Document Detail

Preparation of cultured and isolated cells for X-ray microanalysis.
MedLine Citation:
PMID:  3201205     Owner:  NLM     Status:  MEDLINE    
Various electron microscopical preparation techniques are compared with regard to the preservation of the intracellular element distribution as determined by X-ray microanalysis in scanning and scanning transmission electron microscopy. By use of chemical agents for fixation and dehydration ions are redistributed and washed out. This is also true for freeze-substitution. Whole cells are prepared by cryofixation followed by freeze-drying. Interference of intracellular measurements by extracellular elements can be avoided by appropriate washing the cells before cryofixation. The washing medium has to be carefully selected in order to avoid distortions of the original intracellular element content. These problems are circumvented by the preparation of freeze-dried cryosections from cryofixed cells. This is demonstrated by data of the intracellular elemental composition in cultured cells (fibroblasts, Staphylococcus aureus bacteria) and in cells isolated from rat tissue (kidney papillary collecting duct and liver). Cryofixation of a single cell in a defined functional state is illustrated by results obtained from streaming Amoeba proteus cells, cryofixed under light microscopical control. The main conclusion is that X-ray microanalysis of cells in functional states requires cryofixation and cryopreparation techniques which have to be adapted to the particular cell biological problem to be investigated.
K Zierold; D Schäfer
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Scanning microscopy     Volume:  2     ISSN:  0891-7035     ISO Abbreviation:  Scanning Microsc.     Publication Date:  1988 Sep 
Date Detail:
Created Date:  1989-01-19     Completed Date:  1989-01-19     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  8704616     Medline TA:  Scanning Microsc     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1775-90     Citation Subset:  IM    
Max-Planck-Institut für Systemphysiologie, Dortmund, FRG.
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MeSH Terms
Amoeba / analysis*
Cells, Cultured
Electron Probe Microanalysis / methods*
Fibroblasts / analysis*
Freeze Drying
Kidney / analysis,  cytology
Liver / analysis,  cytology
Microscopy, Electron, Scanning
Preservation, Biological
Staphylococcus aureus / analysis*

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