TY  - JOUR
AU  - Kislinger, Georg
AU  - Fabig, Gunar
AU  - Wehn, Antonia
AU  - Rodriguez, Lucia
AU  - Jiang, Hanyi
AU  - Niemann, Cornelia
AU  - Klymchenko, Andrey S
AU  - Plesnila, Nikolaus
AU  - Misgeld, Thomas
AU  - Müller-Reichert, Thomas
AU  - Khalin, Igor
AU  - Schifferer, Martina
TI  - Combining array tomography with electron tomography provides insights into leakiness of the blood-brain barrier in mouse cortex.
JO  - eLife
VL  - 12
SN  - 2050-084X
CY  - Cambridge
PB  - eLife Sciences Publications
M1  - DZNE-2024-00953
SP  - RP90565
PY  - 2024
AB  - 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.
KW  - Animals
KW  - Mice
KW  - Electron Microscope Tomography: methods
KW  - Blood-Brain Barrier: ultrastructure
KW  - Cerebral Cortex: diagnostic imaging
KW  - Cerebral Cortex: ultrastructure
KW  - Mice, Inbred C57BL
KW  - Male
KW  - Microscopy, Electron, Scanning: methods
KW  - Microtomy
KW  - CLEM (Other)
KW  - array tomography (Other)
KW  - blood brain barrier (Other)
KW  - cell biology (Other)
KW  - correlation (Other)
KW  - imaging (Other)
KW  - mouse (Other)
KW  - neuroscience (Other)
KW  - volume electron microscopy (Other)
LB  - PUB:(DE-HGF)16
C6  - pmid:39102289
C2  - pmc:PMC11299977
DO  - DOI:10.7554/eLife.90565
UR  - https://pub.dzne.de/record/271081
ER  -