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.

Cell cycle control of spindle pole body duplication and splitting by Sfi1 and Cdc31 in fission yeast

“…Fast temperature shift of the cdc31-8 mutant was performed under the microscope (Velve Casquillas et al, 2011). The cell cycle stage at the time of temperature shift was defined according to microtubule organization, using the fluorescent tubulin marker GFP-Atb2 (Atb2 is also known as Tub1) (Fig.…”

A drug-compatible and temperature-controlled microfluidic device for live-cell imaging

“…The temperature control system used in our devices consists of a large chamber (7 × 16 mm) through which a flow of water pre-thermalized by Peltier elements is constantly maintained [33]; this chamber is positioned above the cell compartment (figure 6 a ). It was produced by cutting 81 µm thick double-sided tape (ARcare 90445) using a 60 W CO 2 laser cutter (Speedy 100, Trotec, Austria).…”

Microfluidic reactors for visible-light photocatalytic water purification assisted with thermolysis

“…29,30 Moreover, many welldeveloped microfluidic manipulation and detection techniques could be incorporated into the microreactors for in-line monitoring of the intermediate and final products. 16,30,45 V. CONCLUSIONS A photocatalytic microreactor system was constructed by directly mounting a blue-light LED panel on a microfluidic planar reactor, which enables to make use of both the heat and the light energy for organic degradation. Experimental studies have shown that the microreactor facilitates the control of photocatalytic process by adjusting the LED light intensity and the flow rate.…”

“…[5][6][7] The great success of microfluidics in many applications such as chemical analysis, biomedical assay, and drug screening has well demonstrated its remarkable capabilities in dealing with particles, fluids, and light in an integrated platform. [7][8][9][10][11][12][13][14][15][16] Thanks to its merits of flexible flow control, large surface-area-to-volume ratio, and compact size, the microfluidics could benefit the photocatalysis [16][17][18][19][20] in various aspects such as fast mass a) E-mail: apzhang@polyu.edu.hk. Tel.…”