Document Detail


Human chromosomes: evaluation of processing techniques for scanning electron microscopy.
MedLine Citation:
PMID:  8316815     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Methods for scanning electron microscopy (SEM) of chromosomes have been developed in the last two decades. Technical limitations in the study of human chromosomes, however, have hindered the routine use of SEM in clinical and experimental human cytogenetics. We compared different methodologies, including metal impregnation, air drying and specimen coating. SEM preparation of human chromosomes in which osmium impregnation is mediated by tannic acid, yielded more reproducible results when compared with osmium impregnation protocols previously described. The level of osmium impregnation was systematically evaluated by imaging chromosomes in the backscattering mode. Critical point drying and a light gold-palladium coating were essential for appropriate secondary electron imaging of chromosomes. With this method, and in a preliminary quantitative analysis, we show that our SEM technique is more sensitive than light microscopy for the detection of aphidicolin-induced fragile sites. This technical approach is useful for chromosomal studies requiring resolution higher than that obtained by light microscopy. Also, it allows the use of clinical and archival chromosomal samples prepared by routine cytogenetic techniques.
Authors:
O H Sanchez-Sweatman; E P de Harven; I D Dubé
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Scanning microscopy     Volume:  7     ISSN:  0891-7035     ISO Abbreviation:  Scanning Microsc.     Publication Date:  1993 Mar 
Date Detail:
Created Date:  1993-07-28     Completed Date:  1993-07-28     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8704616     Medline TA:  Scanning Microsc     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  97-104; discussion 105-6     Citation Subset:  IM    
Affiliation:
Department of Pathology, University of Toronto, Canada.
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MeSH Terms
Descriptor/Qualifier:
Adult
Chromosome Fragile Sites
Chromosome Fragility
Chromosomes, Human / ultrastructure*
Desiccation
Histocytological Preparation Techniques*
Humans
Hydrolyzable Tannins
Male
Microscopy, Electron, Scanning / methods*
Osmium Tetroxide
Chemical
Reg. No./Substance:
0/Hydrolyzable Tannins; 20816-12-0/Osmium Tetroxide

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


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