<p>Volume correlative light and electron microscopy (vCLEM) is a powerful imaging technique that enables the visualization of fluorescently labeled proteins within their ultrastructural context. Currently, vCLEM alignment relies on time-consuming and subjective manual methods. This paper presents CLEM-Reg, an algorithm that automates the three-dimensional alignment of vCLEM datasets by leveraging probabilistic point cloud registration techniques. Point clouds are derived from segmentations of common structures in each modality, created by state-of-the-art open-source methods. CLEM-Reg drastically reduces the registration time of vCLEM datasets to a few minutes and achieves correlation of fluorescent signal to submicron target structures in electron microscopy on three newly acquired vCLEM benchmark datasets. CLEM-Reg was then used to automatically obtain vCLEM overlays to unambiguously identify TGN46-positive transport carriers involved in protein trafficking between the trans-Golgi network and plasma membrane. Datasets are available on EMPIAR and BioStudies, and a napari plugin is provided to aid end-user adoption.</p>

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CLEM-Reg: an automated point cloud-based registration algorithm for volume correlative light and electron microscopy

  • Daniel Krentzel,
  • Matouš Elphick,
  • Marie-Charlotte Domart,
  • Christopher J. Peddie,
  • Romain F. Laine,
  • Cameron Shand,
  • Ricardo Henriques,
  • Lucy M. Collinson,
  • Martin L. Jones

摘要

Volume correlative light and electron microscopy (vCLEM) is a powerful imaging technique that enables the visualization of fluorescently labeled proteins within their ultrastructural context. Currently, vCLEM alignment relies on time-consuming and subjective manual methods. This paper presents CLEM-Reg, an algorithm that automates the three-dimensional alignment of vCLEM datasets by leveraging probabilistic point cloud registration techniques. Point clouds are derived from segmentations of common structures in each modality, created by state-of-the-art open-source methods. CLEM-Reg drastically reduces the registration time of vCLEM datasets to a few minutes and achieves correlation of fluorescent signal to submicron target structures in electron microscopy on three newly acquired vCLEM benchmark datasets. CLEM-Reg was then used to automatically obtain vCLEM overlays to unambiguously identify TGN46-positive transport carriers involved in protein trafficking between the trans-Golgi network and plasma membrane. Datasets are available on EMPIAR and BioStudies, and a napari plugin is provided to aid end-user adoption.