SEM Standard operating procedure

Doug Kim/ Chun-Min Feng, 09/19/2005

Sample Preparation
1.

Place your samples on the aluminum holder stub using a double sticky carbon
tape
Insulating samples have to be located with either carbon or gold and electrically
grounded. Gold coating is much conductive, but not good for EDX analysis.
Also, use silver paint to electrically ground the sample. Conductive samples do
not need this process.
If you have many samples look alike, inscribe marks on the sample stub or
samples. It is very hard to recognize similar samples in the SEM.
Then, completely dry the sample in the drying oven at 60C for at least 3 hours
depending on the sample conditions. The rule of thumb is that it is better leave
them overnight in the drying oven.

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Loading the sample into the SEM holder

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Make sure that the valves of the two nitrogen gas tanks are open. If theres not
enough nitrogen (less then 100 psi), please notify Jorge Morales (Electron
Microscope Centre Manager)
Press the Vent button located at the display panel of the Microscope table
After a click sound (it may take about 1 minute to fill up the chamber with
nitrogen), locate the lever on the bottom of the door and gently pull the lever up
and open the chamber door.
Move the sample mounting stage all the way down by pressing z-axis down arrow
key (located on the keyboard of the microscope control table)
Place the sample holder stubs into the mounting holes. Tall samples should be
located away from the left side of the chamber, where the secondary electron
detector exists. Using the yellow long screwdriver tighten the set screws for the
mounting holes. Double check whether the sample holders are well tightened.
Make sure nothing gets in the way to close the door. Gently, close the door and
then press the EVAC button.
Now you will hear the sound of rotary pump. Wait for about 2 minutes to see you
get the green light on the display.
Wait about 30-45 minutes to achieve high vacuum < 5 X 10-5 Pa

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Turning on the SEM

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When vacuum reaches proper level, filament light will be ON.

Turn the key switch to ON position. Now the monitor will be ON.

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Check the status of dial positions. The acceleration voltage should be 15 KV

and spot size should be Large and 1.
Turn on the filament and check if you get the red light. If you did, the filament
is burned out. (Contact Jorge Morales). Now turn on the high voltage.
Advanced users: check the filament emission, beam alignment, aperture
alignment, and astigmatism.
Go to the lowest magnification (10X). Choose TV scan mode.
Find your samples using trackball
Turn on the coarse focus switch. Using the focus knob, change the working
distance to 14 mm. Do not get closer than 6 mm. Your sample may bump into
the objective lens.
Now bring up the sample stage slowly by pressing z-axis UP key. In the
meanwhile, look the screen and find the z-position where the image is in
focus. Write down the z-position in your lab notebook.
Turn off the coarse focus [Coarse: light off; Medium: light on]
Switch to the slow scan mode and increase magnification. If the magnification
is higher than 1000X, switch the spot size to medium. Use outer focus
(medium) ring to focus the image.
Advanced users: If necessary, increase or decrease acceleration voltage. For
EDX of some materials, acceleration voltage of 25 KV may be necessary to
generate higher energy spectrum of x-ray emission. Confer the energy
diagram of various elements pasted at the laboratory wall.
Press VARIABLE button to open up a small variable window on the screen.
Adjust the size of the screen and overlay it in the region of interest. Focus the
image within the small screen using outer focus ring and later inner focus ring
for fine focus.
Advanced users: It is strongly recommended to turn off the automatic control
of brightness and contrast. Adjust them manually by turning both dials counter
clockwise all the way down so that indicator lights are all located on left end
(red color). Start turning brightness dial clockwise until brightness indicator
start to move right. Then slowly turn contrast dial for its indicator light to
reach middle.
Make sure that the scanning speed is set to S1. Only S1 scanning speed will
allow electronic imaging acquisition.
Double click the Spirit icon on the computer.
Go to the File menu and select preferences. Select your directory under the
folder name Glen Kowach and input the sample name and photo ID number.
Click image set up icon. Select the Mapping option. Normal resolution is 1024
and frame is 1. Preview the image and adjust the contrast and/or brightness.
Click OK to close the image setup window.
Click image acquire icon to record the image. Now the software takes the
control over SEM and the monitor will freeze. Do not change any parameters
during the image recording.
Save the image as .tif format. Add the magnification in the file name.
For scale bars, go to Imaging menu and select add scale bar. Choose the length
of scale bar you want and place it where you want it to be in the image. Before

saving the image, go to the annotation and select burn in all the annotation to
finalize the scale bar into the image. Save the image file as .jpg format.
In general, smaller spot size will give you lower contrast, grainy image, smaller depth of
focus, but better resolution in the acquired image.
For the magnification of 1000X and UP using medium spot size is recommended. Small
spot size can be used on the much higher magnification (10,000 X and up).
Put line to check the signal profile. If the profile shows sharp peaks and valleys, it means
that your focus is good. If those peaks and valleys are broad, then focus is not good.
For Energy Dispersive X-ray Spectroscopic Analysis
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Set the acceleration voltage to 20 KV. The working distance should be 14 mm

Move down the detector to the 45 mm by rotating the knob below the detector.
Focus on your sample
Click X-ray setup icon. Make sure that the box of Enable is checked. Live time is
usually set for 200 secs.
Click X-ray icon. Place the cursor on the spectrum window and read the counts
per second and dead time (DT) on the bottom of spectrum window.
Adjust DT to fall in between 25 to 30 % by changing the spot size. A larger spot
size gets higher DT and counts per second. Refocus afterwards.
Click to erase current spectrum and start over collecting the X-ray spectrum.
To identify peaks, click on the periodic table. Select possibly existing elements in
your sample and see if those peaks are matching.
Save the spectrum as .pgt file.

Turning OFF the SEM

1. Lower the magnification to 10X. Set the spot size to larger and 1. Go to TV scan
mode. Set the acceleration voltage to 15 KV.
2. Bring down the sample stage all the way down so that the z-position reads 0.
3. Turn the key switch to VAC position.
4. Make sure you have enough nitrogen gas and vent the chamber to take out your
samples.
5. Close the chamber door and evacuate it.