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@MISC{Schifferer:276116,
      author       = {Schifferer, Martina and Rodriguez Martinez, Lucia and
                      Kislinger, Georg and Fabig, Gunar and Wehn, Antonia and
                      Jiang, Hanyi and Niemann, Cornelia and Klymchenko, Andrey S.
                      and Plesnila, Nikolaus and Misgeld, Thomas and
                      Müller-Reichert, Thomas and Khalin, Igor},
      title        = {{D}ataset: {ATUM}-{T}omo: {A} multi-scale approach to
                      cellular ultrastructure by combined volume scanning electron
                      microscopy and electron tomography},
      publisher    = {BioImage Archive},
      reportid     = {DZNE-2025-00197},
      year         = {2024},
      note         = {This work was supported by the DFG under Germany’s
                      Excellence Strategy within the framework of the Munich
                      Cluster for Systems Neurology (SyNergy; EXC 2145 – ID
                      390857198), TRR 274/1 2020 (projects Z01 and B03 – ID
                      408885537) and FOR Immunostroke (Mi 694/9-1 A03 – ID
                      428663564). Research in the Müller-Reichert lab was funded
                      by the DFG (grant MU 1423/8-2 and 8-3 to TMR). All animal
                      experiments were supported by DFG grant 457586042.},
      abstract     = {Like other volume electron microscopy approaches, Automated
                      Tape Collecting Ultramicrotomy (ATUM) enables imaging of
                      serial sections deposited on thick plastic tapes by scanning
                      electron microscopy (SEM). ATUM is unique in enabling
                      hierarchical imaging and thus efficient screening for target
                      structures, as needed for correlative light and electron
                      microscopy. However, SEM of sections on tape can only access
                      the section surface, thereby limiting the axial resolution
                      to the typical size of cellular vesicles with an order of
                      magnitude lower than the acquired xy resolution. In
                      contrast, serial-section electron tomography (ET), a
                      transmission electron microscopy-based approach, yields
                      isotropic voxels at full EM resolution, but requires
                      deposition of sections on electron-stable thin and fragile
                      films, thus making screening of large section libraries
                      difficult and prone to section loss. To combine the strength
                      of both approaches, we developed ‘ATUM-Tomo’, a hybrid
                      method, where sections are first reversibly attached to
                      plastic tape via a dissolvable coating, and after screening
                      detached and transferred to the ET-compatible thin films.
                      ATUM-SEM of serial semi-thick sections and consecutive ET of
                      selected re-mounted sections combines SEM’s fast target
                      recognition and coarse rendering capability with
                      high-resolution volume visualizations, respectively, thus
                      enabling multi-scale interrogation of cellular
                      ultrastructure. As a proof-of-principle, we applied
                      correlative ATUM-Tomo to study ultrastructural features of
                      blood brain barrier (BBB) leakiness around microthrombi in a
                      mouse model of traumatic brain injury. Microthrombi and
                      associated sites of BBB leakiness were identified by
                      confocal imaging of injected fluorescent and electron-dense
                      nanoparticles, then relocalized by ATUM-SEM, and finally
                      interrogated by correlative ATUM-Tomo. This workflow is a
                      seamless zoom-in on structural BBB pathology from the micro-
                      to the nanometer scale. Overall, our new ATUM-Tomo approach
                      will substantially advance ultrastructural analysis of
                      biological phenomena that require cell- and tissue-level
                      contextualization of the finest subcellular textures.},
      cin          = {AG Misgeld},
      cid          = {I:(DE-2719)1110000-4},
      pnm          = {351 - Brain Function (POF4-351) / DFG project
                      G:(GEPRIS)390857198 - EXC 2145: Munich Cluster for Systems
                      Neurology (SyNergy) (390857198)},
      pid          = {G:(DE-HGF)POF4-351 / G:(GEPRIS)390857198},
      typ          = {PUB:(DE-HGF)32},
      doi          = {10.6019/s-biad1274},
      url          = {https://pub.dzne.de/record/276116},
}