Isolation of single yeast cells by optical trapping

Jos A. Grimbergen, Koen Visscher, Daniel S.Gomes De Mesquita, G. J. Brakenhoff

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

Individual yeast cells can be successfully isolated and recultured on plates with a new isolation method making use of optical trapping with infrared laser light. The cells can be selected on morphological criteria by high resolution microscopy. The isolation device is constructed from two coverslips separated by spacers, in which selected cells are transferred to a plastic capillary, using the optical trap. To test the procedure, selection experiments were done with a mixture of two Saccharomyces cerevisiae strains, distinguishable both in fluorescence microscopy and on agar plates. These experiments showed that only selected cells were isolated, and close to 100% of the isolated stationary‐phase cells formed colonies on agar plates, indicating a high recovery. A lower recovery was obtained with exponential‐phase cells, possibly because of a higher sensitivity to laser irradiation. Applications for this method may include the isolation of mutants with altered morphology and the isolation of subpopulations of yeast cultures, for their separate investigation or for the initiation of pure cultures.

Original languageEnglish (US)
Pages (from-to)723-732
Number of pages10
JournalYeast
Volume9
Issue number7
DOIs
StatePublished - Jul 1993
Externally publishedYes

Keywords

  • Optical trapping
  • isolation method
  • yeast

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Genetics

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