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dc.contributor.authorGoldberg, Martin W
dc.contributor.authorBlow, J J
dc.contributor.authorAllen, Terence D
dc.date.accessioned2010-08-03T08:44:37Z
dc.date.available2010-08-03T08:44:37Z
dc.date.issued2010-08-03T08:44:37Z
dc.identifier.citationThe use of field emission in-lens scanning electron microscopy to study the steps of assembly of the nuclear envelope in vitro., 108 (3):257-68 J. Struct. Biol.en
dc.identifier.issn1047-8477
dc.identifier.pmid1476831
dc.identifier.doi10.1016/1047-8477(92)90026-7
dc.identifier.urihttp://hdl.handle.net/10541/108885
dc.description.abstractAt mitosis the nuclear envelope (NE) is disassembled to allow chromosome separation. In telophase it is reassembled as the chromosomes decondense. Cell-free extracts of Xenopus eggs have been used extensively to study assembly of the NE and the nuclear pore complexes (NPCs), providing several models for the steps involved. The NE is a surface structure which in cell-free extracts is easily exposed. It is appropriate, therefore, to use a surface imaging technique to study NE dynamics. Field emission in-lens scanning electron microscopy (FEISEM) provides the opportunity to image surfaces, directly, and to visualise details of structures such as the NPC. Here we show the feasibility and value of FEISEM to study the steps of NE formation. Nuclei have been assembled in vitro and fixed at different time points during assembly, followed by conductive staining, platinum coating, and visualisation by FEISEM. Changes on the nuclear surface with time are shown. Details of the surface of chromatin and the cytoplasmic face of NPC structure are demonstrated without the need to isolate the structures from the nucleus.
dc.language.isoenen
dc.subject.meshAnimals
dc.subject.meshChromatin
dc.subject.meshEvaluation Studies as Topic
dc.subject.meshFemale
dc.subject.meshMale
dc.subject.meshMicroscopy, Electron, Scanning
dc.subject.meshNuclear Envelope
dc.subject.meshOocytes
dc.subject.meshSperm Head
dc.subject.meshXenopus laevis
dc.titleThe use of field emission in-lens scanning electron microscopy to study the steps of assembly of the nuclear envelope in vitro.en
dc.typeArticleen
dc.contributor.departmentCRC Department of Structural Cell Biology, Paterson Institute for Cancer Research, Christie Hospital, Holt Radium Institute, Manchester, United Kingdom.en
dc.identifier.journalJournal of Structural Biologyen
html.description.abstractAt mitosis the nuclear envelope (NE) is disassembled to allow chromosome separation. In telophase it is reassembled as the chromosomes decondense. Cell-free extracts of Xenopus eggs have been used extensively to study assembly of the NE and the nuclear pore complexes (NPCs), providing several models for the steps involved. The NE is a surface structure which in cell-free extracts is easily exposed. It is appropriate, therefore, to use a surface imaging technique to study NE dynamics. Field emission in-lens scanning electron microscopy (FEISEM) provides the opportunity to image surfaces, directly, and to visualise details of structures such as the NPC. Here we show the feasibility and value of FEISEM to study the steps of NE formation. Nuclei have been assembled in vitro and fixed at different time points during assembly, followed by conductive staining, platinum coating, and visualisation by FEISEM. Changes on the nuclear surface with time are shown. Details of the surface of chromatin and the cytoplasmic face of NPC structure are demonstrated without the need to isolate the structures from the nucleus.


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