Protocol abstract
Nature Protocols 2, - 3074 - 3080 (2007)
Published online: 29 November 2007 | doi:10.1038/nprot.2007.417
Subject Categories: Cell and developmental biology | Imaging | Model organisms
Live imaging of epidermal morphogenesis during the development of the adult abdominal epidermis of Drosophila
Nikolay Ninov1 & Enrique Martín-Blanco1
Abstract
During larval stages of Drosophila development, the abdominal epidermis is composed of histoblasts (adult precursors) and larval epidermal cells (LECs). During metamorphosis, histoblasts proliferate and colonize the territories occupied by the LECs, which die and become engulfed by macrophages. This morphogenetic process is an excellent model for in vivo analysis of epithelial migration, cell division, cell death, patterning and differentiation. Here, we describe a protocol for time-lapse recording of the developing epidermis during metamorphosis. The protocol describes the removal of the pupal case (which acts as an opaque barrier to effective imaging) and mounting and imaging of specimens of different stages so that normal developmental processes are preserved. This method enables high-resolution studies over long time periods using fluorescent markers and confocal microscopy. The protocol requires 1 h for pupal dissection and mounting and, depending on the stages and genotypes to be analyzed, several more hours for preprocessing and aging and developmental staging of flies and pupae.
- Instituto de Biología Molecular de Barcelona (Consejo Superior de Investigaciones Científicas), Parc Cientific de Barcelona, Josep Samitier 1-5, Barcelona 08028, Spain.
Correspondence to: Enrique Martín-Blanco1 e-mail: embbmc@cid.csic.es
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A-Z product listing
- 1"
36 Yards, Clear(3M) - 3" Core(3M)
- Double-sided sticky tape(3M)
- Double-Sided Tape(3M)
- Stereoscopic epifluorescence microscope (Leica MZ16F(Leica)
- Two pairs of sharp forceps Dumont No. 5 stainless steel(Dumont)
