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| Home > Publications Database > Combining array tomography with electron tomography provides insights into leakiness of the blood-brain barrier in mouse cortex. |
| Home > Publications Database > Combining array tomography with electron tomography provides insights into leakiness of the blood-brain barrier in mouse cortex. |
| Journal Article | DZNE-2024-00953 |
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2024
eLife Sciences Publications
Cambridge
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Please use a persistent id in citations: doi:10.7554/eLife.90565
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.
Keyword(s): Animals (MeSH) ; Mice (MeSH) ; Electron Microscope Tomography: methods (MeSH) ; Blood-Brain Barrier: ultrastructure (MeSH) ; Cerebral Cortex: diagnostic imaging (MeSH) ; Cerebral Cortex: ultrastructure (MeSH) ; Mice, Inbred C57BL (MeSH) ; Male (MeSH) ; Microscopy, Electron, Scanning: methods (MeSH) ; Microtomy (MeSH) ; CLEM ; array tomography ; blood brain barrier ; cell biology ; correlation ; imaging ; mouse ; neuroscience ; volume electron microscopy
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Dataset
Dataset: ATUM-Tomo: A multi-scale approach to cellular ultrastructure by combined volume scanning electron microscopy and electron tomography
BioImage Archive (2024) [10.6019/s-biad1274]
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