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Isolation of single yeast cells by optical trapping
Author(s) -
Grimbergen Jos A.,
Visscher Koen,
De Mesquita Daniel S. Gomes,
Brakenhoff G. J.
Publication year - 1993
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320090706
Subject(s) - biology , yeast , isolation (microbiology) , agar , agar plate , microscopy , fluorescence microscope , saccharomyces cerevisiae , trapping , biophysics , microbiology and biotechnology , fluorescence , optics , biochemistry , bacteria , genetics , ecology , physics
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.