Multiscale ATUM-FIB Microscopy Enables Targeted Ultrastructural Analysis at Isotropic Resolution
2020-07-24 | journal article; research paper
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- Authors
- Kislinger, Georg; Gnägi, Helmut; Kerschensteiner, Martin ; Simons, Mikael ; Misgeld, Thomas ; Schifferer, Martina
- Abstract
- Volume electron microscopy enables the ultrastructural analysis of biological tissue. Currently, the techniques involving ultramicrotomy (ATUM, ssTEM) allow large fields of view but afford only limited z-resolution, whereas ion beam-milling approaches (FIB-SEM) yield isotropic voxels but are restricted in volume size. Now we present a hybrid method, named ATUM-FIB, which combines the advantages of both approaches. ATUM-FIB is based on serial sectioning of tissue into "semithick" (2-10 μm) sections collected onto tape. Serial light and electron microscopy allows the identification of regions of interest that are then directly accessible for targeted FIB-SEM. The set of semithick sections thus represents a tissue "library" which provides three-dimensional context information that can be probed "on demand" by local high-resolution analysis. We demonstrate the potential of this technique to reveal the ultrastructure of rare but pathologically important events by identifying microglia contact sites with amyloid plaques in a mouse model of familial Alzheimer's disease.
- Issue Date
- 24-July-2020
- Journal
- iScience
- Project
- TRR 274: Checkpoints of Central Nervous System Recovery
TRR 274 | B03: Checkpoints of recovery after primary astrocytic lesions in neuromyelitis optica and related animal models
TRR 274 | C02: In vivo detection and targeting of calcium clearance and axonal membrane repair after acute CNS insults
TRR 274 | Z01: Bioimaging Platform - Working Group
- RG Kerschensteiner (Neuroimmune Interactions)
RG Misgeld
RG Schifferer
RG Simons (The Biology of Glia in Development and Disease) - eISSN
- 2589-0042
- Language
- English