Last updated 10/29/2020
3
F20 User Guide (starting your session)
1.
Make sure microscope is in good condition:
a.
HT and FEG are on
i.
Setup → “High Tension” and “Operate” are
yellow
b.
Column valves are closed
i.
Setup → “Col. Valves Closed” is yellow
c.
EM column vacuum is stable (IGP1 = 6)
d.
Microscope left in SA mag range
e.
Stage neutral
i.
Search → click “Holder” to zero out the X, Y, Z
and α coordinates of the stage
f.
Cold trap already filled with LN
2
(can last for 16 hours);
refill every 1 - 2 hours
2.
Login to leginon workstation with your LDAP account
3.
Transfer cryo holder from the pumping station to transfer
station; cool down holder and insert your sample
a.
Dry Pumping Station User Guide (for cryo holders)
for detailed instructions
4.
Before insertion,
a.
Cryo Holder Insertion/Retraction Guide for F20
detailed instructions
b.
CHECKLIST before insertion:
1)
turbo is on and ready (yellow)
2)
objective aperture is out
3)
flu screen of microscope is down
4)
shield of cryo holder is closed
5)
airlock pump time = 120s
6)
reset holder
7)
alpha = -60°
5.
Insert cryo holder into EM.
6.
After insertion, wait for IGP1 to reach <10 before opening the column valves
7.
Select the camera/acquisition type needed for your session today:
a.
If you plan to screen multiple cryo grids today, use the F416 camera (
i.
This camera has faster throughput and yields higher contrast images in a single
exposure
ii.
If your grid looks good, you can switch to the DE20 for data collection
b.
If you plan to collect data for 2D/3D analysis, use the DE20 camera (“Cryo Data
Collection with the DE20” guide)
i.
This camera has slower throughput, but can give you higher resolution details
within multiple frames of a movie
Содержание TF20
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