Flow cytometric isolation of growth rate mutants: a yeast model

R. Nir, R. Lamed, E. Sahar, Y. Shabtai

Research output: Contribution to journalArticlepeer-review

Abstract

A rapid flow cytometric method for the isolation of growth rate mutants was developed. The method involves single cell entrapment within small agarose beads, microcolony formation and flow cytometric scanning and sorting of the beads based on the size of the microcolony they contain. Slow- and fast-growing yeast cells could be differentiated based on the size of the microcolonies they form. The potential of this method was first verified in a model system where a minority of cells of a fast-growing yeast strain, Candida pseudotropicalis IP-315, was enriched >250-fold from an initial mixture with a slow-growing yeast, Saccharomyces cerevisiae 352-1C. Microcolony size was reliably monitored by light scatter or light absorption signals. In addition, several slow-growing mutants of C. pseudotropicalis were isolated from a UV mutagenized population. This method could be generally applicable to other colony-forming microorganisms such as bacteria.

Original languageEnglish
Pages (from-to)247-256
Number of pages10
JournalJournal of Microbiological Methods
Volume14
Issue number4
DOIs
StatePublished - Jan 1992

Keywords

  • Candida pseudotropicalis IP-315
  • Entrapment
  • Flow cytometry
  • Growth rate mutant
  • Microcolony
  • Saccharomyces cerevisiae 352-1C
  • Yeast

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