Technical Reference #1587

1587.

Active Cell Movement Couped To Positional Induction Are Involved In Lineage Segregation In The Mouse Blastocyst Sigolene M. Meilhac; Richard J. Adams; Samantha A. Morris; Anne Danckaert; Jean-Francois Le Garrec; Magdalena Zernicka-Goetz, University of Cambridge, Developmental Biology, 331(1587), (2009)
Link To Paper

Abstract:
In the mouse blastocyst some cells of the inner cell mass (ICM) develop into primitive endoderm (PE) at thesurface while deeper cells form the epiblast. It remained unclear whether the position of cells determinestheir fate such that gene expressio

Keywords:
mouse blastocyst; cell lineage; cell movement; Gata 6; Wnt; Inner cell mass; primitive endoderm

Materials & Methods:
Embryos were cultured and imaged in 4D in a glass-bottom dish (MatTek) on an inverted epifluorescent Zeiss Axiovert 200M microscope with a 20×/0.75NA objective. Multi-channel (red fluorescence/ green fluorescence/transmission) multi-section images were acquired every 10 or 15 min as before (Bischoff et al. 2008). In twochannel (green/transmission) movies 12 focal planes were acquired every 5 μm with an exposure of 4 ms for transmitted light and 400ms for green fluorescence using a 30% cut neutral density filter from Chroma. In three-channel movies 9 focal planes were acquired every 6–7 μm with an exposure of 4 ms for transmitted light 70 ms for green fluorescence and 50 ms for red fluorescence using a 50% cut neutral density filter from Chroma.

Microscopic Technique
Time Lapse Microscopy

Cell Type(s)
embryos