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TEM Sample Prep and Optical Microscopy

TEM Sample Prep and Optical MicroscopyOpen original PDFFull searchable text · 44 pagesRefer to the original PDF for tables, figures and page layout.Page 1 Page 2 Introduction • Materials Engineer • Allied High Tech Products, Inc. • 4 years of laboratory experience • Primary responsibilities: • Procedure development for various materials • Image Analysis • Customer support/training • Allied High Tech Products, Inc. • Founded in 1983 • An American Manufacturer and distributor of high quality…

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Manufacturer / publisherAllied High Tech Products
LanguageEnglish
Captured2026-10-17
Document typedownloads
PDF pages44
CategoryMetrology & quality control
Updated2026-10-17T21:12:06.592434+00:00
Allied High Tech published content
Source: https://www.alliedhightech.com/Media/Default/EquipmentBrochures/TEM%20Sample%20Prep%20and%20Optical%20Microscopy.pdf. Manufacturer claims and terminology are reproduced as source material.Explore Brochures & catalogues ↗

TEM Sample Prep and Optical Microscopy

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Full searchable text · 44 pages

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Page 1


Page 2
Introduction
•
Materials Engineer
•
Allied High Tech Products, Inc.
•
4 years of laboratory experience
•
Primary responsibilities:
•
Procedure development for 
various materials
•
Image Analysis 
•
Customer support/training
•
Allied High Tech Products, Inc.
•
Founded in 1983
•
An American Manufacturer and 
distributor of high quality  
equipment and consumables for 
metallographic sample 
preparation and analysis.
•
Zeiss Optical Microscope National 
Dealer 
•
Mitutoyo Rockwell and MHT 
Dealer
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 3
Topics
• Basics
• What is a TEM sample?
• Requirements?
• Methods of preparing TEM 
samples?
• Mechanical preparation of a TEM 
sample
• Preparation process
• The role of optical microscopy in 
the sample preparation process
• Stopping point: Pre-FIB thinning 
or TEM Wedge?
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 4
TEM – Transmission electron 
microscopy is a microscope technique in 
which a beam of electrons is 
transmitted through a specimen to form 
an image.
TEM Sample – Specimen that is usually 
ultra thin (<100 nm) to allow electrons 
to transmit through.
What is a TEM Sample?
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A
[3] Zeiss HRTEM


Page 5
TEM Sample Requirements
TEM samples must be electron transparent and generally the goal is a sample 
thickness of <100 nm
Any deformation from thinning process or previous processing must be removed
Small area, < 3 mm or less, to fit various TEM grids and holders
© A L L I E D  H I G H  T E C H  P R O D U C T S
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[3] Specimen support grids of different 
mesh size and shape
[3] Two-specimen holder with double-tilt


Page 6
Common Methods of Preparation
•
Ion Milling/FIB – Focused Ion Beam
•
Process that involves bombarding a TEM 
sample with energetic ions or neutral atoms 
and sputtering material from the sample 
until it is thin enough to study in the TEM 
[3].
•
The most versatile thinning process that can 
be used for a wide variety of materials, 
however very expensive to initially invest in 
and to run.
•
Disadvantage is implantation of source 
material and amorphous layer is created.
•
Most commonly used in semiconductors. 
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A
[3] Schematic of a two-beam FIB and stages in 
making TEM samples using a FIB


Page 7
Common Methods of Preparation
•
Electropolishing
•
Sample is immersed in an electrolyte and 
then subjected to a direct electrical current. 
The sample is maintained anodic, with the 
cathodic connection being made to a nearby 
metal conductor. [2]
•
Anodic dissolution of sample creates 
polished surface.
•
Relatively quick and can produce samples 
with no mechanical damage, however can 
only be used for electrically conducting 
metals and alloys.
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A
[3] The Window Method


Page 8
Common Methods of Preparation
•
Mechanical Preparation: Can be 
performed by hand or with the aid of a 
machine
•
Manual: Utilizes hand tools (Tripods) that 
allow the user to make angle adjustments
•
Semi-Automatic: Precision polishers, such 
as the MultiPrep™ System, with digital 
indicators and dual micrometers allow: 
– Monitor amount of material removed in real 
time in 1-micron increments
– Precise sample tilt adjustments relative to 
the abrasive plane. Alignment ensures 
controlled angles.
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 9
Mechanical Preparation
• Advantages
– TEM Wedge capabilities
– All damage removed using the 
3x rule
– No foreign material implanted
– With experience it is repeatable 
and fast 
– Lower cost of required 
equipment
– Decrease milling time and costs
• Disadvantages
– Requires careful handling 
throughout process
– Greater learning curve than others 
techniques
– Some samples may require one of 
the other TEM preparation 
methods such as metals due to 
internal strain
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 10
Preparation
• Area of interest needs to be 
identified; site-specific or bulk?
• Once determined, sample needs 
to be prepared
• Preparing TEM samples is 
essentially performing two 
cross-sections
First side 
cross-section
Second side 
cross-section
© A L L I E D  H I G H  T E C H  P R O D U C T S
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Target


Page 11
Thinning First Side
• Sample placed onto a          
cross-sectioning paddle       
using mounting wax
• Paddle placed on hot plate to 
melt wax
• Light pressure can be applied 
using a cotton-tipped applicator 
to assist in parallel registration 
of the sample to the fixture.
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 12
Thinning First Side
• Stereomicroscope can greatly 
assist in aligning the sample to 
the fixture
• Tools such as the MultiPrep™ 
may have methods of correcting 
for the angle; however, using a 
stereomicroscope can minimize 
the amount of corrections 
required
To paddle 
edge
Align sample 
features
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 13
Thinning First Side
• While the paddle is hot, it 
can be taken to a 
stereomicroscope
• This allows for sample 
adjustments before the 
wax cools and hardens
Initial adjustments under microscope
Use tweezers/tool to 
make adjustments
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 14
Thinning First Side
• Lapping films used as 
abrasive
– Provide excellent edge 
retention and maintain 
coplanarity
– Typically used for 
unencapsulated cross-
sectioning, TEM preparation, 
backside polishing, etc.
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 15
Thinning First Side
• Initial grinding with a coarse 
lapping film, such as 30 µm, will 
give a ground edge
• This ground edge can be used to 
obtain angle information using 
optical microscopy
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 16
Thinning First Side
• Compound light microscopes 
will give the resolution and 
details needed to perform 
corrections
• Imaging software can be used to 
obtain angle measurements
88.87°
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 17
Thinning First Side
• Angle corrections can be made 
using the micrometers
• Each tick mark is 0.02°
• More material can be removed to 
obtain a new ground edge
• With the new ground edge, the 
process can be repeated until an 
angle correction is no longer 
necessary
0.02°
Corrected alignment
Previous alignment
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 18
Target: Site-Specific or Bulk?
Site-Specific
• Particular feature on 
sample
• Failure Analysis
Must stop at target, without 
damage from previous steps
Bulk
• Homogeneous material
Can stop anywhere, however 
ensure no damage from 
previous steps
© A L L I E D  H I G H  T E C H  P R O D U C T S
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Target


Page 19
3X Rule
•
Damage is introduced into a sample 
throughout the preparation process. 
•
This damage is not only represented by a 
scratch pattern on the surface, but also by 
smaller micro-cracks that can propagate 
further into the sample that may not be 
seen by eye or optical microscopes          
(A 30 µm lapping film will not cause only
30 µm deep scratches).
Sample
Damage
(micro-
cracks)
30 µm DLF
15 µm DLF
AOI
9 µm DLF
30 µm 
DLF
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 20
3X Rule
•
If a 30 µm lapping film is used and it is followed by a 15 µm lapping film step, the 15 µm DLF 
must remove at least 90 µm of material to completely remove the damage from the 30 µm 
DLF. Similarly, if a 30 µm DLF is used and it is followed by a 6 µm DLF, then the 6 µm DLF 
must remove at least 90 µm of material to completely remove the remaining damage.
3X Rule
Current Step
(DLF)
Current 
Distance from 
Target
Previous Step
Remove At 
Least 3x Prev. 
Step Size
Distance to 
Target After 
Current Step
30 µm
Vary
N/A
N/A
192 µm
15 µm
192 µm
30 µm
90 µm
102 µm
9 µm
102 µm
15 µm
45 µm
57 µm
6 µm
57 µm
9 µm
27 µm
30 µm
3 µm
30 µm
6 µm
18 µm
12 µm
1 µm
12 µm
3 µm
9 µm
3µm
0.5 µm
3 µm
1 µm
3 µm
At Target
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 21
3X Rule
•
Other materials, such as ductile steels, 
may only require a “2X Rule”, while 
fragile, brittle ceramics, where cracks 
can propagate further into the sample, 
may require a “4X Rule”.
•
The main concept to understand is that 
no matter what steps follow, all damage 
introduced by the previous abrasive 
must be removed to get an accurate 
representation on the microstructure.
Sample
Damage
(micro-
cracks)
30 µm DLF
15 µm DLF
AOI
9 µm DLF
30 µm 
DLF
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 22
Flipping Sample
• 2nd side needs to be prepared to 
thin sample
• Previous method using cross-
sectioning paddle can’t be used; 
can’t overhang sample properly
• TEM paddles with Pyrex® inserts 
are used to continue with the 2nd
side
Polished edge
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 23
Flipping Sample
• Surface of Pyrex® requires grinding
– Grinding the surface of the Pyrex® will align it with that of the platen, 
ensuring parallelism
• Pyrex® can be ground using diamond lapping films
– 6-3 µm finish is good enough
– Do not want to polish the surface as having some scratches will help 
with adhesion
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 24
Site-Specific Orientation
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 25
Site-Specific Orientation
• With site-specific 
orientation, the area of 
interest is parallel to the 
Pyrex® edge
Area of Interest
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 26
Bulk Orientation
© A L L I E D  H I G H  T E C H  P R O D U C T S
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Page 27
Bulk Orientation
• With bulk orientation, the 
area of interest is 
perpendicular to the 
Pyrex® edge
Area of Interest
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 28
Flipping Sample
•
Sample waxed onto a glass slide
•
~1 mm thick piece sectioned from sample
– This allows placement onto TEM paddle without risk of the sample breaking 
off due to being too tall
•
Sample should be cleaned to remove abrasive particulates, excess wax, 
etc.
1 mm
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 29
Flipping Sample
• To ensure stability, a thin, 
uniform layer of wax needs to 
be used.
• Wax Bead Method
– Place sample, polished side down, 
into wax bead
– Pick up sample and move over to 
empty part of glass slide
– Allow wax to wick away
– Repeat process 2-3 more times to 
end up with thin wax layer
© A L L I E D  H I G H  T E C H  P R O D U C T S
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Page 30
Flipping Sample
• Polished side placed face down 
onto the Pyrex®
• Positioned near the edge to 
allow easier thickness 
measurements later
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 31
Flipping Sample
• Stereomicroscope is great for 
aligning to the edge of the 
Pyrex®
Tweezers used to make 
slight adjustments
Sample edge flush with 
Pyrex® edge
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 32
Flipping Sample
• Stereomicroscope is great for 
aligning to the edge of the 
Pyrex®
Tweezers used to make 
slight adjustments
Sample edge flush with 
Pyrex® edge
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 33
Second Side
•
Second side, whether site specific or not, now has a final target
– Can lose sample if thin/polish for too long
•
Checking sample thickness becomes extremely important if you want 
to remove all the damage from previous steps
•
Certain machines, such as the MultiPrep™ System, may have 
indicators or other methods to track material removal
– However, checking progress frequently with an optical microscope is a good way 
to verify material being removed: trust but verify
1018 µm
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 34
Second Side
• Mounting the sample close to 
the edge allows for proper 
focusing on the sample to 
measure side 1 edge
• Compound scope can give 
required resolution to view 
sample thickness and make 
measurements
Pyrex®
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 35
Goal: Pre-FIB or TEM Wedge?
Pre-FIB Thinning
•
Site-Specific Orientation
•
Remove the sample to prepare 
for FIB thinning
•
Sample can be removed by 
placing paddle into container 
with acetone (wax dissolves in 
acetone)
•
Filter paper placed around 
paddle so sample will eventually 
pop-off and land on paper
TEM Wedge
•
Bulk Orientation
•
Induce an angle to create a wedge 
sample
•
Angle induced will vary depending 
on the material being prepared
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A
Thinned to electron 
transparency
Thick enough for 
handling


Page 36
Inducing Wedge
• Micrometers on MultiPrep™ used 
to induce an angle on the sample 
to create a wedge
• Wedge allows for a sample thinned 
on one edge, with a thick enough 
edge for handling
Parallel contact plane 
0.02°
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A
Thinned to electron 
transparency
Thick enough for 
handling


Page 37
Wedge Polishing
•
Certain TEM samples, along with 
the use of a transmitted light 
microscope, can display a series of 
colors in regions less than 10 µm 
thick
•
Fringes can also occur in regions 
less than 2 µm thick
•
The thickness that color 
corresponds to will vary depending 
on materials; however, some are 
well documented, such as silicon
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 38
Wedge Polishing
Color and fringe information can be 
used to assist in the preparation of 
TEM samples, as they are a guide to 
overall progress during a TEM wedge 
procedure 
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A
[4] Correspondence between Si wafer thickness and color of transmitted light


Page 39
Wedge Polishing
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 40
Wedge Polishing
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 41
Wedge Polishing
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 42
Wedge Polishing
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 43
•
1. Allied High Tech Products, Inc., http://www.alliedhightech.com/. Accessed 23 May 2018.
•
2. Delstar Metal Finishing, Inc., Electropolishing A User’s Guide to Applications, Quality Standards 
and Specifications, https://www.delstar.com/assets/pdf/epusersguide.pdf. Accessed 23 May 2018.
•
3. Williams, David B, and C B. Carter. Transmission Electron Microscopy: A Textbook for Materials 
Science. New York: Plenum Press, 1996. Print.
•
4. Yougui Liao. Practical Electron Microscopy and Database. URL: 
http://www.globalsino.com/EM/page2805.html. GlobalSino 2007.
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A


Page 44
Pablo Mendoza
Materials Engineer, Technical Services
Allied High Tech Products, Inc.
pmendoza@alliedhightech.com
© A L L I E D  H I G H  T E C H  P R O D U C T S
P A B L O  M E N D O Z A

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