Fast microfluidic temperature control for high resolution live cell imaging

Lab Chip. 2011 Feb 7;11(3):484-9. doi: 10.1039/c0lc00222d. Epub 2010 Nov 19.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle
  • Dimethylpolysiloxanes / chemistry
  • Green Fluorescent Proteins / analysis
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods*
  • Microtechnology / methods
  • Microtubules
  • Nylons / chemistry
  • Schizosaccharomyces / cytology*
  • Temperature

Substances

  • Dimethylpolysiloxanes
  • Nylons
  • poly(dimethylsiloxane)-polyamide copolymer
  • Green Fluorescent Proteins