Live imaging and analysis of postnatal mouse retinal development

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dc.contributor.authorNickerson, Philip EB
dc.contributor.authorRonellenfitch, Kara M
dc.contributor.authorCsuzdi, Nicklaus F
dc.contributor.authorBoyd, Jamie D
dc.contributor.authorHoward, Perry L
dc.contributor.authorDelaney, Kerry R
dc.contributor.authorChow, Robert L
dc.date.accessioned2015-03-23T20:50:32Z
dc.date.available2015-03-23T20:50:32Z
dc.date.copyright2013en_US
dc.date.issued2013-06-10
dc.descriptionBioMed Centralen_US
dc.description.abstractBackground: The explanted, developing rodent retina provides an efficient and accessible preparation for use in gene transfer and pharmacological experimentation. Many of the features of normal development are retained in the explanted retina, including retinal progenitor cell proliferation, heterochronic cell production, interkinetic nuclear migration, and connectivity. To date, live imaging in the developing retina has been reported in nonmammalian and mammalian whole-mount samples. An integrated approach to rodent retinal culture/transfection, live imaging, cell tracking, and analysis in structurally intact explants greatly improves our ability to assess the kinetics of cell production. Results: In this report, we describe the assembly and maintenance of an in vitro, CO2-independent, live mouse retinal preparation that is accessible by both upright and inverted, 2-photon or confocal microscopes. The optics of this preparation permit high-quality and multi-channel imaging of retinal cells expressing fluorescent reporters for up to 48h. Tracking of interkinetic nuclear migration within individual cells, and changes in retinal progenitor cell morphology are described. Follow-up, hierarchical cluster screening revealed that several different dependent variable measures can be used to identify and group movement kinetics in experimental and control samples. Conclusions: Collectively, these methods provide a robust approach to assay multiple features of rodent retinal development using live imaging.en_US
dc.description.reviewstatusRevieweden_US
dc.description.scholarlevelFacultyen_US
dc.description.sponsorshipThis work was supported by an operating grant from the Canadian Institutes for Health Research operating grant (RLC). RLC is supported by a Tier 2 Canada Research Chair.en_US
dc.identifier.citationNickerson et al.: Live imaging and analysis of postnatal mouse retinal development. BMC Developmental Biology 2013 13:24.en_US
dc.identifier.urihttp://www.biomedcentral.com/1471-213X/13/24
dc.identifier.urihttp://dx.doi.org/10.1186/1471-213X-13-24
dc.identifier.urihttp://hdl.handle.net/1828/5921
dc.language.isoenen_US
dc.publisherBMC Developmental Biologyen_US
dc.rights.tempAttribution-NonCommercial-NoDerivs 2.5 Canada*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/2.5/ca/*
dc.subjectPostnatal retina
dc.subjectin vitro electroporation
dc.subjectex vivo culture
dc.subjectLive 2-photon microscopy
dc.subjectHierarchical cluster analysis
dc.subject.departmentDepartment of Biology
dc.subject.departmentDepartment of Biochemistry and Microbiology
dc.titleLive imaging and analysis of postnatal mouse retinal developmenten_US
dc.typeArticleen_US

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