title: Fast microfluidic temperature control for high resolution live cell imaging.
abstract: One major advantage of using genetically tractable model organisms such as the fission yeast Schizosaccharomyces pombe is the ability to construct temperature-sensitive mutations in a gene. The resulting gene product or protein behaves as wildtype at permissive temperatures. At non-permissive or restrictive temperatures the protein becomes unstable and some or all of its functions are abrogated. The protein regains its function when returning to a permissive temperature. In principle, temperature-sensitive mutation enables precise temporal control of protein activity when coupled to a fast temperature controller. Current commercial temperature control devices do not have fast switching capability over a wide range of temperatures, making repeated temperature changes impossible or impractical at the cellular timescale of seconds or minutes. Microfabrication using soft-lithography is emerging as a powerful tool for cell biological research. We present here a simple disposable polydimethylsiloxane (PDMS) based microfluidic device capable of reversibly switching between 5 °C and 45 °C in less than 10 s. This device allows high-resolution live cell imaging with an oil immersion objective lens. We demonstrate the utility of this device for studying microtubule dynamics throughout the cell cycle.
authors.affiliation: Institut Curie, UMR 144 CNRS, Paris, 75005, France.
authors.forename: Guilhem
authors.initials: G
authors.lastname: Velve Casquillas
authors.forename: Chuanhai
authors.initials: C
authors.lastname: Fu
authors.forename: Mael
authors.initials: M
authors.lastname: Le Berre
authors.forename: Jeremy
authors.initials: J
authors.lastname: Cramer
authors.forename: Sebastien
authors.initials: S
authors.lastname: Meance
authors.forename: Adrien
authors.initials: A
authors.lastname: Plecis
authors.forename: Damien
authors.initials: D
authors.lastname: Baigl
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Greffet
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Matthieu
authors.initials: M
authors.lastname: Piel
authors.forename: Phong T
authors.initials: PT
authors.lastname: Tran
mesh_terms: D002453:Cell Cycle; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000032:analysis; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D055616:Microtechnology / Q000379:methods; D008870:Microtubules; D009757:Nylons / Q000737:chemistry; D012568:Schizosaccharomyces / Q000166:cytology*; D013696:Temperature
chemical_list: D004129:Dimethylpolysiloxanes; D009757:Nylons; C085707:poly(dimethylsiloxane)-polyamide copolymer; D049452:Green Fluorescent Proteins