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

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

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)1502-1510
Number of pages9
JournalNature Protocols
Volume7
Issue number8
DOIs
StatePublished - Aug 2012

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Larva
Animals
Fluorescence
Throughput
Sorting
Caenorhabditis elegans
Homozygote
Flow Cytometry
Genotype
Cells
Equipment and Supplies
Population

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

High-throughput fluorescence-based isolation of live C. elegans larvae. / Fernandez, Anita G.; Bargmann, Bastiaan O R; Mis, Emily K.; Edgley, Mark L.; Birnbaum, Kenneth D.; Piano, Fabio.

In: Nature Protocols, Vol. 7, No. 8, 08.2012, p. 1502-1510.

Research output: Contribution to journalArticle

Fernandez, Anita G. ; Bargmann, Bastiaan O R ; Mis, Emily K. ; Edgley, Mark L. ; Birnbaum, Kenneth D. ; Piano, Fabio. / High-throughput fluorescence-based isolation of live C. elegans larvae. In: Nature Protocols. 2012 ; Vol. 7, No. 8. pp. 1502-1510.
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