Microtome-integrated microscope system for high sensitivity tracking of in-resin fluorescence in blocks and ultrathin sections for correlative microscopy.
Fiche publication
Date publication
octobre 2017
Journal
Scientific reports
Auteurs
Membres identifiés du Cancéropôle Est :
Dr SCHULTZ Patrick, Dr VONESCH Jean-Luc, Dr LEMERCIER Nicolas
Tous les auteurs :
Lemercier N, Middel V, Hentsch D, Taubert S, Takamiya M, Beil T, Vonesch JL, Baumbach T, Schultz P, Antony C, Strähle U
Lien Pubmed
Résumé
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.
Référence
Sci Rep. 2017 Oct 19;7(1):13583