Document Detail

Rapidly excised and cryofixed rat tissue.
MedLine Citation:
PMID:  20869536     Owner:  NLM     Status:  MEDLINE    
All preparation efforts of biological samples in electron microscopy are focused to preserve structures as close as possible to the native state. To achieve this goal with tissues, it is of advantage to have a very short time between excision and fixation. The most common approach is chemical fixation: cross-linking of the tissue samples with aldehydes followed by postfixation with osmium tetroxide. Here, the fastest approach for tissue samples is perfusion. However, the diffusion of the fixation solution from blood vessels into the depth of the tissue is still slow and does not allow an overall instant fixation of a single cell. As a result, osmotic effects become evident (swelling or shrinkage of cell organelles). Another possibility is to take a tissue sample from the experimental animal. Excision of tissue can last quite some time, which results in even more pronounced autolytic induced osmotic effects. Furthermore, the animal does not survive the procedure in most cases. Alternatively, microbiopsies are an elegant technique to rapidly excise small quantities of tissue. Some tissues, such as liver and muscle, may be obtained using a non-lethal approach. To avoid the artifacts introduced by chemical fixation, high-pressure freezing of microbiopsies (brain, liver, kidney, and muscle) is a powerful alternative to chemical fixation. Here, we describe the microbiopsy method, and high-pressure freezing/freeze-substitution (HPF/FS) as a follow-up procedure. Cryosectioning of high-pressure frozen samples is optimally preserving the ultrastructure; however, it is not considered to be a routine approach yet.
Dimitri Vanhecke; Werner Graber; Daniel Studer
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Methods in cell biology     Volume:  96     ISSN:  0091-679X     ISO Abbreviation:  Methods Cell Biol.     Publication Date:  2010  
Date Detail:
Created Date:  2010-09-27     Completed Date:  2011-01-20     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0373334     Medline TA:  Methods Cell Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  513-27     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier Inc. All rights reserved.
Institute of Anatomy, University of Berne, CH-3000 Bern 9, Switzerland.
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MeSH Terms
Biopsy* / instrumentation,  methods
Brain / ultrastructure
Cryopreservation / methods*
Freeze Substitution / methods
Histocytological Preparation Techniques / methods*
Liver / ultrastructure
Microscopy, Electron / methods*
Muscles / ultrastructure
Rats / anatomy & histology*
Rats, Wistar

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