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Status: Bibliographieeintrag

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Verfasst von:Wacker, Irene [VerfasserIn]   i
 Veith, Lisa [VerfasserIn]   i
 Spomer, Waldemar [VerfasserIn]   i
 Hofmann, Andreas [VerfasserIn]   i
 Hillmer, Stefan [VerfasserIn]   i
 Gengenbach, Ulrich [VerfasserIn]   i
 Schröder, Rasmus R. [VerfasserIn]   i
Titel:Multimodal hierarchical imaging of serial sections for finding specific cellular targets within large volumes
Verf.angabe:Irene U. Wacker, Lisa Veith, Waldemar Spomer, Andreas Hofmann, Marlene Thaler, Stefan Hillmer, Ulrich Gengenbach, Rasmus R. Schröder
E-Jahr:2018
Jahr:20 March 2018
Umfang:1 Online-Ressource (1 Videodatei, 11:19 min) S.
 9 S.
Illustrationen:farbig
Fussnoten:Enthält auch Textversion ; Gesehen am 11.04.2019 ; Wissenschaftlicher Film. Deutschland. 2018
Titel Quelle:Enthalten in: JoVE. Video journal
Ort Quelle:[S.l.], 2006
Jahr Quelle:2018
Band/Heft Quelle:(2018,133) Artikel-Nummer e57059, 9 Seiten
ISSN Quelle:1940-087X
Abstract:Targeting specific cells at ultrastructural resolution within a mixed cell population or a tissue can be achieved by hierarchical imaging using a combination of light and electron microscopy. Samples embedded in resin are sectioned into arrays consisting of ribbons of hundreds of ultrathin sections and deposited on pieces of silicon wafer or conductively coated coverslips. Arrays are imaged at low resolution using a digital consumer like smartphone camera or light microscope (LM) for a rapid large area overview, or a wide field fluorescence microscope (fluorescence light microscopy (FLM)) after labeling with fluorophores. After post-staining with heavy metals, arrays are imaged in a scanning electron microscope (SEM). Selection of targets is possible from 3D reconstructions generated by FLM or from 3D reconstructions made from the SEM image stacks at intermediate resolution if no fluorescent markers are available. For ultrastructural analysis, selected targets are finally recorded in the SEM at high-resolution (a few nanometer image pixels). A ribbon-handling tool that can be retrofitted to any ultramicrotome is demonstrated. It helps with array production and substrate removal from the sectioning knife boat. A software platform that allows automated imaging of arrays in the SEM is discussed. Compared to other methods generating large volume EM data, such as serial block-face SEM (SBF-SEM) or focused ion beam SEM (FIB-SEM), this approach has two major advantages: (1) The resin-embedded sample is conserved, albeit in a sliced-up version. It can be stained in different ways and imaged with different resolutions. (2) As the sections can be post-stained, it is not necessary to use samples strongly block-stained with heavy metals to introduce contrast for SEM imaging or render the tissue blocks conductive. This makes the method applicable to a wide variety of materials and biological questions. Particularly prefixed materials e.g., from biopsy banks and pathology labs, can directly be embedded and reconstructed in 3D.
DOI:doi:10.3791/57059
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

Volltext: http://dx.doi.org/10.3791/57059
 DOI: https://doi.org/10.3791/57059
Datenträger:Online-Ressource
Dokumenttyp:Film
Sprache:eng
Sach-SW:Humans
 Microscopy, Electron, Scanning
 Microscopy, Fluorescence
 Multimodal Imaging
K10plus-PPN:1663121702
Verknüpfungen:→ Zeitschrift

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