Document Detail


Improved visualization and quantitative analysis of drug effects using micropatterned cells.
MedLine Citation:
PMID:  21189468     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To date, most HCA (High Content Analysis) studies are carried out with adherent cell lines grown on a homogenous substrate in tissue-culture treated micro-plates. Under these conditions, cells spread and divide in all directions resulting in an inherent variability in cell shape, morphology and behavior. The high cell-to-cell variance of the overall population impedes the success of HCA, especially for drug development. The ability of micropatterns to normalize the shape and internal polarity of every individual cell provides a tremendous opportunity for solving this critical bottleneck (1-2). To facilitate access and use of the micropatterning technology, CYTOO has developed a range of ready to use micropatterns, available in coverslip and microwell formats. In this video article, we provide detailed protocols of all the procedures from cell seeding on CYTOOchip micropatterns, drug treatment, fixation and staining to automated acquisition, automated image processing and final data analysis. With this example, we illustrate how micropatterns can facilitate cell-based assays. Alterations of the cell cytoskeleton are difficult to quantify in cells cultured on homogenous substrates, but culturing cells on micropatterns results in a reproducible organization of the actin meshwork due to systematic positioning of the cell adhesion contacts in every cell. Such normalization of the intracellular architecture allows quantification of even small effects on the actin cytoskeleton as demonstrated in these set of protocols using blebbistatin, an inhibitor of the actin-myosin interaction.
Authors:
Sébastien Degot; Muriel Auzan; Violaine Chapuis; Anne Béghin; Amélie Chadeyras; Constantin Nelep; Maria Luisa Calvo-Muñoz; Joanne Young; François Chatelain; Alexandra Fuchs
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Publication Detail:
Type:  Journal Article; Video-Audio Media     Date:  2010-12-02
Journal Detail:
Title:  Journal of visualized experiments : JoVE     Volume:  -     ISSN:  1940-087X     ISO Abbreviation:  J Vis Exp     Publication Date:  2010  
Date Detail:
Created Date:  2010-12-29     Completed Date:  2011-01-20     Revised Date:  2013-07-03    
Medline Journal Info:
Nlm Unique ID:  101313252     Medline TA:  J Vis Exp     Country:  United States    
Other Details:
Languages:  eng     Pagination:  -     Citation Subset:  IM    
Affiliation:
CYTOO Cell Architects, Grenoble, France. sdegot@cytoo.com
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MeSH Terms
Descriptor/Qualifier:
Actins / antagonists & inhibitors,  metabolism
Cell Adhesion
Cytological Techniques / methods*
Cytoskeleton / drug effects,  metabolism
Drug Evaluation, Preclinical / methods*
HeLa Cells
Heterocyclic Compounds with 4 or More Rings / pharmacology
Humans
Myosins / antagonists & inhibitors,  metabolism
Staining and Labeling / methods
Chemical
Reg. No./Substance:
0/Actins; 0/Heterocyclic Compounds with 4 or More Rings; 0/blebbistatin; EC 3.6.4.1/Myosins
Comments/Corrections

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


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