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DOI: 10.5445/IR/1000075743

Microtome-integrated microscope system for high sensitivity tracking of in-resin fluorescence in blocks and ultrathin sections for correlative microscopy

Lemercier, Nicolas; Middel, Volker; Hentsch, Didier; Taubert, Serge; Takamiya, Masanari; Beil, Tanja; Vonesch, Jean-Luc; Baumbach, Tilo; Schultz, Patrick; Antony, Claude; Strähle, Uwe

Abstract:
Many areas of biological research demand the combined use of different imaging modalities to cover a wide range of magnifications and measurements or to place fluorescent patterns into an ultrastructural context. A technically difficult problem is the efficient specimen transfer between different imaging modalities without losing the coordinates of the regions-of-interest (ROI). Here, we report a new and highly sensitive integrated system that combines a custom designed microscope with an ultramicrotome for in-resin-fluorescence detection in blocks, ribbons and sections on EM-grids. Although operating with long-distance lenses, this system achieves a very high light sensitivity. Our instrumental set-up and operating workflow are designed to investigate rare events in large tissue volumes. Applications range from studies of individual immune, stem and cancer cells to the investigation of non-uniform subcellular processes. As a use case, we present the ultrastructure of a single membrane repair patch on a muscle fiber in intact muscle in a whole animal context.


Zugehörige Institution(en) am KIT Institut für Toxikologie und Genetik (ITG)
Laboratorium für Applikationen der Synchrotronstrahlung (LAS)
Publikationstyp Zeitschriftenaufsatz
Jahr 2017
Sprache Englisch
Identifikator ISSN: 2045-2322
URN: urn:nbn:de:swb:90-757439
KITopen ID: 1000075743
HGF-Programm 47.01.01; LK 01
Erschienen in Scientific reports
Band 7
Heft 1
Seiten Art.Nr.: 13583
Bemerkung zur Veröffentlichung Gefördert durch den KIT-Publikationsfonds
Schlagworte Cryoelectron tomography; Mechanical engineering; Software; Wide-field fluorescence microscopy
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