Fast microfluidic temperature control for high resolution live cell imaging

• Published in: Lab on a chip Vol. 11; no. 3; pp. 484 - 489
• Main Authors: Velve Casquillas, Guilhem, Fu, Chuanhai, Le Berre, Mael, Cramer, Jeremy, Meance, Sebastien, Plecis, Adrien, Baigl, Damien, Greffet, Jean-Jacques, Chen, Yong, Piel, Matthieu, Tran, Phong T
• Format: Journal Article
• Language: English
• Published: England 07.02.2011
• Subjects: Cell Cycle; Devices; Dimethylpolysiloxanes - chemistry; Genes; Green Fluorescent Proteins - analysis; Imaging; Mathematical models; Microfluidic Analytical Techniques - instrumentation; Microfluidic Analytical Techniques - methods; Microfluidics; Microtechnology - methods; Microtubules; Mutations; Nylons - chemistry; Proteins; Schizosaccharomyces - cytology; Temperature; Temperature control
• ISSN: 1473-0197; 1473-0189
• DOI: 10.1039/c0lc00222d

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.