Chapter 8.

82
FEI Nova NanoSEM 650
Scanning Electron Microscope
(fei-sem) (SDH 131)
1.0 Equipment Purpose
1.1 The FEI Nova NanoSEM 650 Scanning Electron Microscope is a high
resolution microscope capable of resolving 1.4 nm features on samples
ranging in size from small pieces to 6-inch wafers. It has a number of
advanced detectors that can image highly charging samples.

1.2 This document describes important information pertaining to the FEI-SEM as

well as basic operating procedures to image samples.
2.0 Material Controls & Compatibility
2.1 The following materials are allowed in the FEI-SEM: standard semiconductor
materials, non-magnetic metals, hard-baked photoresist. Exceptions to this
list are any delaminating/flaking/peeling or other particle producing samples.
If you have any doubts, ask a staff member for approval of your material.
2.2 Substances that out-gas, have poor vacuum compatibility, or can form loose
particles in the SEM are not allowed in the chamber. If you have any doubts,
ask a staff member for approval of your material.

2.3 Magnetic samples must have staff approval before use. These samples need
special mounting to prevent them from getting pulled up into the column.

2.4 Carbon / copper / aluminum tapes, pastes, and paints are not allowed in the
chamber. Under special circumstances you may request staff approval for
the use of non-mechanical sample mounting.
3.0 Training Procedure & Applicable Documents
3.1 Specialty Tool
3.1.1 This tool is a Radiation Producing Machine. In order to fully comply
with all campus and state regulations regarding radiation producing
machines, you are required to read the Standard Operating Procedure
(SOP) associated with this tool.
3.2 FEI Microscope Systems Safety Manual
3.3 FEI-SEM Instruction Manual located in 131 of Sutardja Dai Hall.
3.4 http://www.fei.com/resources/
3.5 An Introduction to Electron Microscopy at http://www.fei.com/introduction-to-
electron-microscopy/
 3.6 ProSEM automated measurement software is available in the gowning room
for offline analysis of your SEM images.

4.0 Definitions & Process Terminology

4.1 Backscattered Electrons (BS, BSE) – Electrons that originate from the SEM
electron beam that are reflected from the sample by elastic scattering. These
electrons have higher energies than secondary electrons and often provide
good image contrast between different elements within an imaging field.
4.2 Column – A portion of the SEM that contains the electron gun,
electromagnetic lenses, apertures, and pole piece. The column is self-
contained from the main chamber and is held under high vacuum (~3.5x10 -9
torr).
4.3 NavCam – A camera that is used to assist the operator in navigating and
placing the electron beam at a specific location of interest on a sample.
4.4 Pole Piece – The very end of the column where the electron beam is
released into the chamber (see Figure 1). Never contact the pole piece with
the operator’s hands, samples, and/or sample holders.
4.5 Secondary Electrons (SE) – Electrons that are generated from the sample
as a result of ionization from the SEM electron beam. These electrons have a
lower energy than backscattered electrons.
4.6 Stigmation - Stigmation is a procedure that adjusts the beam until it is
circular. When a beam is improperly stigmated it will be elliptical leading to
images that have sharp edges in one direction and fuzzy edges in the other.
One sure way to see if your beam needs stigmation is to go through focus on
a small feature. Stigmation needs to be adjusted if the feature stretches in one
direction when it is under focused, and then stretches in the orthogonal
direction when it is over focused.
4.7 Working Distance (WD) – The distance between the bottom of the pole piece
and the focal point of the electron beam.
4.8 xT Microscope Control – User interface software that is used to conduct
imaging operations and controls on a sample.
5.0 Safety
5.1 Tool Qualification Policy - User Qualification
5.1.1 Supervised operation of the FEI-SEM may only be performed by
individuals who have completed sections 5.1.3, 5.1.4, and 5.1.5.
5.1.2 Unsupervised operation of the FEI-SEM may only be performed by
FEI qualified individuals who have fulfilled sections 5.1.3 to 5.1.10.
 5.1.3 Nanolab members must be qualified and demonstrate proper use of
the Zeiss-SEM before beginning the training program for the FEI-
SEM.
5.1.4 Read and understand the FEI Nova NanoSEM 650 Manual (this
document).
5.1.5 Attend two, half-day class/demonstrations that are scheduled by
demand. Sign up for the class in Mercury Web under the name
“feiclass”. Upon successful completion of the training classes, the
potential user is ready for hands-on training with other qualified
users.
5.1.6 Acquire 5 hours of supervised, hands-on experience from users
already qualified on the tool.
5.1.7 Pass the FEI Nova NanoSEM 650 online test.
5.1.8 Get signed off by Joanna Bettinger (joannabettinger@berkeley.edu)
by demonstrating acceptable knowledge and use of the tool.
5.1.9 To maintain a current FEI qualification, users will need at least three,
1-hour minimum sessions on the FEI every 6 months. If not met, the
user will have to attend one of the half-day class/demonstrations.
5.1.10 Users who did not work at all on the FEI for 6 months, will have to
complete the training program again, as described above.

5.2 Tool Qualification Policy - Super-user Qualification

5.2.1 To become a super-user, lab members must complete general user

training as described in section 5.1 above and the additional
requirements below.

5.2.2 Demonstration of advanced skills and knowledge specific to the

operation of the FEI-SEM.
5.2.3 Acquire more than 30 hours of hands-on operation.
5.2.4 Weekly use of the FEI-SEM for a minimum of one month prior to
requesting super-user qualification.
5.3 Reservation Policy
5.3.1 To address high demand on the FEI-SEM, reservation rules are as
follows:
The FEI may be reserved up to seven days in advance, no more
than 8 times per week for a maximum of 8 hours total. During
business hours (8:00 am - 5:00 pm, M - F) in 2-hour increments.
After hours, two consecutive 2-hour sessions may be reserved.
5.4 Personal Safety
 5.4.1 It is your responsibility to identify and understand all safety precautions
before using the FEI-SEM.
5.4.2 Nanolab members may work in room 131 alone on the SEM.

5.4.3 Under no circumstances does a member working in room 131 count as

a buddy for members in the Marvell Nanolab. There must be two
members in the Marvell Nanolab at all times. Members working on the
FEI-SEM in room 131 cannot be included towards this count.

5.4.4 No food or drink is allowed in room 131.

5.4.5 Always wear gloves when operating the FEI-SEM.

5.4.6 Always wear booties to cover your shoes prior to entering room 131.

5.4.7 Use caution closing the chamber door. Close the door slowly and
keep body parts clear of this pinch point.

5.4.8 Beware of electrical shock hazard, especially high voltage in the SEM
column and adjoining power supply.

5.4.9 An oxygen concentration monitor is located on the wall, directly

opposite the entrance. This monitor will alarm if the oxygen
concentration in the room ever gets too low (due to displacement by a
large nitrogen leak.) If the monitor alarms (beeping and flashing red
light), evacuate the room and contact NanoLab staff.
5.5 Tool Precaution
5.5.1 Points of Cleanliness
5.5.5.1 Always don booties and nitrile gloves prior to entering the FEI-
SEM room 131.
5.5.5.2 Wear clean poly or nitrile gloves on top of nitrile gloves when
touching anything that will go into the vacuum system,
including your sample and sample holders. Grease and oils
accumulate in the column and reduce the base pressure and
image quality. If your sample is contaminated with oil, wash it
in acetone and allow sample to dry prior to loading.
5.5.5.3 Report broken wafers or lost samples to staff immediately:
wafer bits left in the chamber can fall into the turbo-pump
intake or damage the stage motors.
5.5.5.4 Keep the area around the column free of clutter, and never lay
any object against the column.
5.5.2 Venting the Chamber
 5.5.2.1 It is important to minimize the length of time the chamber is at
atmospheric pressure. Do not leave the chamber vented (or
the chamber door open) for extended periods of time. When
the chamber is not under vacuum, humidity and contaminants
enter it. Such unwanted contaminants inside the chamber
result in longer pump down times and reduce image quality.
Only vent and open the chamber when you are ready to load
samples.
5.5.3 Sample Holders
Samples should be held by mechanical means in all possible
situations.
5.5.3.1 If a sample can’t be mounted onto a holder by mechanical
means contact staff for assistance.
5.5.3.2 The high resolution, three-piece sample holder is the most
common means of loading samples. This sample holder
consists of a cross screw with five holes for mounting sample
stubs, thrust washer, a threaded two-piece adjustable cylinder
which sets the height of the cross, and a slip washer (see
Figure 5). Always use the thrust and slip washer when
tightening the cross to the stage.
5.5.3.3 If your sample is a 4” or 6” wafer, you can put it directly on a
designated wafer holder. Make sure the spring mount is
holding the wafer securely.
5.5.3.4 If you have a very small sample, you should use an individual
sample holder; it holds sample stubs that can be obtained
from the Nanolab office. Put the leg of your stub into a holder
location and gently tighten the appropriate screw slot with the
1.5 mm hex wrench (finger tight, do not over-tighten). The
individual sample holder has many places for stubs. This will
allow using the rotate axis to move between multiple samples.
If you want to look at your sample at a 90 deg. angle, use a 90
deg. sample holder.
5.5.4 Loading Samples
5.5.4.1 Slowly and gently slide open the chamber door while watching
the live chamber video feed. Make sure that as the door is
closed, no part of the sample, sample holder, or stage is
above the yellow 5mm marker on the live video feed.

5.5.4.2 Verify that the three-piece high resolution sample holder is at

the standard position as designated by the Sharpie marker
mark on the elephant (see Figure 4a and 4b).
 5.5.4.3 When mounting samples onto the sample holder, tighten the
set screw with the provided 1.5 mm hex. The set screw
should only be finger tight. Do not over-tighten the set screw.

5.5.4.4 Make sure the white Teflon isolation plate is installed so that
the stage is protected from shorting during stage bias imaging.

5.5.5 Stage Sensitivity

5.5.5.1 The stage is a mechanically sensitive component of the SEM,
please treat it with care. When mounting a holder on the
stage, never use excessive pressure or rapid motions.
5.5.5.2 When closing the SEM door, be gentle, do not slam it. Before
initializing pump down, ensure the door is completely closed
and not resting on the chamber bumper.
5.5.5.3 The stage has a wide range of motion: completely lower,
center, and zero out any stage tilt before opening and closing
the chamber door.
5.5.5.4 Never perform the Home function with sample holders or
samples loaded on the stage. Running the home function with
sample holders or samples on the stage can result in a
collision and damage to the FEI-SEM.
5.5.6 Protect the Pole Piece
5.5.6.1 Caution must be used when closing the chamber door. You
must ensure the sample will not hit the pole piece prior to
closing the chamber door completely. Use the elephant (see
Figure 1) to help you judge if you can proceed with closing the
chamber door.
5.5.6.2 Testing the sample height with the elephant can lead to
false positives! Even when the elephant indicates that the
sample is low enough to close the door, the sample can be too
high and impact the pole piece. Watching the live video feed
is the only way to determine if the sample is low enough to
completely close the chamber door.
5.5.6.3 Always monitor the live chamber video feed as you open and
close the chamber door. Make sure the camera feed does not
have a green pause icon on it and that the feed is live. As you
close the door, you will be able to view your sample entering
the chamber. Make sure the sample is low enough (below the
yellow 5 mm marker) so it does not come into contact with the
pole piece as it enters the chamber. As you view your
sample safely passing under the pole piece on the
monitor, slowly close the door completely.
 5.5.6.4 Impacting the pole piece with a sample is the number one
cause for tool down time and permanent damage that reduces
the quality of images attained on the FEI-SEM.
5.5.7 Precautions During Stage Movement
5.5.7.1 Pressing ESC will immediately stop all stage movement.
When moving your sample, always be ready to hit the
ESC key to prevent anything from contacting the pole
piece.
5.5.7.2 When making stage movements, the live video feed must be
observed and your hand must be ready to hit the ESC key to
stop stage movements. Do not allow any part of a sample,
sample holder, or stage to go above the yellow 5 mm marker.
5.5.7.3 Do not change the z-axis or height of a sample with the
computer stage controller in actual, target, or relative mode.
5.5.7.4 The z-axis should only be adjusted with the mouse controls
while observing the live video feed. When controlling the stage
with the mouse, it is useful to remember that the stage
responds to the magnitude of the mouse movement. The
stage speed is also inversely proportional to the magnification.
Since the camera view has the smallest magnification, it has
the largest stage speed maximums.
5.5.7.5 Guessing the clearance between a sample/sample
holder/stage and the pole piece is not permitted.
5.5.8 Tilted samples

5.5.8.1 When imaging samples larger than 3” in any dimension

contact PE1 for assistance.

5.5.8.2 When imaging samples below 3” in any dimension:

5.5.8.2.1 Mount the chip at the very edge of the sample

holder and load it into the stage with the chip
farthest to the right hand side. This ensures the
sample will be at the highest point when the stage
is tilted.

5.5.8.2.2 Tilt the sample first while it is at the lowest height

and then raise it in height.

5.5.8.2.3 Change the tilt in increments of 5 - 10 degrees at a

time.
 5.5.8.2.4 Keep every part of the sample and sample holder
below the 5 mm marker and always keep the
sample as the highest point closest to this mark.
5.5.9 The Link Function

5.5.9.1 The Link function adjusts the WD such that it corresponds to

the distance from the pole piece to an in-focus scanning area
on a sample. It links the focal point of the electron beam to
the surface of a sample.

5.5.9.2 The link function can be used to measure the clearance

between a sample and the pole piece. Do this by focusing on
the highest part of your sample and Linking. The generated
WD is approximately your clearance.

5.5.9.3 This method does not account for tilt and clearance between
the sample/sample holder/stage and locations higher up on
the column. Extreme care must be taken to ensure the
highest point of your sample does not contact the pole piece,
even at large working distances. At all times, you must keep
your working distance greater than 5 mm.

6.0 Process Data

6.1 A log sheet of gun pressure, chamber pressure, and emission current is kept
on file.
7.0 Available Processes, Gases, Process Notes
7.1 The FEI-SEM has a number of detectors that are used for imaging samples.
7.1.1 Everhart-Thornley Detector (ETD)

7.1.1.1 The ETD is a low-resolution detector used in Mode 1 for

sample navigation. It is located next to the pole piece and can
be viewed inside the live video feed of the chamber (see
Figure 1). CARE MUST BE USED TO ENSURE YOU DO
NOT IMPACT THIS DETECTOR WHEN LOADING OR
MOVING SAMPLES INSIDE THE CHAMBER. A potential
applied to the collection grid of the ETD attracts secondary
electrons (SE) towards it. Backscattered electrons (BSE or
BS), due to their higher energies, are more difficult to collect
with the ETD. To increase BSE collection, tilt the sample for
line-of-sight reflection of BSE towards the ETD and/or use a
less positive collection grid potential.
7.1.2 Through Lens Detector (TLD)
 7.1.2.1 The TLD is located inside the column of the FEI-SEM and is
the assigned detector used in Mode 2. Typically, the area of
interest on a sample is found using Mode 1 and then, if higher
resolution is needed, Mode 2 is engaged. The TLD in Mode
2 can collect both SE (TLDS) and BSE (TLDB). If the sample
tends to charge, or information on elemental composition is
needed, detecting BSE with the TLD is recommended. If
mode 2 does not provide enough resolution or sample
charging is preventing proper imaging of samples, contact
PE1 to assist you with more advanced detectors as
discussed in sections 8.1.3 and 8.1.4.
7.1.3 Directional Backscatter Detector (DBS)
7.1.3.1 The DBS detector may only be used by Marvell staff and
superusers specifically trained and qualified on this detector.
It is a solid state detector that can be inserted and retracted
directly underneath the pole piece. The detector has four
annular rings that can be independently controlled to collect
BSE of both high and low angle projection. This detector is
often used on highly charging samples that can not be
imaged with the ETD and TLD detectors.
7.1.4 Low Vacuum Detection (LVD)
7.1.4.1 The HELIX detector may only be used by Marvell staff and
superusers specifically trained and qualified on this detector.
The Helix detector is used to obtain high resolution images of
highly charging samples. This detector is used under low
vacuum conditions. If you have highly charging samples that
cannot be imaged with the ETD or TLD, contact PE1 for
assistance with this detector.
7.2. Plasma Cleaner
7.2.1 Under some conditions, dark square areas within the scanning area of
the sample may appear. These darkened areas are a result of carbon
contaminants depositing onto the sample and reduce the quality of
imaging. The FEI-SEM has an in-situ plasma cleaner that reduces
hydrocarbon contamination in the chamber and reduces the amount of
carbon that is deposited on samples.

7.2.2 Never plasma clean anything organic, photoresist, carbon-containing

films, spin-on glass, TEM/STEM grids, or materials that oxidize easily
like Ag.

7.2.3 Typically, sample cleaning can be achieved using a 2 minute clean.

Additional 2 minute intervals may be used if necessary. The total
cleaning time should not exceed 6 minutes.
 7.2.4 Always move the stage to Y axis position over +40mm before cleaning,
remove any stage tilt, and lower stage to the lowest Z position.

7.2.5 Always ensure that the UHR (Immersion, Mode 2) lens is switch OFF
before plasma cleaning.

7.3 Mouse Control

7.4.1 The mouse (you can also use an optional keyboard) can be used to
control most of the analog inputs such as focus, brightness, contrast,
beam shift, and magnification. The only notable exception is the HV
and spot size, which has to be entered manually. Normally three
analog parameters are prescribed to the mouse at any one time; one is
adjusted by dragging the mouse with the left, middle, and right mouse
button depressed. Moving the mouse without a button pressed does
not affect the SEM. For one-dimensional inputs like focus, only right
and left mouse motion is considered. Drag the mouse in one direction
with the button depressed to increase the value of the corresponding
parameter and drag it in the other direction to decrease it. For two-
dimensional inputs like stigmation, the x- and y-axis is controlled by the
horizontal and vertical motion of the mouse controls respectively. In
addition to this, the Manual User Interface allows the use of knobs
instead of mouse controlled operation (see Figure 3). Using these
knobs can speed up operation of the tool. Function knobs available:
Magnification and Focus, Stigmator X/Y, Shift X/Y, Brightness, and
Contrast.

7.4 Status Display

7.4.1 The bottom right corner of the xT Microscope Control window contains
information about the status of the vacuum, stage, gun, and HV. In
general, the color green indicates the system is on and functioning
correctly, red means that the system is either off or not ready, and
orange means that the system is in transition between not ready and
ready. These three indicators communicate crucial information to the
user so it is important to know exactly what they mean
7.5 Vacuum Display
7.5.1 Vacuum status is shown in the bottom right corner of the xT
Microscope Control window (see Figure 2).
7.5.2 The vacuum icon is composed of two parts; 1) the column, which is the
tall rectangle and 2) the chamber, which is below and connected to the
tall rectangle.
7.5.3 During normal operation the gun icon should always be green and
have a pressure of < 5E-9 torr. If the gun icon is not green, contact
staff.
7.5.4 During venting and pump-down the chamber icon will be orange.
 7.5.5 When the chamber pressure is below 1.5E-4 torr, the icon will turn
green. At this point the electron beam can be turned on.
7.6 NavCam
7.6.1 The NavCam is a camera-loaded arm connected to the top of the
chamber door of the FEI-SEM. When the chamber door is open and a
sample is loaded, the arm is swung out 90 degrees at which point the
stage will position itself under it. By depressing the round button on
the NavCam, a picture of your sample is taken. This picture is stored
in a quad window on the computer monitor. Once the sample is
loaded and the chamber is pumped down, clicking the mouse on a
specific part of the sample in this image will automatically align this
selected area under the column. This feature should only be used with
the sample in the lowest z-position (farthest away from the pole piece)
and zero tilt. USE CAUTION NOT TO MOVE THE SAMPLE INTO
THE POLE PIECE, DETECTOR OR CHAMBER WALLS, AS THIS
WILL DAMAGE THE FEI-SEM. BE READY TO PRESS THE ESC
KEY TO IMMEDIATELY STOP STAGE MOVEMENT.
7.7 Active Data Bar
7.7.1 The data bar is located along the bottom of your active imaging quad
and contains a number of customizable fields such as horizontal field
width (HFW), magnification, high voltage, etc.
7.7.2 Double clicking on the scale bar shows all image information.
7.7.3 Users may choose which fields are displayed within the data bar by
selecting Tools, Preferences, and then the Databar tab.
7.7.4 The data bar on a saved image from a previous imaging session can
also have its fields modified. Open the image in a quad and then
select Tools, Preferences, and then the Databar tab.
7.7.5 One of the fields within the active data bar contains an editable label
that is located in the bottom right corner, under the scale bar of most
images.  Due to a bug in the software, a change to the label within an
individual account affects all other user accounts.  Do not change/edit
this label for your specific needs; the user after you may be collecting
images (with an undesirable label on them) for conference
presentations, publications, and the like.   The label shall remain as
Marvell Nanofabrication Laboratory at all times.

8.0 Equipment Operation

8.1 System Start-Up
3.1.1 Enable the SEM on Mercury, the tool name is fei-sem.
 3.1.2 If not already open, double click on the shortcut for the xT Microscope
Server software.
3.1.3 When there are green circles in front of Console Devices, Motion
NODR1, and Imaging click on the Start UI button.
3.1.4 Close the xT Microscope Server Window and login with your username
and password.
3.1.5 When the XTm: Application Status window appears, click on Hide.
8.2 Venting and Loading the Chamber
8.2.1 Vent the sample chamber by turning the beam off and then clicking
Vent button under the Beam Control work page (see Figure 2). Venting
takes about 2-3 minutes.
8.2.2 Ensure there is a live video feed of the pole piece in a quad on the
computer monitor.
8.2.3 Make sure that there is not a green pause icon in this quad (if there is
a green pause icon, click once on the quad and then F6 to un-pause
the video).
8.2.4 Inspect the chamber video quad and ensure the stage is at the lowest
possible z-axis height. To lower the stage, place the mouse cursor
over chamber video quad, click and hold the center mouse button, and
drag in a downward direction.
8.2.5 GENTLY AND SLOWLY open the chamber door while you watch the
chamber video quad making sure nothing contacts the pole piece.
8.2.6 Samples and sample holders that go into the chamber and any surface
in the chamber must only come into contact with a new, clean pair of
gloves. Don a new pair of gloves over your old ones prior to loading
samples into the chamber.
8.2.7 Swing the NavCam 90 degrees to activate it. Wait for the stage to
move under the NavCam.
8.2.8 If using the three piece high resolution sample holder, ensure that the
height of the cross screw is at the standard position as designated by
the Sharpie marker mark on the elephant (see Figure 4a). If the
sample holder is at the standard position skip to step 8.2.9.
8.2.9 To adjust the height of the sample holder, loosen it by holding the
cylinder in place with one hand and turning the cross
COUNTERCLOCKWISE with your other hand.
8.2.10 The final height of the sample holder is determined by the cylinder.
Increase or decrease the length of the cylinder by turning it
counterclockwise or clockwise respectively.
 8.2.11 Once at the desired length, verify the plastic thrust washer is in
between the cross and cylinder and then use the torque wrench to
tighten the cross onto the cylinder (see Figure 5). NEVER TIGHTEN
THE CROSS WITH YOUR HANDS.
8.2.12 Load your sample onto the stage making sure to ONLY TIGHTEN
THE SET SCREWS FINGER TIGHT.
8.2.13 Place the elephant next to your sample and ensure that the
maximum height of your sample is lower than the bottom of the trunk
(see Figure 4b).
8.2.14 Take a picture with the NavCam by pressing the round silver button
located at the base the NavCam arm. You will notice an image of
your sample loaded on the stage in one of the quads on the
computer monitor. A green cross in this screen ensures successful
image capture.
8.2.15 Swing the NavCam back to its home position.
8.2.16 Ensure there is a live video feed of the pole piece in a quad on the
computer monitor.
8.2.17 Make sure that there is not a green pause icon in this quad (if there is
a green pause icon, click once on the quad and then F6 to un-pause
the video).
8.2.18 GENTLY close the chamber door as you view the live video feed. As
you close the door make sure the sample is low enough so that
it does not come into contact with the pole piece. The highest
point of your sample should be below the yellow 5mm marker.
8.2.19 Once the door is completely closed, select the Beam Control work
page and click on the Pump button under the Vacuum menu. Gently
press the chamber door closed during the first few seconds of
chamber pump-down.
8.2.20 The bottom right corner of the screen in xT Microscope Control has
an icon of the SEM. This is located next to the lock icon. The top
rectangle represents the column and the bottom region represents
the chamber. The chamber color will turn from orange (pumping
state) to green (high-vacuum).
8.3 Imaging a Sample
8.3.1 If a yellow icon indicating a 5 mm distance from the pole piece does
not appear in the quad with the live feed of the chamber, go to the
Window menu and select CCD 5 mm Marker.
8.3.2 A yellow mark labeling 5 mm from the pole piece should now appear in
the live chamber feed. The highest point of the sample should be
below this line. If not, reduce the height of your sample immediately.
 Reducing the height is performed by clicking the middle mouse button
and dragging downward in the live chamber feed quad.
8.3.3 Navigate to a location on the sample you wish to image. This can be
done by double clicking on the desired location in the NavCam quad.
Always be ready to press the ESC key to stop the stage when
performing this step. If you see that the sample is going to
impact the pole piece press the ESC key. Pressing the ESC key
will immediately stop the stage and prevent damage to the pole
piece.
8.3.3.1 Always make sure the sample is at the lowest possible height
before making movements with the navcam.
8.3.4 Bring your sample up to the yellow 5 mm marker by placing the mouse
cursor over chamber video quad, clicking and holding the center
mouse button, and dragging it in a upward direction. You should see a
yellow horizontal line and an arrow pointing up. Notice the sample
begins to slowly move upwards towards the pole piece. Pay special
attention to the highest point of your sample. If it gets to the 5mm
marker stop raising the sample by letting go of the middle mouse
button. If at any time you wish to immediately stop the stage
press the ESC button on the keyboard.
8.3.5 Once the chamber icon is green you can turn the beam on by pressing
the Beam On button under the Column menu on the Beam Control
work page. This button will turn yellow..
8.3.6 Adjust the High Voltage and Spot values under the Column menu of
the Beam Control work page. A good starting point is Spot = 2, High
Voltage = 3kV.
8.3.7 The bottom two quads are designated for the NavCam still image and
the live video feed of the chamber. The top two quads may be used
for imaging your sample. Click on one of the top quads and un-pause
it by pressing the F6 button.
8.3.8 Adjust the scan speed to HDTV by clicking on the scan preset number
2 button located in the top-right portion of the computer screen (see
Figure 2).
8.3.9 Reduce the magnification in the imaging quad to about 35X.
8.3.10 Adjust the brightness, contrast, and focus until an image appears. It
may be helpful to do this at the edge of a sample.
8.3.11 When the sample is in focus navigate to an area of interest on the
sample, increase the magnification to ~2000x, refocus, press the Link
Sample Z to Working Distance button, and switch to Mode 2 (see
Figure 2).
8.3.12 Adjust the brightness, contrast, and focus until an image appears.
 8.3.13 Increase to the desired magnification and refocus, then adjust
stigmation in the x and then y direction.
8.3.14 Click on the Lens Alignment button.
8.3.15 You will likely see your sample wobbling in your image quad. Now
press the Direct Adjustments button.
8.3.16 Click on the Stigmator Centering tab.
8.3.17 Click on the Modulator X button. You will likely see your image
wobbling.
8.3.18 Click and drag the x- and y-axis in the Stigmator Center X alignment
box to eliminate the wobble.
8.3.19 Turn off the X wobble by pressing the Modulator X button and repeat
the same procedure for the Modulator Y button and alignment box.
Close the Direct Adjustments box when you have eliminated the
wobble.
8.3.20 Finish with final focus, stigmator, contrast, and brightness
adjustments.
8.4 Scanning and Saving an Image
8.4.1 Scan your final image by pressing the 6 preset scan button (see
Figure 2). You can see the parameters set for this scan by pressing
the arrow button to the right of this. A good starting point for this is
Resolution=1024 X 884, Dwell Time = 500 ns, Bit Depth = 16 bit,
Scan Interlace = 1, Line Integration = 100, Average =1. Changes
made to the scan preset will not go into effect until the Apply button is
clicked.
8.4.2 Press F6 and wait for the scan to complete.
8.4.3 Save the scanned image by selecting the Save As option under the
File menu.
8.4.4 Save files in the Desktop\shareddata\<your folder name here>.
8.4.5 For best compatibility across all software packages, save as an 8bit
TIF or .jpg file. Files saved as 16 bit may not open in some software
packages.
8.4.6 A shortcut for the shareddata folder is on the desktop of the support
computer. Files from this directory can be copied onto a USB drive.
Perform file transfers on the support computer, this is the computer on
the right hand side of the desk.
8.4.7 USB drives should be connected to the Sheeva USB drive only.
This drive is located on the desk next to the computer monitors. Do
not connect USB drives to any other ports of the computers.
8.4.8 To eject the USB drive, open the Sheeva Manager shortcut and select
option 3 Safely Eject.
 8.5 Venting Chamber and Unloading Samples
8.5.1 When finished taking images, lower the sample to the lowest height
and switch to Mode 1.
8.5.2 Turn the beam off by pressing the Beam On button (now yellow) under
the Column menu on the Beam Control work page. The button should
turn from yellow to grey.
8.5.3 Vent the chamber by selecting the Beam Control work page and click
on the Vent button under the Vacuum menu.
8.5.4 Once vented, slowly open the chamber while monitoring the live video
feed of the chamber. Ensure that the pole piece is not brought into
contact with the sample, sample holder, or stage.
8.5.5 Remove the sample.
8.5.6 When the sample has been removed, close the chamber door
completely and pump the chamber back down by selecting the Beam
Control work page and clicking on the Pump button under the Vacuum
menu. Gently press the chamber door closed during the first few
seconds of chamber pump-down.
8.5.7 Log out of the xT Microscope Control software under the File menu.
9.0 Troubleshooting Guidelines
9.1 What is stigmation?
9.1.1 Stigmation is a procedure that asymmetrically stretches and squishes
the beam until it is circular. When a beam is improperly stigmated the
beam often will be elliptical leading to images that have sharp edges in
one direction and fuzzy edges in the other. One sure way to see if your
beam needs stigmation is to go through focus on a small feature. If the
feature stretches in one direction when it is under focused, and then
stretches in the orthogonal direction when it is over focused, stigmation
needs to be adjusted.
9.2 I am having trouble stigmating. What is the secret to getting optimum
stigmation?
9.2.1 Unfortunately stigmation is both crucial and difficult. The standard
stigmation procedure is to stigmate in the x first. Obtain the narrowest
beam in the corresponding direction, then stigmate in the y and do the
same thing. The problem is that the corresponding direction is as a
rule, not the direction you are stigmating. This can be fixed by rotating
the image so that the stigmation directions correspond to x and y. Take
your time when trying to find the optimum stigmation values, it takes
practice. Poor stigmation is the number one cause of bad images.
9.3 My image is degrading with time, what can I do?
 9.3.1 There can be many causes of this problem. For high-resolution
pictures you might need to stigmate periodically. Your sample could be
charging; gold coating the sample helps. Your sample could be getting
damaged; if you are looking at sensitive materials, try lowering the HV.
9.4 I cannot see any of my features!
9.4.1 This is a common problem for samples with small or faint features.
Remember that some things that are readily visible optically are
practically impossible to see under an electron beam. Scratching your
sample near the region of interest will help to provide identifying marks.
In general, try to find a large feature on low magnification, do a coarse
focus and only then go to high magnification.
9.5 How do I get more information on the FEI?
9.5.1 There is a lot of information under the help menu. You can also email a
super-user with your questions.
10.0 Study Guide and Super-User Check-List
10.1 Online Test
10.1.1 Users should read and understand this manual to prepare for the
online exam. Special attention should be placed on bold text and
the safety section.
10.2 Super-User Checklist
10.2.1 Super-users are to assist in the training of new users on the FEI-
SEM. Users interested in the FEI-SEM should read the manual
and gain hands-on training with super-users. The super-user
should ensure the new user has working knowledge and
experience with the following key points:
10.2.1.1 The use of blue booties and gloves prior to entering the
FEI-SEM room.
10.2.1.2 Opening and closing the chamber door slowly and gently.
10.2.1.3 Inspection of the live video feed while opening and
closing the chamber door to ensure the pole piece is
protected.
Do not leave the chamber vented (or the chamber
door open) for extended periods of time.
10.2.1.4 Materials allowed in the FEI-SEM.
10.2.1.5 Materials not allowed in the FEI-SEM.
10.2.1.6 The use of mechanical clamping for sample mounting.
10.2.1.7 Carbon / copper / aluminum tapes, pastes, and paints are
not allowed in the chamber without staff approval.
10.2.1.8 Tighten all set-screws finger tight.
10.2.1.9 Never perform the Home function with sample holders or
samples loaded on the stage.
10.2.1.10 Use of the elephant to measure the height of the three-
piece high-resolution sample mount and also the overall
sample height.
10.2.1.11 Changing the z-axis with the computer stage controller,
either in absolute, target, or relative mode, is not
allowed.
10.2.1.12 Pressing ESC will immediately stop all stage
movement. When moving the stage/sample, always be
ready to hit the ESC key to prevent it from contacting
the pole piece.
10.2.1.13 How to tell if the chamber is under vacuum or vented.
10.2.1.14 How to tell if the chamber video feed is paused or live.
10.2.1.15 Proper use of the NavCam.
10.2.1.16 How to get a yellow 5 mm marker on the live video feed
and never to use working distances less than 5 mm.
10.2.1.17 Understanding the Link function and how to use it.
10.2.1.18 Understanding the difference between imaging in Mode
1 and 2.
10.2.1.19 What the acronyms EHD and TLD mean.
10.2.1.20 Saving data in Desktop\shareddata\<your mercury login
name here>.
10.2.1.21 USB drives should be connected to the Sheeva USB
drive only. This drive is located on the desk next to the
monitors. Do not connect USB drives to any other ports
on the computer.
10.2.1.22 Use of the Sheeva Manager to safely eject USB drives.
11.0 Appendices/Figures & Schematics

Figure 1: Images of the Chamber

Figure 2: The xT Microscope Control Window

 Scanning
Lens Presets
Alignment Mode 1/2

Direct Working
Adjustment Pages

Quad 2 Quad 1

Live Chamber
NavCam Video Quad
Quad

Figure 3: The Manual User Interface

Figure 4: Elephant Height Check

Figure 4A Figure 4B
 Figure 5A

Torque Wrench

Cross Screw

Thrust
Washer

Cylinder
Slip
Washer Figure 5B

Slip
Washer

The small slip washer fits inside the hole of the larger Teflon plate.
12.0 Appendices
APPENDIX A
FEI Detector Mode Schematics
 APPENDIX B
Useful Keyboard Shortcuts
ESC – Immediately stops all stage motors
+/- – Increase / decrease magnification
F2 – Instantly takes a photo or snapshot based on parameters set in Preferences
F3 – Pop-up linear signal for contrast and brightness adjustments
F4 – Instantly takes a photo or snapshot based on parameters set in Preferences
F5 – Makes selected quad window full screen view
F6 – Stop / start, pause / unpause
F7 – Jumps to small screen view for higher refresh rates during focus and stig
adjustments
F12 – Enables stage rotation

Tab+<1-6> - Switches between scan mode presents 1- 6

Crtl+R – Resets scan
Crtl+0 – Returns stage x and y positions to 0,0 (centered under pole piece)
* – Rounds magnification to a more friendly number

APPENDIX C
Useful Preference Settings
Lower stage when venting – Full Down
Blank beam during long moves – No
APPENDIX D
Comparison of Imaging Modes

APPENDIX E
 Pump Oil Compatibility
The following is adapted from the SPI (electron microscope supplies) web site
(http://www.2spi.com/catalog/vac/santovac-5.shtml)

Santovac (pump oil) fluids are known for their low backstreaming characteristics, which
is especially important for electron microscope applications. This is the reason why it is
so highly used not only in analytical laboratory instrumentation but also in production
electronics applications. However, since the product is a hydrocarbon fluid, the
molecular species from the fluid, in the presence of the ionizing radiation from the
electron beam, will cause polymerization, contributing to the overall contamination of the
column.

For many users of electron microscopes, another alternative would be to use one of the
perfluorocarbon based diffusion pump fluids, such as Fomblin® or Krytox®. While all
diffusion pump fluids do have some low level of molecular species in the microscope
column, the perfluorinated polyether species do not get polymerized in the presence of
the ionizing radiation as is the case for hydrocarbon pump fluids. The end result of this
advantage is that the column runs cleaner, longer and whatever contamination in the
column that might otherwise be present is present but at much lower levels. Putting it
another way, microscope downtime is greatly reduced since the column runs much
cleaner for much longer periods of time.

Upon rare occasion when mechanical mounting of samples is not sufficient, pump oil
may be used (only with prior permission from staff). Fomblin, Krytox or other
perfluorinated ethers are acceptable for sample mounting, however no hydrocarbon
based oils are allowed.

J. Clarkson – April 2013

J. Bettinger - Sept. 2016