title: FACS-based isolation of slowly growing cells: double encapsulation of yeast in gel microdrops.
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.
authors.affiliation: Harvard-M.I.T. Division of Health Sciences and Technology, Cambridge, 02139, USA.
authors.forename: E A
authors.initials: EA
authors.lastname: Gift
authors.forename: H J
authors.initials: HJ
authors.lastname: Park
authors.forename: G A
authors.initials: GA
authors.lastname: Paradis
authors.forename: A L
authors.initials: AL
authors.lastname: Demain
authors.forename: J C
authors.initials: JC
authors.lastname: Weaver
mesh_terms: D002455:Cell Division; D002469:Cell Separation; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D012441:Saccharomyces cerevisiae / Q000166:cytology*
chemical_list: D005456:Fluorescent Dyes