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Microfluidics for single-cell lineage tracking over time to characterize transmission of phenotypes in Saccharomyces cerevisiae.

STAR protocols (2020-12-31)
Poonam Bheda, Diana Aguilar-Gómez, Igor Kukhtevich, Johannes Becker, Gilles Charvin, Antonis Kirmizis, Robert Schneider
ABSTRACT

The budding yeast Saccharomyces cerevisiae is an excellent model organism to dissect the maintenance and inheritance of phenotypes due to its asymmetric division. This requires following individual cells over time as they go through divisions to define pedigrees. Here, we provide a detailed protocol for collecting and analyzing time-lapse imaging data of yeast cells. The microfluidics protocol can achieve improved time resolution for single-cell tracking to enable characterization of maintenance and inheritance of phenotypes. For complete details on the use and execution of this protocol, please refer to Bheda et al. (2020a).

MATERIALS
Product Number
Brand
Product Description

Millipore
D-(+)-Raffinose pentahydrate, ≥99.0%, suitable for microbiology
Sigma-Aldrich
D-(+)-Galactose, BioXtra, pH 5.0-7.0 (20 °C, 1 M in H2O), ≥99%
Sigma-Aldrich
D-(+)-Glucose, ≥99.5% (GC)