FACS-Based Isolation of Slowly Growing Cells: Double Encapsulation of Yeast in Gel Microdrops

Nature Biotechnology, July 1996, vol. 14, pp.884-887

Elizabeth A. Gift, Hyun J. Park, Glenn A. Paradis, Arnold L Demain, and James C. Weaver

Isolating hyperproducing cells is important in biotechnology, but the cells usually grow slowly end 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 hr. incubation, fluorescent activated cell sorting was used to analyze and sort dGMDs with small microcolonies (slow growers) from dGMD with large microcolonies (rapid growers). Successful isolation of slow growers from a mixed population of predominantly rapidly growing Saccharomyces cerevisiae cells was achieved.

Back to Publications Page