The air heating unit should be left ON ALL TIME.

Close the door to microscope room 1 for at least 15 min for the environment temperature to stabilize.

Temperature in the chamber can be found at the display board in the front of the air heating unit. Do not remove the temperature probe from the chamber under any circumstance.

Turn on the CO2 and the CO2 mixer ONLY WHEN live cell imaging is required.

Refill the water bottle in the chamber with MiliQ water if the water level is low.

Open the CO2 wall outlet valve (NEVER adjust the grey flow rate control):

Red handle parallel to the tube = open, perpendicular to the tube = closed.

Turn on the CO2 mixer from the power switch in the back. Wait for a few minutes to confirm the actual percentage of CO2 is steadily rising towards 5% (in case the percentage does not rise, turn off the mixer from the power switch in the back and on again).

Turn on the power supplies to the microscope, control unit and PCs from the power points on the wall, then turn on the PCs following the labeled numerical order (i.e. main power switch => Linux PC => Windows PC

Lower the objective turret all the way down using the focusing knob

Log in to CentOS @ worx:cn63501 Start softWoRx, initialize the instrument

In case the instrument does not initialize properly, please approach microscopy core staff for help.

1. CO2 supply

Use the cover with CO2 supply for live cell imaging.

Seal the sample holder if necessary (e.g. if the CO2 is believed to be insufficient because of leakages).

2. Set-up

From the SoftWoRx menu, start the acquisition software (File>Acquire(Resolve3D)). Clink YES on the dialogbox to initialize the software.

In the Acquire panel, go to Settings

Files: Under Data Folder, enter /data1/your_folder_name

This will create a folder in the main data folder (/data1), where your images will be saved.

Imaging

Save the settings and close the window

3. Running an experiment

4. File conversion