Cite as: Cold Spring Harb. Protoc.; 2007; doi:10.1101/pdb.prot4915

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Fate-Mapping Technique: Using Carbocyanine Dyes for Vital Labeling of Cells in Gastrula-Stage Mouse Embryos Cultured In Vitro

Heidi Bildsoe, Vanessa Franklin, and Patrick P.L. Tam1

Children’s Medical Research Institute, University of Sydney, Wentworthville, New South Wales 2145, Australia

1Corresponding author (ptam{at}cmri.usyd.edu.au)


INTRODUCTION

The allocation of different progenitor populations to embryonic structures can be visualized by tracking the distribution of cells to specific tissues in the live embryo. A critical prerequisite for cell tracking is to identify unambiguously the progenitors and their descendants during morphogenesis. This can be achieved by using molecular markers that are expressed from transgenes integrated into the genome or as episomal DNA constructs, or by tagging the cells with exogenous markers that are incorporated into the cell membrane or cytoplasmic components of the cells. These labels can be introduced by dye-labeling the membrane, injecting marker enzyme into the cytoplasm, or integrating reporter constructs by transfection or electroporation. This protocol describes how to label cells in the endoderm (which, at this stage of development, is the superficial tissue layer) of live mouse embryos at 7.0-7.5 days post-coitum (dpc), using two carbocyanine dyes (DiI and DiO).


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