High pressure freezing and freeze substitution of Schizosaccharomyces pombe and Saccharomyces cerevisiae for TEM.

2.50
Hdl Handle:
http://hdl.handle.net/10541/55804
Title:
High pressure freezing and freeze substitution of Schizosaccharomyces pombe and Saccharomyces cerevisiae for TEM.
Authors:
Murray, Stephen M
Abstract:
The use of standard room temperature chemical fixation protocols for the ultrastructural preservation of yeast and subsequent observation under the electron microscope is fraught with difficulties. Many protocols require the use of enzymatic digestion of the cell wall in order to facilitate the entry of fixatives into the cell interior. Others rely on the use of permanganate-based fixative solutions, which whilst enabling overall preservation of the cell, does require multiple centrifugation, washing, and resuspension steps. This often results in the significant loss of sample volume whilst the use of permanganate can cause extraction of cytoplasmic components. The use of low temperature techniques and in particular high pressure freezing (HPF) and freeze substitution (FS) overcomes many of these problems. With the recent advances in cryotechnologies and in particular the development of commercially available equipment such as the high pressure freezer, the level of ultrastructural preservation attainable in electron microscopy has increased markedly. It is now possible to capture dynamic time sensitive events and to place them in their ultrastructural context with a level of resolution which at the present time can only be achieved with electron microscopy.
Affiliation:
TEM Service Facility, Paterson Institute for Cancer Research, University of Manchester, Manchester, United Kingdom.
Citation:
High pressure freezing and freeze substitution of Schizosaccharomyces pombe and Saccharomyces cerevisiae for TEM. 2008, 88:3-17 Methods Cell Biol.
Journal:
Methods in Cell Biology
Issue Date:
2008
URI:
http://hdl.handle.net/10541/55804
DOI:
10.1016/S0091-679X(08)00401-9
PubMed ID:
18617025
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.date.accessioned2009-03-16T18:08:54Z-
dc.date.available2009-03-16T18:08:54Z-
dc.date.issued2008-
dc.identifier.citationHigh pressure freezing and freeze substitution of Schizosaccharomyces pombe and Saccharomyces cerevisiae for TEM. 2008, 88:3-17 Methods Cell Biol.en
dc.identifier.issn0091-679X-
dc.identifier.pmid18617025-
dc.identifier.doi10.1016/S0091-679X(08)00401-9-
dc.identifier.urihttp://hdl.handle.net/10541/55804-
dc.description.abstractThe use of standard room temperature chemical fixation protocols for the ultrastructural preservation of yeast and subsequent observation under the electron microscope is fraught with difficulties. Many protocols require the use of enzymatic digestion of the cell wall in order to facilitate the entry of fixatives into the cell interior. Others rely on the use of permanganate-based fixative solutions, which whilst enabling overall preservation of the cell, does require multiple centrifugation, washing, and resuspension steps. This often results in the significant loss of sample volume whilst the use of permanganate can cause extraction of cytoplasmic components. The use of low temperature techniques and in particular high pressure freezing (HPF) and freeze substitution (FS) overcomes many of these problems. With the recent advances in cryotechnologies and in particular the development of commercially available equipment such as the high pressure freezer, the level of ultrastructural preservation attainable in electron microscopy has increased markedly. It is now possible to capture dynamic time sensitive events and to place them in their ultrastructural context with a level of resolution which at the present time can only be achieved with electron microscopy.en
dc.language.isoenen
dc.subjectSchizosaccharomyces Pombeen
dc.subjectSaccharomyces Cerevisiaeen
dc.subjectTEMen
dc.subjectHigh Pressure Freezingen
dc.subject.meshAtmospheric Pressure-
dc.subject.meshCryopreservation-
dc.subject.meshFreeze Substitution-
dc.subject.meshFreezing-
dc.subject.meshMicroscopy, Electron, Transmission-
dc.subject.meshSaccharomyces Cerevisiae-
dc.subject.meshSchizosaccharomyces-
dc.titleHigh pressure freezing and freeze substitution of Schizosaccharomyces pombe and Saccharomyces cerevisiae for TEM.en
dc.typeBook chapteren
dc.contributor.departmentTEM Service Facility, Paterson Institute for Cancer Research, University of Manchester, Manchester, United Kingdom.en
dc.identifier.journalMethods in Cell Biologyen
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