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FACS-based isolation of slowly growing cells: Double encapsulation of yeast in gel microdrops

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Abstract

Isolating hyperproducing cells is important in biotechnology, but these cells usually grow slowly and can be overgrown by poorly producing cells. We describe a new method of isolating slowly growing cells from among rapidly growing cells, which has the potential for automation and high throughput (e.g., 100,000 cells/h). A model system is presented consisting of a mixed population of slowly growing mutant and rapidly growing wild-type yeast, which were encapsulated in double agarose gel microdrops (dGMDs); with most dGMDs initially containing single cells. Double encapsulation locates parent cells near dGMD centers, making microcolony measurement more accurate. After a 15-h incubation, fluorescent activated cell sorting was used to analyze and sort dGMDs with small microcolonies (slow growers) from dGMDs with large microcolonies (rapid growers). Successful isolation of slow growers from a mixed population of predominantly rapidly growing Saccharomyces cerevisiae cells was achieved.

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Author notes

  1. James C. Weaver: email: jim@geldrop.mit.edu

Authors and Affiliations

  1. Harvard-M.I.T. Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, 02139

    Elizabeth A. Gift & James C. Weaver

  2. Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 02139

    Hyun J. Park & Arnold L. Demain

  3. Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139

    Glenn A. Paradis

Authors
  1. Elizabeth A. Gift
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  2. Hyun J. Park
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  3. Glenn A. Paradis
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  4. Arnold L. Demain
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  5. James C. Weaver
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Gift, E., Park, H., Paradis, G. et al. FACS-based isolation of slowly growing cells: Double encapsulation of yeast in gel microdrops. Nat Biotechnol 14, 884–887 (1996). https://doi.org/10.1038/nbt0796-884

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  • DOI: https://doi.org/10.1038/nbt0796-884

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