A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods

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A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods

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dc.contributor.author Yarrow, Justin C
dc.contributor.author Perlman, Zachary E
dc.contributor.author Westwood, Nicholas J
dc.contributor.author Mitchison, Timothy J.
dc.date.accessioned 2011-03-09T04:09:42Z
dc.date.issued 2004
dc.identifier.citation Yarrow, Justin C, Zachary E Perlman, Nicholas J Westwood, and Timothy J Mitchison. 2004. A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods. BMC Biotechnology 4: 21. en_US
dc.identifier.issn 1472-6750 en_US
dc.identifier.uri http://nrs.harvard.edu/urn-3:HUL.InstRepos:4739592
dc.description.abstract Background: Cell migration is a complex phenomenon that requires the coordination of numerous cellular processes. Investigation of cell migration and its underlying biology is of interest to basic scientists and those in search of therapeutics. Current migration assays for screening small molecules, siRNAs, or other perturbations are difficult to perform in parallel at the scale required to screen large libraries. Results: We have adapted the commonly used scratch wound healing assay of tissue-culture cell monolayers to a 384 well plate format. By mechanically scratching the cell substrate with a pin array, we are able to create characteristically sized wounds in all wells of a 384 well plate. Imaging of the healing wounds with an automated fluorescence microscope allows us to distinguish perturbations that affect cell migration, morphology, and division. Readout requires ~1 hr per plate but is high in information content i.e. high content. We compare readouts using different imaging technologies, automated microscopy, scanners and a fluorescence macroscope, and evaluate the trade-off between information content and data acquisition rate. Conclusions: The adaptation of a wound healing assay to a 384 well format facilitates the study of aspects of cell migration, tissue reorganization, cell division, and other processes that underlie wound healing. This assay allows greater than 10,000 perturbations to be screened per day with a quantitative, high-content readout, and can also be used to characterize small numbers of perturbations in detail. en_US
dc.language.iso en_US en_US
dc.publisher BioMed Central en_US
dc.relation.isversionof doi:10.1186/1472-6750-4-21 en_US
dc.relation.hasversion http://www.ncbi.nlm.nih.gov/pmc/articles/PMC521074/pdf/ en_US
dash.license LAA
dc.subject Endothelial Cells en_US
dc.subject In-Vitro en_US
dc.subject Polarity en_US
dc.subject Overexpression en_US
dc.subject Proliferation en_US
dc.subject Involvement en_US
dc.subject Activation en_US
dc.subject Astrocytes en_US
dc.subject Cancer en_US
dc.title A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods en_US
dc.type Journal Article en_US
dc.description.version Version of Record en_US
dc.relation.journal BMC Biotechnology en_US
dash.depositing.author Mitchison, Timothy J.
dc.date.available 2011-03-09T04:09:42Z
dash.affiliation.other HMS^Systems Biology en_US

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