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Microtubule depolymerisation by cold during C. elegans embryo first division

This application note shows microtubule depolymerisation by cold during C. elegans embryo first division thanks to CherryTemp precise temperature shifts .

Experimental conditions

Strain was cultured at 22°C using standard methods (Brenner, 1974). The strain used was TH65 (YFP::α-tubulin; Schlaitz et al., 2007).

For live imaging, embryos were cut from gravid hermaphrodites in egg buffer (118 mM NaCl, 48 mM KCl, 2 mM CaCl2, 2 mM MgCl2, and 25 mM Hepes, pH 7.5) and mounted on 2% noble agar pads.

Divisions were recorded using a microscope (DM6000; Leica) equipped with epifluorescence and DIC optics and a camera (DFC 360 FX; Leica). Images were collected every 15 sec using a 63×/1.4 NA objective and LAS AF software (Leica). Embryos were imaged at 22°C and the temperature was shifted to 5°C at 0sec. After 195 sec at 5°C,  the temperature was shifted back to 22°C. Temperature control and tempertaure shifts were performed using Cherry Temp.

Images representative of timelapse images of the 1st division of C. elegans embryo movie


At 5°C = depolymerization of the microtubules (in green), less fluorescence observed.

As soon as the temperature goes back to 22°C, the microtubule polymerize again and the fluorescence intensity increased

Rapid event (we were able to observe the changes within few seconds)


Courtesy of  Dr. Gotta and Dr. Meraldi (Gotta and Meraldi labs)


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