We describe an automated method to isolate mutant Caenorhabditis elegans that do not appropriately execute cellular differentiation programs. We used a fluorescence-activated sorting mechanism implemented in the COPAS Biosort machine to isolate mutants with subtle alterations in the cellular specificity of GFP expression. This methodology is considerably more efficient than comparable manual screens and enabled us to isolate mutants in which dopamine neurons do not differentiate appropriately.
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We thank A. Kruyer, A. Pretorian, J. Recio and Q. Chen for technical assistance, S. Sarin for calculating the degree of saturation, V. Bertrand and F. Zhang for communicating unpublished results, and S. Mitani (Tokyo Women's Medical University School of Medicine) for providing lin-32 deletion alleles. This work was funded by the US National Institutes of Health (R01NS039996-05; R01NS050266-03), the Howard Hughes Medical Institute, an European Molecular Biology Organization long-term fellowship to M.D. and N.F., and a Marie Curie Outgoing International fellowship to N.F.
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Doitsidou, M., Flames, N., Lee, A. et al. Automated screening for mutants affecting dopaminergic-neuron specification in C. elegans. Nat Methods 5, 869–872 (2008). https://doi.org/10.1038/nmeth.1250
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