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

Jie E Yang; Matthew R Larson; Bryan S Sibert; Samantha Shrum; Elizabeth R Wright

doi:10.1016/j.jsb.2021.107709

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

J Struct Biol. 2021 Jun;213(2):107709. doi: 10.1016/j.jsb.2021.107709. Epub 2021 Feb 18.

Authors

Jie E Yang¹, Matthew R Larson¹, Bryan S Sibert¹, Samantha Shrum², Elizabeth R Wright³

Affiliations

¹ Department 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.
² Biophysics Graduate Program, University of Wisconsin, Madison, WI 53706, United States.
³ Department 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; Biophysics Graduate Program, University of Wisconsin, Madison, WI 53706, United States; Morgridge Institute for Research, Madison, WI, 53715, United States; Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, WI 53706, United States. Electronic address: erwright2@wisc.edu.

Abstract

Cryo-correlative light and electron microscopy (CLEM) is a technique that uses the spatiotemporal cues from fluorescence light microscopy (FLM) to investigate the high-resolution ultrastructure of biological samples by cryo-electron microscopy (cryo-EM). Cryo-CLEM provides advantages for identifying and distinguishing fluorescently labeled proteins, macromolecular complexes, and organelles from the cellular environment. Challenges remain on how correlation workflows and software tools are implemented on different microscope platforms to support automated cryo-EM data acquisition. Here, we present CorRelator: an open-source desktop application that bridges between cryo-FLM and real-time cryo-EM/ET automated data collection. CorRelator implements a pixel-coordinate-to-stage-position transformation for flexible, high accuracy on-the-fly and post-acquisition correlation. CorRelator can be integrated into cryo-CLEM workflows and easily adapted to standard fluorescence and transmission electron microscope (TEM) system configurations. CorRelator was benchmarked under live-cell and cryogenic conditions using several FLM and TEM instruments, demonstrating that CorRelator reliably supports real-time, automated correlative cryo-EM/ET acquisition, through a combination of software-aided and interactive alignment. CorRelator is a cross-platform software package featuring an intuitive Graphical User Interface (GUI) that guides the user through the correlation process. CorRelator source code is available at: https://github.com/wright-cemrc-projects/corr.

Keywords: Algorithm; Alignment; Correlative light and electron microscopy; Cryo-electron microscopy; Cryo-electron tomography; Cryo-fluorescence microscopy.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cryoelectron Microscopy*
Electron Microscope Tomography
HeLa Cells
Humans
Image Processing, Computer-Assisted*
Microscopy, Fluorescence*
Respiratory Syncytial Virus Infections / pathology
Software*
User-Computer Interface
Workflow

Abstract

Publication types

MeSH terms

Grants and funding