A protocol for isolation and visualization of yeast nuclei by scanning electron microscopy.

2.50
Hdl Handle:
http://hdl.handle.net/10541/65434
Title:
A protocol for isolation and visualization of yeast nuclei by scanning electron microscopy.
Authors:
Murray, Stephen M; Kiseleva, Elena
Abstract:
This article describes a protocol that details methods for the isolation of yeast nuclei from budding yeast (Saccharomyces cerevisiae) and fission yeast (Schizosaccharomyces pombe), immunogold labelling 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 is performed by 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 nuclei by centrifugation. This protocol has been optimized for the visualization of the yeast nuclear envelope (NE), nuclear pore complexes (NPCs), and associated cytoskeletal structures. Samples, once processed for FESEM, can be stored under vacuum for weeks, allowing considerable time for image acquisition.
Affiliation:
TEM Service Facility, Paterson Institute for Cancer Research, University of Manchester, Manchester M20 4BX, United Kingdom.
Citation:
A protocol for isolation and visualization of yeast nuclei by scanning electron microscopy. 2008, 88:367-87 Methods Cell Biol.
Journal:
Methods in Cell Biology
Issue Date:
2008
URI:
http://hdl.handle.net/10541/65434
DOI:
10.1016/S0091-679X(08)00419-6
PubMed ID:
18617043
Type:
Book chapter
Language:
en
ISSN:
0091-679X
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorMurray, Stephen M-
dc.contributor.authorKiseleva, Elena-
dc.date.accessioned2009-04-20T09:58:11Z-
dc.date.available2009-04-20T09:58:11Z-
dc.date.issued2008-
dc.identifier.citationA protocol for isolation and visualization of yeast nuclei by scanning electron microscopy. 2008, 88:367-87 Methods Cell Biol.en
dc.identifier.issn0091-679X-
dc.identifier.pmid18617043-
dc.identifier.doi10.1016/S0091-679X(08)00419-6-
dc.identifier.urihttp://hdl.handle.net/10541/65434-
dc.description.abstractThis article describes a protocol that details methods for the isolation of yeast nuclei from budding yeast (Saccharomyces cerevisiae) and fission yeast (Schizosaccharomyces pombe), immunogold labelling 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 is performed by 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 nuclei by centrifugation. This protocol has been optimized for the visualization of the yeast nuclear envelope (NE), nuclear pore complexes (NPCs), and associated cytoskeletal structures. Samples, once processed for FESEM, can be stored under vacuum for weeks, allowing considerable time for image acquisition.en
dc.language.isoenen
dc.subject.meshCell Fractionation-
dc.subject.meshCell Nucleus-
dc.subject.meshCell Proliferation-
dc.subject.meshFixatives-
dc.subject.meshImmunohistochemistry-
dc.subject.meshMicroscopy, Electron, Scanning-
dc.subject.meshModels, Biological-
dc.subject.meshRibosomes-
dc.subject.meshTissue Fixation-
dc.subject.meshYeasts-
dc.titleA protocol for isolation and visualization of yeast nuclei by scanning electron microscopy.en
dc.typeBook chapteren
dc.contributor.departmentTEM Service Facility, Paterson Institute for Cancer Research, University of Manchester, Manchester M20 4BX, United Kingdom.en
dc.identifier.journalMethods in Cell Biologyen

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