Document Detail

Imaging surface and submembranous structures with the atomic force microscope: a study on living cancer cells, fibroblasts and macrophages.
MedLine Citation:
PMID:  9674158     Owner:  NLM     Status:  MEDLINE    
Atomic force microscopy (AFM) has been used to image a wide variety of cells. Fixed and dried-coated, wet-fixed or living cells were investigated. The major advantage of AFM over SEM is the avoidance of vacuum and electrons, whereas imaging can be done at environmental pressure and in aqueous conditions. Evidence of the successful application of AFM in biological imaging is provided by comparing results of AFM with SEM and/or TEM. In this study, we investigated surface and submembranous structures of living and glutaraldehyde-fixed colon carcinoma cells, skin fibroblasts and liver macrophages by AFM. Special attention was paid to the correct conditions for the acquisition of images of the surface of these cells, because quality SEM examinations have already been abundantly presented. AFM imaging of living cells revealed specific structures, such as the cytoskeleton, which were not observed by SEM. Membrane structures, such as ruffles, lamellipodia, microspikes and microvilli, could only clearly be observed after fixing the cells with 0.1% glutaraldehyde. AFM images of living cells were comparable to SEM images of fixed, dried and coated cells, but contained a number of artefacts due to tip-sample interaction. In addition, AFM imaging allowed the visualization of cytoplasmic submembranous structures without the necessity for further preparative steps, allowing us: (i) to follow cytoskeletal changes in fibroblasts under the influence of the microfilament disrupting agent latrunculin A; (ii) to study particle phagocytosis in macrophages. Therefore, in spite of the slow image acquisition of the AFM, the instrument can be used for high-resolution real-time studies of dynamic changes in submembranous structures.
F Braet; C Seynaeve; R De Zanger; E Wisse
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of microscopy     Volume:  190     ISSN:  0022-2720     ISO Abbreviation:  J Microsc     Publication Date:  1998 Jun 
Date Detail:
Created Date:  1998-08-18     Completed Date:  1998-08-18     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0204522     Medline TA:  J Microsc     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  328-38     Citation Subset:  IM    
Laboratory for Cell Biology and Histology, Free University of Brussels, Jette, Belgium.
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MeSH Terms
Adenocarcinoma / ultrastructure*
Cells, Cultured
Colonic Neoplasms / ultrastructure*
Fibroblasts / physiology,  ultrastructure*
Liver / cytology,  ultrastructure
Macrophages / physiology,  ultrastructure*
Microscopy, Atomic Force / methods*
Microscopy, Electron / methods*
Rats, Wistar
Skin / cytology,  ultrastructure
Tumor Cells, Cultured

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