title: The CryoCapsule: simplifying correlative light to electron microscopy.
abstract: Correlating complementary multiple scale images of the same object is a straightforward means to decipher biological processes. Light microscopy and electron microscopy are the most commonly used imaging techniques, yet despite their complementarity, the experimental procedures available to correlate them are technically complex. We designed and manufactured a new device adapted to many biological specimens, the CryoCapsule, that simplifies the multiple sample preparation steps, which at present separate live cell fluorescence imaging from contextual high-resolution electron microscopy, thus opening new strategies for full correlative light to electron microscopy. We tested the biological application of this highly optimized tool on three different specimens: the in vitro Xenopus laevis mitotic spindle, melanoma cells over-expressing YFP-langerin sequestered in organized membranous subcellular organelles and a pigmented melanocytic cell in which the endosomal system was labeled with internalized fluorescent transferrin.
authors.affiliation: Institut Curie, Centre de Recherche, Paris, 75248, France; Structure and Membrane Compartments, CNRS UMR144, Paris, 75248, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: Heiligenstein
authors.forename: Jérôme
authors.initials: J
authors.lastname: Heiligenstein
authors.forename: Cédric
authors.initials: C
authors.lastname: Delevoye
authors.forename: Ilse
authors.initials: I
authors.lastname: Hurbain
authors.forename: Sabine
authors.initials: S
authors.lastname: Bardin
authors.forename: Perrine
authors.initials: P
authors.lastname: Paul-Gilloteaux
authors.forename: Lucie
authors.initials: L
authors.lastname: Sengmanivong
authors.forename: Gilles
authors.initials: G
authors.lastname: Régnier
authors.forename: Jean
authors.initials: J
authors.lastname: Salamero
authors.forename: Claude
authors.initials: C
authors.lastname: Antony
authors.forename: Graca
authors.initials: G
authors.lastname: Raposo
mesh_terms: D000818:Animals; D002460:Cell Line; D020285:Cryoelectron Microscopy / Q000295:instrumentation / Q000379:methods*; D016616:Cryoultramicrotomy / Q000295:instrumentation* / Q000379:methods; D004285:Dogs; D011992:Endosomes / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism / Q000648:ultrastructure; D061848:Optical Imaging / Q000295:instrumentation / Q000379:methods; D008941:Spindle Apparatus / Q000378:metabolism / Q000648:ultrastructure; D014981:Xenopus
keywords: CryoCapsule; Xenopus laevis mitotic spindle; correlative light to electron microscopy; endosomal network; freeze substitution; high-pressure freezing; image registration; langerin; melanosomes; spatial resolution; temporal resolution