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dc.contributor.authorKiseleva, Elena
dc.contributor.authorAllen, Terence D
dc.contributor.authorRutherford, Sandra A
dc.contributor.authorMurray, Stephen M
dc.contributor.authorMorozova, Ksenia N
dc.contributor.authorGardiner, Fiona
dc.contributor.authorGoldberg, Martin W
dc.contributor.authorDrummond, Sheona P
dc.date.accessioned2009-06-15T11:37:04Z
dc.date.available2009-06-15T11:37:04Z
dc.date.issued2007
dc.identifier.citationA protocol for isolation and visualization of yeast nuclei by scanning electron microscopy (SEM). 2007, 2 (8):1943-53 Nat Protocen
dc.identifier.issn1750-2799
dc.identifier.pmid17703206
dc.identifier.doi10.1038/nprot.2007.251
dc.identifier.urihttp://hdl.handle.net/10541/70458
dc.description.abstractThis protocol details methods for the isolation of yeast nuclei from budding yeast (Saccharomyces cerevisiae) and fission yeast (Schizosaccharomyces pombe), immuno-gold labeling of proteins and visualization by field emission scanning electron microscopy (FESEM). This involves the removal of the yeast cell wall and isolation of the nucleus from within, followed by subsequent processing for high-resolution microscopy. The nuclear isolation step can be performed in two ways: enzymatic treatment of yeast cells to rupture the cell wall and generate spheroplasts (cells that have partially lost their cell wall and their characteristic shape), followed by isolation of the nuclei by centrifugation or homogenization; and whole cell freezing followed by manual cell rupture and centrifugation. This protocol has been optimized for the visualization of the yeast nuclear envelope (NE), nuclear pore complexes (NPCs) and associated cyto-skeletal structures. Samples once processed for FESEM can be stored under vacuum for weeks, allowing considerable time for image acquisition.
dc.language.isoenen
dc.subject.meshCell Culture Techniques
dc.subject.meshCell Fractionation
dc.subject.meshCell Nucleus
dc.subject.meshCytoskeleton
dc.subject.meshImmunohistochemistry
dc.subject.meshMicroscopy, Electron, Scanning
dc.subject.meshNuclear Envelope
dc.subject.meshNuclear Pore
dc.subject.meshSaccharomyces cerevisiae
dc.subject.meshSchizosaccharomyces
dc.titleA protocol for isolation and visualization of yeast nuclei by scanning electron microscopy (SEM).en
dc.typeArticleen
dc.contributor.departmentInstitute of Cytology and Genetics, Russian Academy of Science, Novosibirsk, Russia. elka@bionet.nsu.ruen
dc.identifier.journalNature Protocolsen
html.description.abstractThis protocol details methods for the isolation of yeast nuclei from budding yeast (Saccharomyces cerevisiae) and fission yeast (Schizosaccharomyces pombe), immuno-gold labeling of proteins and visualization by field emission scanning electron microscopy (FESEM). This involves the removal of the yeast cell wall and isolation of the nucleus from within, followed by subsequent processing for high-resolution microscopy. The nuclear isolation step can be performed in two ways: enzymatic treatment of yeast cells to rupture the cell wall and generate spheroplasts (cells that have partially lost their cell wall and their characteristic shape), followed by isolation of the nuclei by centrifugation or homogenization; and whole cell freezing followed by manual cell rupture and centrifugation. This protocol has been optimized for the visualization of the yeast nuclear envelope (NE), nuclear pore complexes (NPCs) and associated cyto-skeletal structures. Samples once processed for FESEM can be stored under vacuum for weeks, allowing considerable time for image acquisition.


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