Title: A novel flat-embedding method to prepare ultrathin cryosections from cultured cells in their in situ orientation
Authors: Oorschot, Viola ×
de Wit, Heidi
Annaert, Wim
Klumperman, Judith #
Issue Date: Aug-2002
Publisher: Histochemical Society
Series Title: Journal of Histochemistry & Cytochemistry vol:50 issue:8 pages:1067-80
Abstract: Immunogold labeling of ultrathin cryosections provides a sensitive and quantitative method to localize proteins at the ultrastructural level. An obligatory step in the routine preparation of cryosections from cultured cells is the detachment of cells from their substrate and subsequent pelleting. This procedure precludes visualization of cells in their in situ orientation and hampers the study of polarized cells. Here we describe a method to sample cultured cells from a petri dish or coverslip by embedding them in a 12% gelatin slab. Subsequently, sections can be prepared in parallel or perpendicular to the plane of growth. Our method extends the cryosectioning technique to applications in studying polarized cells and correlative light-electron microscopy.
ISSN: 0022-1554
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Molecular Genetics Section (-)
Department of Human Genetics - miscellaneous
× corresponding author
# (joint) last author

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