High-throughput fluorescence-based isolation of live C. elegans larvae View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2012-08

AUTHORS

Anita G Fernandez, Bastiaan O R Bargmann, Emily K Mis, Mark L Edgley, Kenneth D Birnbaum, Fabio Piano

ABSTRACT

For the nematode Caenorhabditis elegans, automated selection of animals of specific genotypes from a mixed pool has become essential for genetic interaction or chemical screens. To date, such selection has been accomplished using specialized instruments. However, access to such dedicated equipment is not common. Here we describe live animal fluorescence-activated cell sorting (laFACS), a protocol for automatic selection of live first larval stage (L1) animals using a standard FACS system. We show that FACS can be used for the precise identification of GFP-expressing and non-GFP-expressing subpopulations and can accomplish high-speed sorting of live animals. We have routinely collected 100,000 or more homozygotes from a mixed starting population within 2 h, and with greater than 99% purity. The sorted animals continue to develop normally, making this protocol ideally suited for the isolation of terminal mutants for use in genetic interaction or chemical genetic screens. More... »

PAGES

1502

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nprot.2012.084

DOI

http://dx.doi.org/10.1038/nprot.2012.084

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1046966255

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/22814389


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