Setup for live cell
1. Turn on the main CO
The carbon dioxide is supplied from the tank opposite the Nikon Ti room. When CO
_2 is not being used the tanks should all be off and the main gauge reading 0 kPa.
2. Turn the CO
_2 regulator in the room
_2 is supplied to the incubator from the tap in the wall behind the Nikon Ti.
- Open blue tap fully
- Open the regulator. The gauge should read 20-40.
3. Check the water level in the humidifier bottle
There is a bottle inside the microscope cage on the right hand side below the stage that humidifies the air coming from incubator. Check that this half full and top up if necessary.
3. Turn on the Incubator
- Turn on the incubator on from the blue switch
- Make sure the flow rate dial is on. You should see bubbling in the humidifier bottle.
- Set the target temperature and CO
_2concentration by pressing the green/red buttons to increase/decrease the targets.
5. Attach the well plate cover
The well plate cover should be in the microscope cage attached to the humidifier bottle.
5. Wait for equilibrium
It will take some time for the conditions in sample to reach their target values.
_2- 10 min: The air above the sample will equilibrate very quickly, less than 5min.
- Temperature - 1 hour: The temperature of the sample will take longer as large volume of air in the cage needs come to equilibrium. For a 15°C difference from room temperature this takes about an hour. If you need higher temperature you will need to wait longer - check current temperature in the cage on the incubator screen.
- Stage movements - 2 hours: The microscope stage expands it warms up which shifts the sample slightly. Changes in Z can be corrected for using the PFS system, but the shift in XY position may affect your experiment. For high-resolution time-lapse imaging allow 2 hours for equilibration.
Shut down for live cell
- Turn off the incubator.
- Place well plate cover next to the humidifier bottle.
- Top up the humidifier bottle.
- Turn the regulator and tap off at the wall.
- Turn the tap on the CO