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@ARTICLE{Kislinger:271081,
author = {Kislinger, Georg and Fabig, Gunar and Wehn, Antonia and
Rodriguez, Lucia and Jiang, Hanyi and Niemann, Cornelia and
Klymchenko, Andrey S and Plesnila, Nikolaus and Misgeld,
Thomas and Müller-Reichert, Thomas and Khalin, Igor and
Schifferer, Martina},
title = {{C}ombining array tomography with electron tomography
provides insights into leakiness of the blood-brain barrier
in mouse cortex.},
journal = {eLife},
volume = {12},
issn = {2050-084X},
address = {Cambridge},
publisher = {eLife Sciences Publications},
reportid = {DZNE-2024-00953},
pages = {RP90565},
year = {2024},
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. 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. 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.},
keywords = {Animals / Mice / Electron Microscope Tomography: methods /
Blood-Brain Barrier: ultrastructure / Cerebral Cortex:
diagnostic imaging / Cerebral Cortex: ultrastructure / Mice,
Inbred C57BL / Male / Microscopy, Electron, Scanning:
methods / Microtomy / CLEM (Other) / array tomography
(Other) / blood brain barrier (Other) / cell biology (Other)
/ correlation (Other) / imaging (Other) / mouse (Other) /
neuroscience (Other) / volume electron microscopy (Other)},
cin = {AG Misgeld / AG Simons},
ddc = {600},
cid = {I:(DE-2719)1110000-4 / I:(DE-2719)1110008},
pnm = {351 - Brain Function (POF4-351)},
pid = {G:(DE-HGF)POF4-351},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:39102289},
pmc = {pmc:PMC11299977},
doi = {10.7554/eLife.90565},
url = {https://pub.dzne.de/record/271081},
}