Ex vivo time-lapse confocal imaging of the mouse embryo aorta

Jean Boisset, C Andrieu-Soler, Gert van Cappellen, Thomas Clapes, Catherine Robin

Research output: Contribution to journalArticleAcademicpeer-review

24 Citations (Scopus)

Abstract

Time-lapse confocal microscopy of mouse embryo slices was developed to access and image the living aorta. In this paper, we explain how to label all hematopoietic and endothelial cells inside the intact mouse aorta with fluorescent directly labeled antibodies. Then we describe the technique to cut nonfixed labeled embryos into thick slices that are further imaged by time-lapse confocal imaging. This approach allows direct observation of the dynamic cell behavior in the living aorta, which was previously inaccessible because of its location deep inside the opaque mouse embryo. In particular, this approach is sensitive enough to allow the experimenter to witness the transition from endothelial cells into hematopoietic stem/progenitor cells in the aorta, the first site of hematopoietic stem cell generation during development. The protocol can be applied to observe other embryonic sites throughout mouse development. A complete experiment requires similar to 2 d of practical work.
Original languageUndefined/Unknown
Pages (from-to)1792-1805
Number of pages14
JournalNature Protocols
Volume6
Issue number11
DOIs
Publication statusPublished - 2011

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