
Air

ASJC Scopus subject areas

  • Instrumentation
  • Medicine(all)

Cite this

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