Lumox teflon bottom dish

Eyelids were explanted from mid-to-late E15.5 embryos into warm media (Defined Keratinocyte Serum-free Media supplemented with 600 μm calcium and 5% w/v penicillin-streptomycin, Life Technologies). For live imaging, we used methodology previously described for embryonic skin explants (Li et al., 2011 (link)). Using a small volume of growth factor-reduced matrigel, allowed to polymerize for 25 min at 37°C, we sealed eyes against a Lumox teflon-bottom dish (Sarstedt). Eyelid closure was imaged for periods of 6–16 h in 5% CO2 on a PerkinElmer Volocity spinning disk system equipped with a heated enclosure and gas mixer (Solent) and ×20/0.75 CFI Plan-Apo objective, or a Zeiss LSM 780 system with a stage-top incubator and Plan-Apochromat ×20/0.8 objective.

Midgut slices of 200μm (HH17 Tie1:H2B-YFP quails) were made using Vibratome 800, (McIlwain). Explant live imaging was adapted from (Gros et al., 2010 (link)Gros et al., 2010 (link); Heller et al., 2014 (link)). Briefly, midgut explants were suspended in warm recovery media (F12, 10mM HEPES, 1% Pen-strep; Invitrogen) and sealed against a matTek glass- (Movie S1 and S2) or Lumox Teflon-bottom dish (Sarstedt) (Movie S3) using a small volume of 1% low melting point agarose/F12. Images were captured every 7 minutes for 18 hours on a Zeiss LSM 780/Plan-Apo 20×/0.8 setup in an air- and stage-heated 5% CO₂ (Movie S3) or for periods of 6–10 hours using a Leica SP5/Plan-Apo 20×/0.7 setup in a heated, 5% CO₂ (Movie S1 and S2).