Document Detail

Wide confocal cytometry: a new approach to study proteomic and structural changes in the cell nucleus during the cell cycle.
MedLine Citation:
PMID:  17989992     Owner:  NLM     Status:  MEDLINE    
Wide-confocal-cytometry (WCC) is a new method developed in this paper that uses a standard confocal system to gather quantitative information on contents and fine structural details of cells. The system is operated under conditions of non-confocality, in order to capture the maximum amount of light emitted by the specimen (comparable to LSC). After analysis of macromolecule content (DNA, RNA, specific proteins, lipids, etc.), cells can be sampled using conventional confocal microscopy. We analyzed the illumination and acquiring capabilities of WCC. The quantitative power of WCC was validated by analysis of cell cycle stage in Hela cells, looking at DNA content and markers for S phase and mitosis. As an example of the potential of this methodology we have documented changes in cell nucleus during the cell cycle. After mitosis the cell nucleus changes its shape from elongated to ellipsoid and remains constant until G2. This change is associated with nuclear volume increase. As nuclear volume increases, chromatin becomes decondensed in an isometric manner, probably due to the increase in gene expression and factors necessary for RNA metabolism.
Francisco J Iborra; Veronica Buckle
Publication Detail:
Type:  Journal Article     Date:  2007-11-08
Journal Detail:
Title:  Histochemistry and cell biology     Volume:  129     ISSN:  0948-6143     ISO Abbreviation:  Histochem. Cell Biol.     Publication Date:  2008 Jan 
Date Detail:
Created Date:  2007-12-14     Completed Date:  2009-01-29     Revised Date:  2014-02-19    
Medline Journal Info:
Nlm Unique ID:  9506663     Medline TA:  Histochem Cell Biol     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  45-53     Citation Subset:  IM    
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MeSH Terms
Cell Cycle / physiology*
Cell Nucleus*
Cells, Cultured
HeLa Cells
Microscopy, Confocal / methods*
Proteomics / methods*
Grant Support
MC_U137961143//Medical Research Council; MC_U137973816//Medical Research Council

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