Document Detail


Determination of cell number in monolayer cultures.
MedLine Citation:
PMID:  3812988     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Determining the cytostatic or cytotoxic effects of various conditions on monolayer cells requires techniques that are rapid, reproducible, and able to monitor these effects as a function of time. Methods currently used to monitor cytostasis or cytotoxicity are either static or indirect; that is, they are designed to test effects of various treatments either at single time points or on associated cellular processes, such as membrane integrity. Because of these limitations in extant techniques, we undertook this study to improve methods for the rapid determination of cell number in monolayer cultures. We have arrived at conditions of staining cell nuclei with crystal violet under fixed regimens which allow rapid and reproducible quantification of cell number in cultures grown in 24-well miniwells. Quantification is possible by solubilizing the adsorbed dye into a solution of Triton X-100 and determining optical density (O.D.) using spectrophotometry. The present communication documents that O.D. is linearly related to cell number with a sensitivity of ca. 500 cells and that the technique is applicable to study agents which affect cell proliferation.
Authors:
R J Gillies; N Didier; M Denton
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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:  Analytical biochemistry     Volume:  159     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1986 Nov 
Date Detail:
Created Date:  1987-02-25     Completed Date:  1987-02-25     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  109-13     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Count / methods*
Cell Nucleus
Cells, Cultured
Gentian Violet
Mice
Octoxynol
Polyethylene Glycols
Spectrophotometry
Grant Support
ID/Acronym/Agency:
GM 34656/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Polyethylene Glycols; 548-62-9/Gentian Violet; 9002-93-1/Octoxynol

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


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