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Nat Protoc. 2006;1(3):1468-76.

The lineaging of fluorescently-labeled Caenorhabditis elegans embryos with StarryNite and AceTree.

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  • 1Department of Genome Sciences, University of Washington Box 357730, Seattle, Washington 98195, USA.

Abstract

Lineage analysis of Caenorhabditis elegans is a powerful tool for characterizing developmental phenotypes and embryonic gene-expression patterns. We present a detailed protocol for the lineaging of embryos by computational analysis of 4D images of embryos that ubiquitously express histone-GFP (green fluorescent protein) fusion proteins through the 350 cell stage followed by manual editing. We describe how to optimize imaging settings for this purpose, the use of the lineage-extraction software, StarryNite, and the lineage-editing software, AceTree. In addition, we describe a useful polymer bead mounting technique for C. elegans embryos that has several advantages compared with the standard agar pad mounting technique. The protocol requires about 1 h of user time spread over 2 days to generate the raw lineage, and an additional 2 or 4 h to edit the lineage to the 194- or 350-cell stage, respectively.

[PubMed - indexed for MEDLINE]
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