Video - Methodology description linked to Jacob Hanna lab manuscript - Aguilera-Castrejon et al. Nature 2021:

https://rdcu.be/cgWj2​

https://dx.doi.org/10.1038/s41586-021...​

Part 1. Embryo dissection. Step by step visualization of material and protocol for dissecting E7.5 embryos for ex utero culture. These guidelines apply also for E5.5 and E6.5 embryos.

Part 2. Media preparation and pre-heating. Detailed visualization of protocol for preparing ex utero culture media and pre-heating inside the roller culture.

Part 3. Opening the gas regulation module and operating the incubator’s gas flow valve. Part 1. Opening of the gas regulation module to show the internal components and pressure transmitter which controls the gas pressure depending on the Voltage. Part 2. Setting the proper gas flow by turning the gas flow valve and monitoring the rate of bubbles created in the gas outlet tube. Turning the valve counterclockwise results in the absence of gas flow and clockwise turning increases the bubble flow rate.

Part 4. Embryo transfer to a new culture bottle. Transferring of one embryo to a bottle containing freshly prepared pre-heated media.

Part 5. Embryo imaging. Transferring of 3 embryos to a plate for imaging under a stereomicroscope.

Movie 6. Embryo culture in static plates and transfer to the roller culture. Embryos explanted at E5.5/E6.5 are cultured in a CO2 incubator in individual wells of an iBidi plate and subsequently transferred to a bottle for roller culture.