title: Long-term time lapse imaging of mouse cochlear explants.
abstract: Here we present a method for long-term time-lapse imaging of live embryonic mouse cochlear explants. The developmental program responsible for building the highly ordered, complex structure of the mammalian cochlea proceeds for around ten days. In order to study changes in gene expression over this period and their response to pharmaceutical or genetic manipulation, long-term imaging is necessary. Previously, live imaging has typically been limited by the viability of explanted tissue in a humidified chamber atop a standard microscope. Difficulty in maintaining optimal conditions for culture growth with regard to humidity and temperature has placed limits on the length of imaging experiments. A microscope integrated into a modified tissue culture incubator provides an excellent environment for long term-live imaging. In this method we demonstrate how to establish embryonic mouse cochlear explants and how to use an incubator microscope to conduct time lapse imaging using both bright field and fluorescent microscopy to examine the behavior of a typical embryonic day (E) 13 cochlear explant and Sox2, a marker of the prosensory cells of the cochlea, over 5 days.
authors.affiliation: Biological Sciences Platform, Sunnybrook Research Institute; jfm30@sri.utoronto.ca.
authors.forename: Joanna F
authors.initials: JF
authors.lastname: Mulvaney
authors.affiliation: Biological Sciences Platform, Sunnybrook Research Institute; Department of Otolaryngology - Head and Neck Surgery, University of Toronto; Department of Laboratory Medicine and Pathobiology, University of Toronto.
authors.forename: Alain
authors.initials: A
authors.lastname: Dabdoub
mesh_terms: D000818:Animals; D003051:Cochlea / Q000737:chemistry / Q000196:embryology* / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D055748:SOXB1 Transcription Factors / Q000032:analysis / Q000378:metabolism; D059008:Time-Lapse Imaging / Q000379:methods*; D046509:Tissue Culture Techniques
chemical_list: D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse