% IMPORTANT: The following is UTF-8 encoded. This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.
@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},
}