Document Detail

A novel technique for observing the internal ultrastructure of human chromosomes with known karyotype.
MedLine Citation:
PMID:  18598571     Owner:  NLM     Status:  MEDLINE    
Observation of the internal ultrastructure of human chromosomes by transmission electron microscopy (TEM) has frequently been attempted in spite of the difficulties in detaching metaphase chromosome spreads from the glass slide for further processing. In this study we have used a method in which metaphase chromosome spreads were prepared on a flexible thermoplastic membrane (ACLAR) film. To assess chromosome identity, a diamidino-phenylindole staining and karyotying was first done using a conventional cytogenetic system. The chromosome spreads were then fixed with 1% osmium tetroxide, stained with freshly prepared 2% tannic acid, dehydrated, and flat-embedded in epoxy resin. The resin sheet was easily detachable and carried whole chromosome spreads. By this method, TEM observation of chromosomes from normal human lymphocytes allowed a thorough examination of the ultrastructure of centromeres, telomeres, fragile sites, and other chromosomal regions. Various ultrastructural patterns including thick electron dense boundaries, less dense internal regions, and extended chromatin loops at the periphery of the chromosomes were discernible. Application of the present method to chromosome research is expected to provide comprehensive information on the internal ultrastructure of different chromosomal regions in relation to function.
Mohammad Ghazizadeh; Yoshihiro Sasaki; Tatsuo Oguro; Shigeru Sato; Seiko Egawa; Kyoko Inoue; Akiko Adachi; Hajime Shimizu; Oichi Kawanami
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada     Volume:  14     ISSN:  1431-9276     ISO Abbreviation:  Microsc. Microanal.     Publication Date:  2008 Aug 
Date Detail:
Created Date:  2008-07-07     Completed Date:  2008-09-11     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9712707     Medline TA:  Microsc Microanal     Country:  United States    
Other Details:
Languages:  eng     Pagination:  357-61     Citation Subset:  IM    
Department of Molecular Pathology, Institute of Gerontology, Nippon Medical School, Kawasaki, Japan.
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MeSH Terms
Centromere / ultrastructure
Chromosomes, Human / ultrastructure*
Lymphocytes / ultrastructure
Microscopy, Electron, Transmission / methods*
Staining and Labeling
Telomere / ultrastructure

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