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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Figure 1. Evac and Display Breakers. Vent/Evacuate button is also shown. The button
should be in (yellow indicated) for vacuum and out for vent. The instrument should
normally be left under vacuum.
Figure 2. X-Ray and Robinson detectors fully retracted. The X and Y controls of the
stage should be at 30 and 20 respectively (use the numbers on the non-movable part of
the micrometer indicator). Z should be at the exchange position (white squares
touching). Tilt should be at 0. Rotation can be at any position.
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Inserting a Specimen
• Select a sample holder and stub (see Fig. 3)
o The holder is the part that fits into the stage, the stub is the part upon
which the sample is mounted
• Mount the sample on the stub (see Fig. 4)
o Usually use carbon tape or carbon paint or silver paint to adhere the
sample to the stub
• Mount the stub on the holder (see Fig. 5)
o Screw the stub into the end of the threaded rod
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Figure 3. Sample holder and stubs. The sample stub screws into the sample
holder
Figure 4. Carbon tape used to adhere sample to stub. On the right, a sample is shown
held in place on a stub by a strip of C tape. It is acceptable to use more than one strip
of tape and to put more than one sample on the stub at a time.
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Figure 5. Specimen height gauge. Loosen the locking collar on the sample holder to
adjust the sample height. The specimen height should be set to the center mark on the
gauge. This sets the working distance to 23mm. For X-ray analysis, set the specimen
height to the second mark for a working distance of 17mm.
• Turn on the Chamberscope and confirm that there are no samples currently
in the instrument and that the detectors are fully retracted.
o Turn on the Chamberscope power button
o Turn on the monitor
o Adjust the illumination control as needed
• Confirm that the Z-stage control is set to the exchange position (EX)
o The white squares will line up when the stage is at the exchange position
• Press the EVAC/AIR button (see Fig. 1) to bring the chamber to atmospheric
pressure.
o The LOW light will turn on
o The system will reach atmospheric pressure in approximately two minutes
• Carefully open the chamber
o The stage will slide forward
• Insert the sample holder into the stage
o Observe that the sample will not strike any detectors or the pole piece
using the ChamberScope
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Figure 6. Stage pulled out of chamber. Sample holder fits into circular hole in stage.
Make sure the sample holder is fully seated.
Figure 7. Make sure that the Z position of the stage is at the exchange position after
seating the sample before closing chamber.
• Confirm that the Z-stage control did not move and is set to the exchange
position (EX), see Figure 7 above.
• Close the Chamber and push the EVAC/AIR button to evacuate the system
o Hold the chamber door securely shut until the vacuum system starts
pumping.
o Evacuation typically takes on the order of 3-5 minutes.
o The vacuum gauge is on top of the Specimen Current Meter
o There are two vacuum gauge outputs displayed on the gauge
• One is a Thermocouple gauge, which is the quarter-circle in the
upper left corner of the gauge display. The thermocouple gauge’s
range is from atmosphere to 1*10-3 torr.
• The other gauge display is a cold cathode gauge and will appear as
a number in the main gauge display. The cc gauge will not be on
until the vacuum is better than 1*10-3 torr.
o When the vacuum gauge displays 9*10-5 Torr, the system is ready for use
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Initial Startup
• Turn on HV
o The filament will turn on and be set to the last conditions used
o Set the desired accelerating voltage
Method 1
• Click on E.Beam to open the E.Beam window
• Click on Acc.Voltage
• Select the desired accelerating voltage
Method 2
• Click on either the up or down arrows next to the HV
button to reach the desired accelerating voltage
• Monitor the current accelerating voltage at the bottom of
the SEM screen
o Set the desired beam current
Click on E.Beam to open the E.Beam window
Click on Beam Curr. to open the Beam Curr. window
Set the beam current by one of the following methods
• Multi-function knobs
o X = beam current adjust
o Y = fine beam current adjust
• Clicking on the up or down arrows
• Click and drag the slider
Coarse current adjust is used for most adjustments
Fine current adjust is typically only used for reaching a precise
current level as measured via a Faraday Cup and current meter
• Confirm that the apertures are removed
• Set the magnification to the minimum
• Select the signal monitor “S”
o Bring up the brightness until the signal line just moves
The signal line is the one that extend the entire length of the screen
o Bring up the contrast until a signal envelope is observed
• Select a scan (TV1 or 2 or Scan1, 2, 3, or 4 or the reduced area scan) to obtain an
image.
• Adjust the focus to obtain an image of the sample. It may be necessary to readjust
the brightness and contrast.
o Note that the contrast and brightness should be minimized before shutting
down the instrument and should be low upon start up
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Figure 8. Objective Aperture. To insert the aperture, rotate the barrel clockwise to the
stop. The vacuum will pull the aperture in. Each stop in the clockwise direction inserts
the aperture strip one position. Aperture positions are 0 (no aperture), 1 (150um), 2
(80um), 3 (50um) and, 4 (30um).
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Final Focus
• Use the Focus knob to obtain the best possible image
o To determine if the image is in focus it is important to adjust past optimum
and then come back to optimum focus. Usually several iterations are
needed to reach optimum focus.
o If any stretching of features are observed, then astigmatism needs to be
removed from the beam
• Use the Brightness and Contrast knobs to adjust the image levels
o Adjust the brightness and contrast using the signal monitor “S”
o Best results are achieved if the signal observed just exceeds the top and
bottom signal adjustment lines
• Use the STIGMATOR knobs to remove any astigmatism from the beam
o If any stretching of features are observed this will be required
o First focus to the point that no stretching is observed
o Then adjust the X STIGMATOR knob to bring out details and improve the
apparent focus in the image
o Adjust the focus again to insure that no stretching is observed
o Adjust the Y STIGMATOR knob to bring out details and improve the
apparent focus in the image
o Adjust the focus to insure optimum focusing conditions
o Usually, this process has to be repeated several times to reach optimum
conditions
Stigmator adjustment is critical to obtaining high quality images! The astigmatism
correction should be checked each time the instrument is focused!
Data Collection
• Use the Revolution software to collect digital micrographs and X-ray data
o For more information on Revolution see the 4Pi Revolution operating
instructions below.
• Adjust the brightness and contrast for digital micrographs using the Survey mode
in Revolution
• Collect digital micrographs using E-Image
• Collect X-ray spectra using Probe
• Collect X-ray maps using X-image
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Shut Down
• Turn off any optional modes (i.e., raster rotation)
• Turn off the beam (HV off)
• Magnification to maximum
• Brightness and contrast to minimum
• Select scan 4 (slowest scan)
• Objective aperture removed
• Detectors set to fully retracted positions
• Stage set to exchange position (Z = EX, Tilt = 0, X = 30, Y = 20)
• Vent the chamber and remove the sample, then evacuate the chamber
• Fill out the log and a work order form
Robinson Detector
This section describes the use of the Robinson Backscattered Electron Detector. The
default detector is the Everhart-Thornley Secondary Electron Detector. Other detectors
are available – see Chuck for details.
DANGER! Pay attention to the position of the Robinson Detector! The Robinson
Detector inserts UNDER the pole piece and thus can easily be damaged! Repair or
replacement is very expensive ($10,000)!!! Do not touch the detector with a sample or
any part of the sample stage! Touching the detector will damage it! Pay attention to
the position of the sample when using the Robinson Detector!
• The detector must first be inserted before use. Note the position of the detector in
figure 1. The detector should be left in the retracted position except when in use.
To insert the detector:
o Turn on the chamberscope and observe that the sample and stage are
not close to the pole piece. If in doubt, get help from Chuck or Dale or
Roberto!
o Grasp the body of detector and lift the bottom guide rail, see Figure 9.
o Slowly insert the detector until it is completely inserted
Do not let the detector slam into position!
Ease the detector into place!
The detector is inserted when the body of the detector touches the
flange
• Once inserted, the detector must be chosen as the current input of the SEM
o Select the correct signal:
Click on Display Mode to open the Display Mode window
Choose Sig. Select to open the Sig. Select window
Choose B (Full, Left)
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Figure 9. A shows the Robinson detector fully retracted. B shows the bottom rail being
lifted to allow the detector to be inserted. C shows the detector fully inserted.
IMPORTANT! Turn off the detector *before* venting the chamber. The detector
will be damaged if it is not turned off before the chamber is opened!
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Danger! Use the chamberscope whenever changing the sample height! It is very easy
to damage detectors and/or the pole piece with the sample when adjusting the height!
• Turn off the detector after use and before the chamber is vented
• Retract the detector after use
o Turn on the chamberscope to observe the position of the sample with
respect to the Robinson Detector before retracting the detector
The Robinson Detector has a filter function that can clean up images, if the user is
careful.
• The filter is a low pass filter
o The Pass setting passes all frequencies of data to the SEM
High frequency noise can get through to the image using the Pass
setting
Slow scans should reduce high frequency noise
o Each setting after Pass (500kHz, 100kHz, 10kHz), cuts off the frequency
response above the selected frequency
o Fast scanning requires high frequency response, so the image will not look
good at high scan rates if a low frequency filter is chosen, e.g., TV rate
scans with a 10kHz filter are not good
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
The Robinson Detector can be used to collect images with accelerating voltages of 5kV
or more.
Revolution is freeware and facility users are encouraged to take a copy. A folder called
“Revolution for Users” is on the computer desktop for this purpose. Documentation from
4Pi is included with the folder. Merely copy the folder with Revolution onto the
computer that it will be used with and double-click on the Revolution.exe file to start the
software. No drivers or installation is needed.
Revolution Software
Revolution can run all the time and there is no need to close Revolution at the end of a
session. If Revolution is not running, start Revolution by double-clicking on the
Revolution icon on the computer desktop. A window will appear that should indicate the
version and if the software is communicating with the electronics (DXP = 1, SEM = 1,
CCD = 0). Click on OK and the software will start.
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
other than universal mode and the instructions below assume that the user is operating in
universal mode.
If the sample current is known, this can be entered under current. The current setting is
not terribly important. The current can be measured using a Faraday cup.
The Mag option should be set to “Ask and Add.” If it is set to “Ask and Add,” then the
software will ask for a magnification level prior to collecting a digital micrograph. If the
software does not ask for a magnification level, then check the Mag. option and set it to
“Ask and Add.”
It is possible to integrate several frames together in E-Image mode. See Chuck for more
details on integration. The Hitachi S-3200N does not seem to be particularly sensitive to
if the Line Sync box is checked or not.
The Image Channel Options box of the window shows which input channel is selected,
any scaling options, and the type of data to be collected. The recommended settings are
to leave Channel 1 enabled with no Autoscaling and 16 bit – Dwell (us) selected. The 16
bit – Dwell (us) selection in the data type pull down menu means that 65k gray scale
images will be collected. The dwell time per pixel is shown in the number box
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
immediately to the right of the pull down menu for data type. The larger the number, the
longer the dwell time per pixel. More dwell time means more signal for a less noisy
image and it also means that images will take longer to collect. Typically, satisfactory
results will be obtained with a dwell time of 30us/pixel in E-Image mode. Good results
are obtained in Survey mode with a 10us/pixel dwell time. This makes setting the
brightness and contrast relatively quick.
In Survey mode, the Show Waveform box should be checked. In E-Image mode, the
Show waveform box should not be checked. It is recommended that the default save
format is left as Revolution. This saves images as Revolution special files, which is not a
problem as Revolution is freeware and can be freely distributed. If images are saved as
Revolution images, then magnification information is also saved with the image.
Measuring sizes of features becomes trivial with the measurement function (M in the tool
bar). If images are saved in formats other than Revolution, the ability to quickly and
easily make measurements may be lost.
Recommended settings for publication quality images are listed below for the E-Image
setup.
Scan sizes: 1000x800, 1500x1200, 2000x1600, or 2500x2000 pixels
Frame Average: Typically unchecked. If the sample is charging, multiple fast
frames, i.e., images taken with a low dwell time per pixel, can be averaged
together.
Line Sync: checked
Spatial Frame Lock: unchecked
Channel 1: Autoscaling – None, Enable checked, 16-bit Dwell (us), 20.
Channel 2: Autoscaling – None, Enable not checked, 16-bit Dwell (us), 1.
Access Control: Unlock All
Line Profile Option: Show line profile not checked
Default Save Format: Revolution
Save as Displayed: not checked
Digital Imaging
To take a digital image, first set the brightness and contrast using Survey mode. Click on
Survey mode and an image will appear at a relatively fast scan rate (controlled by the
scan size and dwell time in the setup window) with a signal monitor shown in red in the
center of the screen. Adjust the brightness and contrast such that all of the desired signal
can be observed with the signal monitor. Generally, this will mean that the signal will
extend almost all the way from 0 (bottom dashed red line) to 100% (top dashed red line).
It is generally smart to turn the contrast down until the signal line is flat, set the flat line
to zero, and then increase the contrast until the signal is satisfactory. Note that the image
should be aesthetically pleasing no matter what the signal monitor suggests!
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Once the brightness and contrast are set, collect a digital image by clicking on the E-
image button. A pop-up window will ask for the magnification. Enter the magnification
from the SEM and either press the enter button on the keyboard or click on OK. An
image will form in a new window. If the user is not prompted to enter a magnification,
select “Ask and Add” from the Mag pull down dialog box.
The image can be saved as either a Revolution file (recommended), a TIFF, a JPEG, or a
BMP file.
X-Ray Analysis
For information on understanding and interpreting X-ray spectra, please refer to
J.Goldstein, et. al., Scanning Electron Microscopy and X-ray Microanalysis, New York,
Plenum, 2003. For more information on how the 4Pi Universal Spectral Engine handles
X-ray analysis please refer to the 4Pi documentation.
It is a good idea to have an overvoltage of at least a factor of two above the maximum X-
ray energy that is to be collected, e.g., for a 8.04 keV Cu Kalpha X-ray, one would want
the accelerating voltage to be at least 16kV with 20kV being a better choice. See
Goldstein, et. al., for more information on overvoltage. It is important to set the
accelerating voltage to that being used by the instrument in the kV window of the
Revolution toolbar.
In general, the working distance should be set to 17mm +/- 2mm to collect X-ray data. It
is best to move the X-ray detector in to the bottom stop, which will maximize the solid
angle of collection and enhance efficiency.
To collect X-ray spectra, there are several methods with two being the most common.
One is to zoom on the region of interest and collect a spectrum of the area, the other is to
take an image and select points from which to collect X-ray data.
Method 1. Focus on the region of interest (ROI) such that the ROI fills the entire field of
view of the SEM. Click on Probe in the Revolution toolbar. An X-ray spectrum of the
ROI will appear.
Method 2. Collect a digital image. Click on the “Rectangle” button and draw a box
around the ROI. Then right click on the box and choose to “EDX selected area.” An X-
ray spectra of the ROI will be collected in a new EDS spectrum window. It is possible to
select a single point with the point button. It is also possible to queue multiple areas to
collect X-ray spectra by creating multiple ROIs and then right click on the image and
choose “Queue EDX items.” Spectra will be created in the order in which the ROIs were
placed on the image.
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
Multiple spectra can be collected in a single spectrum file. This allows for easy data
management since all the spectra for a single sample can be collected in a single file. If
an EDS window is open, any new spectra will be collected in the open spectrum file. If a
new window is desired, click on File -> New Catalog to open a new EDS window. If
multiple EDS windows are open, the new spectrum will be collected in the active
window, which will be the EDS window on top.
Once a spectrum is collected, X-ray peaks can be marked by right clicking on the
spectrum and choosing to “Open KLMs.” This will open the X-ray Spectra KLMs
window, which allows the user to mark the location of specific elemental X-rays. If the
X-ray line is known, the user can simply click on the element to mark its peaks. If the
sample is unknown, the user can use the right-left arrow keys on the keyboard to scroll
through the elements until the lines that correspond to measured peaks are observed.
For more information on correctly identifying X-ray peaks, see the Goldstein et. al.
reference or see Chuck or Dale or Roberto. For more advanced techniques, such as
Quantitative analysis, X-ray mapping or Linescans, please see Chuck or Dale or Roberto.
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Analytical Instrumentation Facility
2410 Campus Shore Drive
318 Larry K. Montieth Research Center
www.ncsu.edu/aif
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