Multiscale ATUM-FIB Microscopy Enables Targeted Ultrastructural Analysis at Isotropic Resolution.

Biological Sciences Cellular Neuroscience Experimental Systems for Structural Biology Imaging Anatomy

Journal

iScience
ISSN: 2589-0042
Titre abrégé: iScience
Pays: United States
ID NLM: 101724038

Informations de publication

Date de publication:
24 Jul 2020
Historique:
received: 06 04 2020
revised: 11 05 2020
accepted: 15 06 2020
pubmed: 6 7 2020
medline: 6 7 2020
entrez: 5 7 2020
Statut: ppublish

Résumé

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.

Identifiants

pubmed: 32622266
pii: S2589-0042(20)30477-6
doi: 10.1016/j.isci.2020.101290
pmc: PMC7334410
pii:
doi:

Types de publication

Journal Article

Langues

eng

Pagination

101290

Subventions

Organisme : European Research Council
ID : 616791
Pays : International

Informations de copyright

Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of Interests The authors declare no competing interest.

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Auteurs

Georg Kislinger (G)

German Center for Neurodegenerative Diseases (DZNE), Munich 81377, Germany; Munich Cluster of Systems Neurology (SyNergy), Munich 81377, Germany; Institute of Neuronal Cell Biology, Technical University Munich, Munich 80802, Germany.

Helmut Gnägi (H)

Diatome SA, Helmstrasse 1, 2560 Nidau, Switzerland.

Martin Kerschensteiner (M)

Munich Cluster of Systems Neurology (SyNergy), Munich 81377, Germany; Institute of Clinical Neuroimmunology, University Hospital, Ludwig-Maximilians-University Munich, Planegg-Martinsried 82152, Germany; Biomedical Center (BMC), Faculty of Medicine, Ludwig-Maximilians-University Munich, Planegg-Martinsried 82152, Germany.

Mikael Simons (M)

German Center for Neurodegenerative Diseases (DZNE), Munich 81377, Germany; Munich Cluster of Systems Neurology (SyNergy), Munich 81377, Germany; Institute of Neuronal Cell Biology, Technical University Munich, Munich 80802, Germany.

Thomas Misgeld (T)

German Center for Neurodegenerative Diseases (DZNE), Munich 81377, Germany; Munich Cluster of Systems Neurology (SyNergy), Munich 81377, Germany; Institute of Neuronal Cell Biology, Technical University Munich, Munich 80802, Germany.

Martina Schifferer (M)

German Center for Neurodegenerative Diseases (DZNE), Munich 81377, Germany; Munich Cluster of Systems Neurology (SyNergy), Munich 81377, Germany. Electronic address: martina.schifferer@dzne.de.

Classifications MeSH