Show simple item record

dc.contributor.authorGe, Jing
dc.contributor.authorPrasongtanakij, Somsak
dc.contributor.authorWood, David K.
dc.contributor.authorWeingeist, David M.
dc.contributor.authorFessler, Jessica
dc.contributor.authorNavasummrit, Panida
dc.contributor.authorRuchirawat, Mathuros
dc.contributor.authorEngelward, Bevin P.
dc.date.accessioned2014-12-29T17:45:22Z
dc.date.available2014-12-29T17:45:22Z
dc.date.issued2014-10
dc.identifier.issn1940-087X
dc.identifier.urihttp://hdl.handle.net/1721.1/92527
dc.description.abstractDNA damaging agents can promote aging, disease and cancer and they are ubiquitous in the environment and produced within human cells as normal cellular metabolites. Ironically, at high doses DNA damaging agents are also used to treat cancer. The ability to quantify DNA damage responses is thus critical in the public health, pharmaceutical and clinical domains. Here, we describe a novel platform that exploits microfabrication techniques to pattern cells in a fixed microarray The ‘CometChip’ is based upon the well-established single cell gel electrophoresis assay (a.k.a. the comet assay), which estimates the level of DNA damage by evaluating the extent of DNA migration through a matrix in an electrical field. The type of damage measured by this assay includes abasic sites, crosslinks, and strand breaks. Instead of being randomly dispersed in agarose in the traditional assay, cells are captured into an agarose microwell array by gravity. The platform also expands from the size of a standard microscope slide to a 96-well format, enabling parallel processing. Here we describe the protocols of using the chip to evaluate DNA damage caused by known genotoxic agents and the cellular repair response followed after exposure. Through the integration of biological and engineering principles, this method potentiates robust and sensitive measurements of DNA damage in human cells and provides the necessary throughput for genotoxicity testing, drug development, epidemiological studies and clinical assays.en_US
dc.description.sponsorshipNational Institute of Environmental Health Sciences (Training Grant in Environmental Toxicology T32-ES007020)en_US
dc.description.sponsorshipMassachusetts Institute of Technology. Center for Environmental Health Sciences (P30-ES002109)en_US
dc.description.sponsorshipNational Institute of Environmental Health Sciences (5-UO1-ES016045)en_US
dc.description.sponsorshipNational Institute of Environmental Health Sciences (1-R21-ES019498)en_US
dc.description.sponsorshipNational Institute of Environmental Health Sciences (R44-ES021116)en_US
dc.language.isoen_US
dc.publisherMyJoVE Corporationen_US
dc.relation.isversionofhttp://dx.doi.org/10.3791/50607en_US
dc.rightsCreative Commons Attributionen_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/en_US
dc.sourceMyJoVE Corporationen_US
dc.titleCometChip: A High-throughput 96-Well Platform for Measuring DNA Damage in Microarrayed Human Cellsen_US
dc.typeArticleen_US
dc.identifier.citationGe, Jing, Somsak Prasongtanakij, David K. Wood, David M. Weingeist, Jessica Fessler, Panida Navasummrit, Mathuros Ruchirawat, and Bevin P. Engelward. “CometChip: A High-Throughput 96-Well Platform for Measuring DNA Damage in Microarrayed Human Cells.” JoVE no. 92 (2014).en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.contributor.mitauthorGe, Jingen_US
dc.contributor.mitauthorEngelward, Bevin P.en_US
dc.contributor.mitauthorWeingeist, David M.en_US
dc.contributor.mitauthorFessler, Jessicaen_US
dc.relation.journalJournal of Visualized Experimentsen_US
dc.eprint.versionFinal published versionen_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsGe, Jing; Prasongtanakij, Somsak; Wood, David K.; Weingeist, David M.; Fessler, Jessica; Navasummrit, Panida; Ruchirawat, Mathuros; Engelward, Bevin P.en_US
mit.licensePUBLISHER_CCen_US
mit.metadata.statusComplete


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record