Development is normal during embryo culture. (A) Wild type embryo collected at E7.5 and cultured in vitro for 24 hours develops with (B-C) normal morphology with a yolk sac. (D) Wild type embryos harvested at E8.5 and (E) cultured in vitro for 24 hours increase in size with normal developmental milestones including observable heartbeat (F-N). Immunocytochemistry of the wild type embryos fixed at E9.5 after 24 hours of in vitro culture. Images show normal expression pattern of ventral cell fate markers and proliferation marker Ki67. Shh and Olig2 (F-H), Pax6 and Ki67 (I-K), Nkx2.2 and Olig2 (L-N). Bar is 1.5 μm (A-C), 1 μm (D-E) and 50 μm (F-N).