Home Cell Biology Fully Autonomous Characterization and Data Collection from Crystals of Biological Macromolecules
Cell Biology JoVE (Open Access) Citable · DOI

Fully Autonomous Characterization and Data Collection from Crystals of Biological Macromolecules

DOI: 10.3791/59032-v
What you'll learn
  • Set up automated diffraction screening and data collection at synchrotron beamlines
  • Create and manage diffraction plans using ISPyB software
  • Retrieve and interpret processed X-ray crystallography datasets without manual intervention
Protocol

Here, we describe how to use the automated screening and data collection options available at some synchrotron beamlines. Scientists send cryocooled samples to the synchrotron, and the diffraction properties are screened, the data sets are collected and processed and, where possible, a structure solution is carried out—all without human intervention.

Difficulty
advanced
Total time
~3–7 days (sample preparation + beamline access + automated processing)

Steps

1
Create diffraction plan in ISPyB

Design and configure automated screening and data collection parameters using the ISPyB software interface. This defines the workflow for unattended sample characterization and dataset acquisition.

▶ 00:45
2
Execute data collection and retrieve results

Initiate autonomous data collection at the synchrotron beamline, monitor data processing in real-time, and download processed diffraction datasets and structure solutions.

▶ 03:55
3
Analyze X-ray diffraction data

Examine processed crystallographic data and interpret structure solutions obtained from automated analysis, exemplified by human mitochondrial GCSH protein characterization.

▶ 05:02
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