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CorRelator: Interactive software for real-time high precision cryo-correlative light and electron microscopy.

Jie E Yang1, Matthew R Larson1, Bryan S Sibert1

  • 1Department of Biochemistry, University of Wisconsin, Madison, WI 53706, United States; Cryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI 53706, United States; Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, WI 53706, United States.

Journal of Structural Biology
|February 21, 2021
PubMed
Summary

CorRelator software enables automated cryo-electron microscopy (cryo-EM) data collection by bridging cryo-fluorescence light microscopy (cryo-FLM) and cryo-electron tomography (cryo-ET). This open-source tool enhances cryo-correlative light and electron microscopy (cryo-CLEM) workflows for high-resolution imaging.

Keywords:
AlgorithmAlignmentCorrelative light and electron microscopyCryo-electron microscopyCryo-electron tomographyCryo-fluorescence microscopy

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biophysics

Background:

  • Cryo-correlative light and electron microscopy (CLEM) combines fluorescence light microscopy (FLM) with cryo-electron microscopy (cryo-EM) for high-resolution imaging.
  • Accurate correlation between FLM and cryo-EM is crucial for identifying cellular structures.
  • Automating cryo-EM data acquisition in cryo-CLEM workflows remains a challenge.

Purpose of the Study:

  • To introduce CorRelator, an open-source software application designed to automate data acquisition in cryo-CLEM.
  • To provide a flexible and accurate method for correlating cryo-FLM and cryo-EM/ET data.
  • To facilitate integration into existing cryo-CLEM workflows across different microscope platforms.

Main Methods:

  • Development of CorRelator, an open-source desktop application with a Graphical User Interface (GUI).
  • Implementation of a pixel-coordinate-to-stage-position transformation for precise correlation.
  • Benchmarking of CorRelator under live-cell and cryogenic conditions using various FLM and TEM instruments.

Main Results:

  • CorRelator successfully bridges cryo-FLM and real-time cryo-EM/ET automated data collection.
  • The software enables flexible, high-accuracy correlation, both on-the-fly and post-acquisition.
  • Demonstrated reliable support for automated correlative cryo-EM/ET acquisition through software-aided and interactive alignment.

Conclusions:

  • CorRelator is a versatile, cross-platform tool that simplifies and automates cryo-CLEM data acquisition.
  • The application enhances the ability to investigate high-resolution ultrastructure by accurately correlating light and electron microscopy data.
  • CorRelator's open-source nature promotes its adoption and integration into diverse cryo-CLEM research.