Document Detail


Preparation of chromosome suspensions for flow cytometry.
MedLine Citation:
PMID:  6201326     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Certain variables in the preparation of chromosome suspensions for flow cytometric analysis have been investigated. The optimal conditions have been determined. The results of this series of experiments have been incorporated to yield a preparation protocol that gives chromosome profiles with a low amount of small particle debris and few chromosome clumps. The method reduces variability that results from sample preparation. Chromosomes are optimally isolated in a hypotonic solution buffered to pH 8.0. MgSO4 and dithiothreitol added to the buffer reduce the number of clumps and small fluorescent particles. The presence of MgSO4 also stabilizes the chromosomes and precludes the need for other stabilizing agents such as propidium iodide. When the swelling buffer developed in this investigation is used, unstained chromosomes are stable for at least 1 wk if stored at 4 degrees C. The preparation procedure can be used with the DNA stains, propidium iodide, Hoechst 33258, and mithramycin. Preliminary experiments show that this procedure can also be used for bivariate analysis of human and Chinese hamster chromosomes. The importance of this improvement for studies on chromosome damage caused by irradiation or mutagens is discussed.
Authors:
G van den Engh; B Trask; S Cram; M Bartholdi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Cytometry     Volume:  5     ISSN:  0196-4763     ISO Abbreviation:  Cytometry     Publication Date:  1984 Mar 
Date Detail:
Created Date:  1984-06-19     Completed Date:  1984-06-19     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  8102328     Medline TA:  Cytometry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  108-17     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Line
Chromosomes / drug effects,  ultrastructure*
Cricetinae
Cricetulus
DNA / analysis*
Demecolcine / pharmacology
Detergents
Dithiothreitol / pharmacology
Fetus
Flow Cytometry / methods
Hydrogen-Ion Concentration
Mercaptoethanol / pharmacology
Mitosis / drug effects
Ribonucleases
Staining and Labeling
Vinblastine / pharmacology
Grant Support
ID/Acronym/Agency:
RR 01315/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Detergents; 3483-12-3/Dithiothreitol; 477-30-5/Demecolcine; 60-24-2/Mercaptoethanol; 865-21-4/Vinblastine; 9007-49-2/DNA; EC 3.1.-/Ribonucleases

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


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