Document Detail


Microcarriers for high-pressure freezing and cryosectioning of adherent cells.
MedLine Citation:
PMID:  18445159     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A method is described employing microcarrier spheres of cross-linked dextran for obtaining ultra- and semithin vitreous sections from high-pressure frozen anchorage-dependent (mammalian) cells. Avoiding trypsination or scraping cells off from the culture surface, the presented approach allows for cryoimmobilization, cryosectioning and cryoelectron microscopy/tomography of frozen-hydrated cells in an unperturbed manner which is important to preserve the native state of, for instance, the cytoskeleton. Furthermore, our studies on the 'life cycle' of Herpes simplex virus in Vero cells demonstrate that cell monolayers on microcarrier beads are well suited for fluorescence microscopic characterization of the sample prior to high-pressure freezing.
Authors:
C Hagen; K Grünewald
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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:  230     ISSN:  1365-2818     ISO Abbreviation:  J Microsc     Publication Date:  2008 May 
Date Detail:
Created Date:  2008-04-30     Completed Date:  2008-06-26     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0204522     Medline TA:  J Microsc     Country:  England    
Other Details:
Languages:  eng     Pagination:  288-96     Citation Subset:  IM    
Affiliation:
Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, D-82152 Martinsried, Germany. hagen@biochem.mpg.de
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Adhesion
Cell Culture Techniques*
Cercopithecus aethiops
Cryoelectron Microscopy / methods*
Cryopreservation
Cryoultramicrotomy / instrumentation*
Dextrans*
Freezing*
Frozen Sections
Herpesvirus 1, Human / physiology,  ultrastructure
Microscopy, Fluorescence / methods*
Microspheres*
Vero Cells / ultrastructure,  virology
Chemical
Reg. No./Substance:
79920-52-8/Cytodex; 9004-54-0/Dextrans

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


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