Document Detail


Plasma etching and ashing: a technique for demonstrating internal structures of helminths using scanning electron microscopy.
MedLine Citation:
PMID:  16469178     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Plasma etching and ashing for demonstrating the three-dimensional ultrastructure of the internal organs of helminths is described. Adult worms of the cestode Caryophyllaeides fennica were dehydrated through an ethanol series, critical point dried (Polaron E3000) and sputter coated with 60% gold-palladium (Polaron E5100) and glued to a standard scanning electron microscope (SEM) stub positioned as required for ashing. After initial SEM viewing of worm surfaces for orientation, stubs were placed individually in the reactor chamber of a PT7150 plasma etching and ashing machine. Worms were exposed to a radio frequency (RF) potential in a low pressure (0.2 mbar) oxygen atmosphere at room temperature. The oxidation process was controlled by varying the times of exposure to the RF potential between 2 to 30 min, depending on the depth of surface tissue to be removed to expose target organs or tissues. After each exposure the oxidized layer was blown from the surface with compressed air, the specimen sputter-coated, and viewed by SEM. The procedure was repeated as necessary, to progressively expose successive layers. Fine details of organs, cells within, and cell contents were revealed. Ashing has the advantage of providing three dimensional images of the arrangement of organs that are impossible to visualize by any other procedure, for example facilitating testes counts in cestodes. Both freshly-fixed and long-term stored helminths can be ashed. Ashing times to obtain the desired results were determined by trial so that some duplicate material was needed.
Authors:
C J Veltkamp; J C Chubb
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of helminthology     Volume:  80     ISSN:  0022-149X     ISO Abbreviation:  J. Helminthol.     Publication Date:  2006 Mar 
Date Detail:
Created Date:  2006-02-10     Completed Date:  2006-08-21     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  2985115R     Medline TA:  J Helminthol     Country:  England    
Other Details:
Languages:  eng     Pagination:  79-82     Citation Subset:  IM    
Affiliation:
School of Biological Sciences, University of Liverpool, Liverpool L69 7ZB, UK.
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MeSH Terms
Descriptor/Qualifier:
Animals
Caryophyllaceae / ultrastructure*
Freeze Etching / methods
Microscopy, Electron, Scanning / methods*
Plasma

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


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