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A protocol for isolating Xenopus oocyte nuclear envelope for visualization and characterization by scanning electron microscopy (SEM) or transmission electron microscopy (TEM)

Allen, T. D.; Rutherford, S. A.; Murray, S.; Sanderson, Helen; Gardiner, F.; Kiseleva, E.; Goldberg, M. W.; Drummond, S. P.

Authors

T. D. Allen

S. A. Rutherford

S. Murray

F. Gardiner

E. Kiseleva

M. W. Goldberg

S. P. Drummond



Abstract

This protocol details methods for the isolation of oocyte nuclear envelopes (NEs) from the African clawed toad Xenopus laevis, immunogold labeling of component proteins and subsequent visualization by field-emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM). This procedure involves the initial removal of the ovaries from mature female X. laevis, the dissection of individual oocytes, then the manual isolation of the giant nucleus and subsequent preparation for high-resolution visualization. Unlike light microscopy, and its derivative technologies, electron microscopy enables 3-5 nm resolution of nuclear structures, thereby giving unrivalled opportunities for investigation and immunological characterization in situ of nuclear structures and their structural associations. There are a number of stages where samples can be stored, although we recommend that this protocol take no longer than 2 d. Samples processed for FESEM can be stored for weeks under vacuum, allowing considerable time for image acquisition.

Citation

Allen, T. D., Rutherford, S. A., Murray, S., Sanderson, H., Gardiner, F., Kiseleva, E., …Drummond, S. P. (2007). A protocol for isolating Xenopus oocyte nuclear envelope for visualization and characterization by scanning electron microscopy (SEM) or transmission electron microscopy (TEM). Nature Protocols, 2(5), 1166-1172. https://doi.org/10.1038/nprot.2007.137

Journal Article Type Review
Acceptance Date Dec 21, 2006
Online Publication Date May 10, 2007
Publication Date May 1, 2007
Deposit Date Jun 22, 2021
Journal Nature Protocols
Print ISSN 1754-2189
Electronic ISSN 1750-2799
Publisher Nature Research
Peer Reviewed Peer Reviewed
Volume 2
Issue 5
Pages 1166-1172
DOI https://doi.org/10.1038/nprot.2007.137
Public URL https://uwe-repository.worktribe.com/output/7483639