title: Continuous high-resolution microscopic observation of replicative aging in budding yeast.
abstract: We demonstrate the use of a simple microfluidic setup, in which single budding yeast cells can be tracked throughout their entire lifespan. The microfluidic chip exploits the size difference between mother and daughter cells using an array of micropads. Upon loading, cells are trapped underneath these micropads, because the distance between the micropad and cover glass is similar to the diameter of a yeast cell (3-4 μm). After the loading procedure, culture medium is continuously flushed through the chip, which not only creates a constant and defined environment throughout the entire experiment, but also flushes out the emerging daughter cells, which are not retained underneath the pads due to their smaller size. The setup retains mother cells so efficiently that in a single experiment up to 50 individual cells can be monitored in a fully automated manner for 5 days or, if necessary, longer. In addition, the excellent optical properties of the chip allow high-resolution imaging of cells during the entire aging process.
authors.affiliation: Molecular Systems Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen.
authors.forename: Daphne H E W
authors.initials: DH
authors.lastname: Huberts
authors.forename: Georges E
authors.initials: GE
authors.lastname: Janssens
authors.forename: Sung Sik
authors.initials: SS
authors.lastname: Lee
authors.forename: Ima Avalos
authors.initials: IA
authors.lastname: Vizcarra
authors.forename: Matthias
authors.initials: M
authors.lastname: Heinemann
mesh_terms: D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D004718:Saccharomycetales / Q000166:cytology*; D012825:Silicon / Q000737:chemistry
chemical_list: D012825:Silicon