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Rapid determination of plasmid‐carrying yeast cells by using an imaging sensor system
Author(s) -
Endo Hideaki,
Suzuki Masayasu,
Sode Koji,
Tamiya Eiichi,
Karube Isao
Publication year - 1991
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260381111
Subject(s) - plasmid , yeast , fluorescence microscope , recombinant dna , biology , fluorescence , microbiology and biotechnology , chromatography , chemistry , biochemistry , gene , optics , physics
A novel imaging sensor system for the determination of plasmid carrying yeast cells was developed. The sensor system consisted of an Silicon Intensifier Target (SIT) video camera, a fluorescent microscope, and a personal computer system equipped with an image memory board. This system was based on the fact that the membrane integrity of only plasmid‐carrying cells is lost following cell growth in 5‐fluoro‐orotic acid (5‐FOA) containing medium, and consequently these target cell can be stained with fluorescent probes and detected. In this study, plasmid‐carrying cells were detected and their fraction determined in a mixture of both plasmid‐carring and plasmid‐free cells. A good correlation was observed between the values determined by this sensor system and the conventional method in the 30%‐80% range, and one assay was possible within 4 h. This sensor system could be used for the monitoring of plasmid‐carrying fraction in recombinant yeast cells during cultivation.