Document Detail


High-pressure freezing in the study of animal pathogens.
MedLine Citation:
PMID:  14516363     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
High-pressure freezing is applicable to both morphological and immunocytochemical studies. We are investigating the morphogenesis of foot-and-mouth disease virus and African swine fever virus by the use of high-pressure freezing of infected cells. Foot-and-mouth disease virus particles are not detected in sections of conventionally immersion-fixed infected cells, but when the cells are prepared by high-pressure freezing, newly formed virions are readily seen throughout the cell. We report two methods for high-pressure freezing of virally infected cells: first, two sapphire discs frozen 'face to face' with a narrow spacer to prevent cell damage and, second, a fibrous filter substrate that can be easily cut into discs to fit into the freezing planchettes. Cells readily adhere to the fibres in vitro, and the complete disc can be rapidly transferred to the planchettes for freezing. Immunolabelling studies of the microneme proteins of the parasite Eimeria tenella indicate that high-pressure freezing followed by freeze-substitution in acetone with uranyl acetate allows high-sensitivity immunolabelling for these proteins.
Authors:
P Monaghan; H Cook; P Hawes; J Simpson; F Tomley
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of microscopy     Volume:  212     ISSN:  0022-2720     ISO Abbreviation:  J Microsc     Publication Date:  2003 Oct 
Date Detail:
Created Date:  2003-09-30     Completed Date:  2003-11-12     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0204522     Medline TA:  J Microsc     Country:  England    
Other Details:
Languages:  eng     Pagination:  62-70     Citation Subset:  IM    
Affiliation:
Institute for Animal Health, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK. paul.monaghan@bbsrc.ac.uk
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MeSH Terms
Descriptor/Qualifier:
African Swine Fever Virus / pathogenicity,  ultrastructure*
Animals
Cell Line
Cricetinae
Cryopreservation / instrumentation*,  methods*
Eimeria tenella / pathogenicity,  ultrastructure*
Foot-and-Mouth Disease Virus / pathogenicity,  ultrastructure*
Freeze Substitution
Immunohistochemistry
Microscopy, Electron, Scanning
Pressure

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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