Protocols were adapted from what was previously described in Handke et al. (2014) (link) and Poulton et al. (2014) (link). Briefly, wing discs were dissected from third instar larvae at room temperature and placed apical side up onto an oxygen-permeable Lumox dish 50 (Sarstedt) in a base media [Schneider’s media (GIBCO, Grand Island, NY), 5% fly extract (Drosophila Genomics Resource Center), 6.2 μg/ml bovine insulin, and 1% penicillin/streptomycin] supplemented with 200 μg/ml bovine insulin. Spacers of 1 μm in thickness were placed adjacent to the disc and a poly-l -lysine-coated coverslip was placed on top. Coverslips were sealed with Halocarbon oil 700 (Sigma Chemical, St. Louis, MO) around the edges. Discs were allowed to settle for 30 min, after which they were imaged with a 40× water objective for up to 3 hr.
Lumox dish 50
Manufactured by Sarstedt
The Lumox dish 50 is a cell culture dish designed for high-quality cell growth and observation. It features a transparent, gas-permeable membrane that allows efficient gas exchange, supporting optimal cell culture conditions. The dish is made of high-quality polystyrene material and is sterile, providing a reliable and consistent environment for cell-based experiments.
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2 protocols using lumox dish 50
1
Imaging Live Drosophila Wing Discs
2
Gut Dissection and Mounting in LIG
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Two pieces of cover glass with a size of 10 × 22 mm were attached to a lumox® dish 50 (Sarstedt, Cat# 15090935) using LIG, with a gap of ~1 cm between them. Intact guts were dissected in LIB and transferred to a 22 × 22 mm cover glass. Excess LIB was carefully removed with filter paper. A volume of 80 µl LIG at 37 °C was dropped into the 1 cm gap, then the 22 × 22 mm cover glass was quickly placed on the top of the 10 × 22 mm cover glasses to cover the guts with LIG without air bubbles. After the LIG was cooled down and stabilized, the cover glasses were finally sealed with Halocarbon oil 27 (Sigma-Aldrich, Cat# H8773) to prevent evaporation.
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