KIT | KIT-Bibliothek | Impressum | Datenschutz

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

Lemercier, Nicolas; Middel, Volker 1; Hentsch, Didier; Taubert, Serge; Takamiya, Masanari 1; Beil, Tanja 1; Vonesch, Jean-Luc; Baumbach, Tilo 2; Schultz, Patrick; Antony, Claude 1; Strähle, Uwe 1
1 Institut für Toxikologie und Genetik (ITG), Karlsruher Institut für Technologie (KIT)
2 Laboratorium für Applikationen der Synchrotronstrahlung (LAS), Karlsruher Institut für Technologie (KIT)


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.

Volltext §
DOI: 10.5445/IR/1000075743
DOI: 10.1038/s41598-017-13348-6
Zitationen: 3
Web of Science
Zitationen: 3
Zitationen: 5
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Toxikologie und Genetik (ITG)
Laboratorium für Applikationen der Synchrotronstrahlung (LAS)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2017
Sprache Englisch
Identifikator ISSN: 2045-2322
KITopen-ID: 1000075743
HGF-Programm 47.01.01 (POF III, LK 01) Biol.Netzwerke u.Synth.Regulat. ITG+ITC
Erschienen in Scientific reports
Verlag Nature Research
Band 7
Seiten Art.Nr.: 13583
Bemerkung zur Veröffentlichung Gefördert durch den KIT-Publikationsfonds
Vorab online veröffentlicht am 19.10.2017
Schlagwörter Cryoelectron tomography; Mechanical engineering; Software; Wide-field fluorescence microscopy
Nachgewiesen in Web of Science
KIT – Die Forschungsuniversität in der Helmholtz-Gemeinschaft
KITopen Landing Page