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C. elegans embryo first division temperature control

Controlling C. elegans embryo first division using the CherryTemp system

 We thank Dr. Gotta and Dr. Meraldi’s labs for sharing these results.
Visit the Gotta lab and the Meraldi lab in Gebeva!

Methods

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 thanks to the special control stage equipment. After 195 sec at 5°C,  the temperature was shifted back to 22°C. Temperature control and tempertaure shifts were performed using CherryTemp.

Results

Representative timelapse images of the 1st division of C. elegans embryo.

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).

CherryTemp heater/cooler for live-cell imaging

Dynamic & fast : 10 seconds temperature shifts


Stable & Precise : Long term stability


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