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

被引：79
|
作者：
Murray, John Isaac [1 ]
Bao, Zhirong [1 ]
Boyle, Thomas J. [1 ]
Waterston, Robert H. [1 ]
机构：
[1] Univ Washington, Dept Genome Sci, Seattle, WA 98195 USA
来源：
NATURE PROTOCOLS | 2006年 / 1卷 / 03期
基金：
美国国家卫生研究院;
关键词：
D O I：
10.1038/nprot.2006.222
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
引用
收藏
页码：1468 / 1476
页数：9
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