View: bin 1, 0.2 s exposure
Record: bin 1, 5s exposure
Here’s how I set up SerialEM data collection on the Krios
Save navi
Align to marker
Go to 205x mag
Close out of montage
New montage – MMM
Add squares
Then check the MMM acquire script
Acquire at items – MMM
Find constrast-y contamination, add point, go to low dose mode, go to XYZ & shift to marker
Find square without features, set focus point. Change view to square, bin 8, 0.5s exposure & shrink. Once perfectly centred, save to buffer P.
Put objective aperture in, check stigmation.
Astigmatism by CTF, coma-free by CTF, astigmatism by CTF again.
View needs to be same as buffer P i.e. small & square
Set multi-shot parameters.
Acquire at items!
Use winSCP to copy gain ref to own workspace (although I think Sasha does this already now).
e2proc2d.py gainref.dm3 newname.mrc —meanshrink 2
Copy the new gain ref back to capture data
Using puTTy log onto taylorimages (cryoemiscool)
screen -D -RR
sudo semi.pl
Add notes if necessary
Run!
Old exposure settings – 5.9 s, 0.074 sec/frame. 80 frames total.
New exposure settings – 5s, 0.05s/frame (100 total). Total dose 70e- (0.7 e-/frame)
Taylor Lab 