CSB CryoEM Facility TF20 Operating Instructions SEC
05/02/16
VI.
Sample Loading (cryo)
1.
Make sure the anticontaminator dewar is filled with liquid nitrogen (LN2).
2.
The cryo holder should be pumped at least 2hrs in the Dry Pumping Station (see separate
protocol) before use. Preferably, you'll have pumped the holder the day before for at least 6hrs.
3.
Place the cryo transfer station on the table top. Insert the cryo holder into the station and
remove the clip ring and previous grid if present.
4.
Make sure you have gathered your cryo sample in transfer dewar, forceps, large forceps for
grid box transfer, and anything else needed and place it near the transfer station.
5.
Add LN2 to both the cryo transfer station and the cryoholder. These will boil at different times.
Do not overfill the holder nor the station, this will only cause the holder to become icy and the
transfer station to frost.
6.
Plug in the control unit cord and turn on the temperature control unit.
7.
Tilt the compustage to 60° by going to the Search tab, flapout window, Control tab, Alpha
toggle, type in 60° and click Set Alpha. Turn the Turbo on.
8.
Check that the airlock pump time is set to 6090s (Setup tab, flapout window, Settings tab,
Default airlock time: 60s).