Automated lineage and expression profiling in live Caenorhabditis elegans embryos

Area of Science:

• Developmental Biology
• Molecular Biology
• Genetics

Background:

• Quantitative measurement of gene expression with cellular resolution is crucial for understanding animal development.
• Fluorescent protein reporters enable live-cell imaging, but identifying cells and comparing expression across organisms remains challenging.

Purpose of the Study:

• To present a protocol for automated lineage analysis to identify and quantify fluorescent reporter gene expression in Caenorhabditis elegans embryos.
• To provide a method for detailed analysis of gene expression dynamics during development.

Main Methods:

• Utilizing the invariant cell division pattern of C. elegans for automated lineage tracing and cell identification.
• Employing fluorescent histone transgenes for lineage mapping and a second fluorescent reporter for gene of interest expression.
• Image acquisition, automated lineage analysis, manual curation, and reporter signal extraction.

Main Results:

• Successful identification of expressing cells and quantification of reporter expression with cellular resolution in C. elegans embryos.
• Demonstrated utility for analyzing reporter expression, cell division timing, and cell position in genetically perturbed embryos.

Conclusions:

• Automated lineage analysis provides a robust scaffold for quantifying gene expression dynamics in C. elegans development.
• This protocol enhances the ability to study gene function and developmental processes at single-cell resolution.