[vc_column width=”1/1″][vc_column_text]This application note shows microtubule depolymerisation by cold during C. elegans embryo first division thanks to CherryTemp precise temperature shifts .
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Experimental conditions
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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.
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Images representative of timelapse images of the 1st division of C. elegans embryo movie[/vc_column_text][vc_column_text]
Observation
[/vc_column_text][vc_icon_text title=”At 5°C = depolymerization of the microtubules (in green), less fluorescence observed. ” icon=”krown-icon-up-circle2″ style=”two” target=”_self”]
[/vc_icon_text][vc_icon_text title=”As soon as the temperature goes back to 22°C, the microtubule polymerize again and the fluorescence intensity increased” icon=”krown-icon-ajust” style=”two” target=”_self”]
[/vc_icon_text][vc_icon_text title=”Rapid event (we were able to observe the changes within few seconds) ” icon=”krown-icon-clock-1″ style=”two” target=”_self”]
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Courtesy of Dr. Gotta and Dr. Meraldi (Gotta and Meraldi labs)
[/vc_column_text][vc_column_text][embedyt]https://youtu.be/aEN-1z4AHMA[/embedyt]
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