The lineaging of fluorescently-labeled Caenorhabditis elegans embryos with StarryNite and AceTree
| Autor: | John I. Murray, Robert H. Waterston, Thomas J. Boyle, Zhirong Bao |
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| Rok vydání: | 2006 |
| Předmět: |
Embryo
Nonmammalian Lineage (genetic) ved/biology Gene Expression Profiling Green Fluorescent Proteins ved/biology.organism_classification_rank.species Embryo Biology biology.organism_classification Embryonic stem cell Fusion protein General Biochemistry Genetics and Molecular Biology Cell biology Green fluorescent protein Gene expression profiling Phenotype Image Processing Computer-Assisted Animals Cell Lineage Caenorhabditis elegans Model organism Software |
| Zdroj: | Nature Protocols. 1:1468-1476 |
| ISSN: | 1750-2799 1754-2189 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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