Updated 5/23/19
Leica EM-GP & GP2 User Guide
● Users must wear protective gloves and goggles when handling liquid nitrogen and ethane
● Setup of the EM-GP takes 30 - 60 minutes. Please plan in advance.
● Shutdown of the EM-GP takes 1 - 2 hours. Please check if you are the last user of the day. If you are not,
please coordinate with the next user.
1. Turn the machine on (power switch at back side of machine).
2. Place the black ethane cup and the silver transfer container into the dewar chamber (usually stored inside
of fume hood).
3. Fill the humidifier with 60 mL distilled water.
4. Fill dewar chamber with LN
2
until 100% full.
a. refill every ~20 minutes
5. Once the cryogen container temperature < -175
℃
, liquefy ethane gas until black container full
a. refill 2 - 3 times as ethane will warm up container temp no longer liquefy
6. Fill silver transfer container with liquid nitrogen.
a. refill every ~5 minutes
7. Gently transfer cryo gridbox into silverver transfer container. Refill liquid nitrogen.
8. Setup environmental chamber settings
a. T
C
= 4
℃
- 25
℃
(will only go as low as 7
℃
)
b. H
R
= 90 - 99% (~15 minutes); NOTE-
humidifier will only run when T
C
is reached
c. Setup temperature of cryogen = -175
℃
9. Setup blotting parameters
a. On main screen, click the long rectangular
grey box in the upper left side (P1
___/1.0/___).
b.
Select option Nr. 1 (see image →). This is
the standard recipe used at the NYSBC. You
are welcome to make your own recipe,
but
DO NOT change Nr. 1 recipe.
c.
Select “OK” to exit screen
10.
Setup tweezer/blotter position parameters. The 2 main settings that need to be adjusted are “blotter
settings” and “grid blot position”.
a.
On main screen, select “Setup” button
b. Blotter settings
c.
Click “Adjust”
d. Lower chamber? OK
e.
Select “Blot Position”
i.
Using binoculars and adjust buttons, move blotter closer or further from grid to optimize
contact (
218
works well for us)
ii.
Select “Back” when done
f.
Grid Blot Position
i.
Click “Blot Position”
ii.
Using microscope and adjust buttons, move tweezer up or down from blotter to optimize
contact (
3.6mm
works well for us)
iii.
Select “Back” when done
g.
Select “Prepare” to exit screen
11. Freeze grid (practice first with a dummy grid and water)
