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    Visualization of the nucleus and nuclear envelope in situ by SEM in tissue culture cells.

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    Authors
    Allen, Terence D
    Rutherford, Sandra A
    Murray, Stephen M
    Gardiner, Fiona
    Kiseleva, Elena
    Goldberg, Martin W
    Drummond, Sheona P
    Affiliation
    Paterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Withington, Manchester M20 4BX, UK. tallen@picr.man.ac.uk
    Issue Date
    2007
    
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    Abstract
    Our previous work characterizing the biogenesis and structural integrity of the nuclear envelope and nuclear pore complexes (NPCs) has been based on amphibian material but has recently progressed into the analysis of tissue-culture cells. This protocol describes methods for the high resolution visualization, by field-emission scanning electron microscopy (FESEM), of the nucleus and associated structures in tissue culture cells. Imaging by fluorescence light microscopy shows general nuclear and NPC information at a resolution of approximately 200 nm, in contrast to the 3-5 nm resolution provided by FESEM or transmission electron microscopy (TEM), which generates detail at the macromolecular level. The protocols described here are applicable to all tissue culture cell lines tested to date (HeLa, A6, DLD, XTC and NIH 3T3). The processed cells can be stored long term under vacuum. The protocol can be completed in 5 d, including 3 d for cell growth, 1 d for processing and 1 d for imaging.
    Citation
    Visualization of the nucleus and nuclear envelope in situ by SEM in tissue culture cells. 2007, 2 (5):1180-4 Nat Protoc
    Journal
    Nature Protocols
    URI
    http://hdl.handle.net/10541/71224
    DOI
    10.1038/nprot.2007.139
    PubMed ID
    17546013
    Type
    Article
    Language
    en
    ISSN
    1750-2799
    ae974a485f413a2113503eed53cd6c53
    10.1038/nprot.2007.139
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research

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