Yeast cold-shock microtubules depolymerization with CherryTemp

We depolymerized S. pombe microtubules while imaging the cells (gamma-tubulin GFP) with a cold shock at 5°C (25°C to 5°C in less than 10 sec). We then repolymerized by shifting back the temperature within seconds to 25°C.

Temperature shift to image S.pombe temperature sensitive mutants

Application example: Block and release of cdc25-22 S. pombe ts mutants.

Figure 1, cdc25 ts S. pombe reversible control of mitosis arrest by CherryTemp

Figure 2, CDC25 ts S. pombe reversible control of mitosis arrest by CherryTemp, We thank Damien Coudreuse’s lab (Julien Babic, PhD st. – IGDR, Rennes) for credits.

cdc25-22 mutants were blocked at the G2/M transition for 4 hours at restrictive temperature (36.5 °C). Synchronous release into the cell cycle was achieved by a quick shift (10 seconds) into permissive temperature (25°C). Septation index shows the high synchronicity in the release.

CherryTemp™ – Heater/Cooler system for Yeast