Professional Documents
Culture Documents
Training SOP
NanoTech User Facility (NTUF)
Center for Nanotechnology
University of Washington
June 2014
Gun = 1
Column = 6
Camera = 31-ish
d. Check that the stage has been re-centered – double check both the numbers
Stage tab
at the bottom of the software interface and in the Stage tab.
e. Cover the glass viewing chamber (both covers), then fill the nitrogen trap
(anti contamination device or ACD). PPE is provided. Liquid nitrogen transfer
dewars can be filled with the 50 L tank in the hallway near G44M entrance.
Lens tissue
Holder Tool
Double Tilt Sample Holder – you must be specifically trained
in order to use this holder!
Important: beryllium is VERY toxic and special care must
always be taken when working with this holder!
Do not push down or apply force to the carrier at all or the pivots will break. The pivots are mounted on fragile crystals that
calibrate the -tilt.
a.Use the Hex Tool to unscrew the retaining nut and lift it gently out of the carrier (DO NOT use tweezers – use the hex tool).
b.Place grid in carrier and check alignment on the stereo microscope.
c.Put the washer on top of the grid with the tabs aligned if a thin grid or sample is being loaded. If you have a thicker sample, do
not use the washer. Always store the washer when not being used in the small plastic box labeled for it.
d.Lift the retaining nut on the Hex Tool and carefully screw it into the carrier (again, DO NOT use tweezers – use the hex tool).
e.Test that the grid is clamped firm and not loose in the carrier.
*attention…always use the vacuum tweezers when handling beryllium parts…this alleviates the
possibility of contaminating tweezers with beryllium. If you touch beryllium with a gloved hand
accidently, remove and throw away the glove without touching it on anything else.
Helpful Tips:
Hex Tool •Keep small parts close to the table top in case they fall.
•Don’t over tighten the nut. You should get about 1 full turn
on the threads and the nut will be recessed into the carrier.
Washer
•Take care not to insert the retaining nut upside down.
There is a flange around the top of the nut.
Insert sample holder
a. Before inserting go to “Stage” tab and ensure the stage has been reset.
b. Open “Setup” Tab and “Vacuum Overview” in the lower right hand bar of software.
c. Align pin to 4 o’clock position, and insert TEM holder straight into the airlock with a
d. slight clockwise turn until it sets in.
e. “Turbo On” button will turn from gray to Orange and a message will pop up below
Setup tab
f. for holder selection. Select holder type in message box and click Enter (arrow) button.
g. “Turbo On” button will change from Orange to Yellow and once it is yellow will go
h. through a series of pumping cycles for three minutes. Red LED on the
i. goniometer (stage) will light up.
j. Watch vacuum status change to “Airlock.” V41 & V8 will show as open on Vacuum Overview.
k. Once Red LED on the goniometer (stage) turns off and vacuum status changes back to
l. “Col Valves Closed”, the holder can be inserted further into the goniometer:
m. - Turn the holder counter-clockwise to the stop, hold it (without pulling the holder or resisting),
Vacuum
n. and gently guide it in. DO NOT let go since it may slam against the goniometer and can cause status
window
o. damage to the electronics behind the purple plate.
Setup tab
Left-hand control panel (LH) Beam will be visible in chamber Right-hand control panel (RH)
TEM Alignment: Condenser
4 cm ring on phosphor
X-Align
Y-Align
Selector
dial
C2 Aperture
TEM Alignment: Condenser Stigmation
a. IF the area of illumination is not circular or the beam
appears to stretch at the intensity cross-over then perform
the following:
b. Go to “Tune” tab > “Stigmator” menu and click on
“Condenser.”
c. Use the “Multifunction Knobs” (MF) to adjust the beam
roundness in both the X and Y dimensions. Check that the
beam does not stretch at the “Intensity” cross-over.
d. Click “None” after finishing.
Tune tab
Multi-Function X Multi-Function Y
Main phosphor
screen up/down
TEM Alignment 5: Objective Stigmation and Focus
a. Go to “Process” tab in Digital Micrograph program and select
Live > FFT.
b. IF the FFT image is not circular go to the “Tune” tab >
“Stigmator” menu and click “Objective.”
c. Turn MF knobs to correct any objective astigmatism and make
the FFT image circular.
d. Click “None” after finishing.
e. Adjust “Focus” to maximize the diameter of the rings which
puts you near minimum contrast focus.
f. Defocus image counter-clockwise to make edges appear
sharper.
g. Click “Acquire” to capture the focused micrograph.
h. Optional: at minimum contrast focus push “L1” on LH control
panel to reset defocus reference point.
Tune tab
NOTE: the cryo cycle can NOT be disrupted during the cycle…it must run the entire time.