Live imaging and analysis of postnatal mouse retinal development

BMC Developmental Biology

Robert L Chow⁴  Kerry R Delaney⁴  Perry L Howard³  Jamie D Boyd¹  Nicklaus F Csuzdi²  Kara M Ronellenfitch⁴  Philip EB Nickerson⁵ 

[1] Department of Psychiatry, University of British Columbia, Vancouver, BC, Canada;FRAN, University of Guelph, Guelph, ON, Canada;Department of Biochemistry/Microbiology, University of Victoria, Victoria, BC, Canada;Department of Biology, University of Victoria, Station CSC, PO Box 3020, Victoria, BC V8W 3N5, Canada;Current address: Department of Surgery (Neurosurgery), Dalhousie University, Halifax, NS B3H 3A7, Canada

关键词: Hierarchical cluster analysis;    Live 2-photon microscopy;    ex vivo culture;    in vitro electroporation;    Postnatal retina;   

Others  :  1085714
DOI  :  10.1186/1471-213X-13-24

 received in 2012-12-05, accepted in 2013-05-09,  发布年份 2013

【 摘 要 】

Background

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 non-mammalian 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.

【 授权许可】

   
2013 Nickerson et al.; licensee BioMed Central Ltd.

【 图 表 】

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