Webinar Series in TEM: TEM Practical Session

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The world leader in serving science

Riza Iskandar

Webinar Series in TEM:

TEM Practical Session

Application Scientist Material Science

Material and Structural Analysis - APAC

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3,834.07 km

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TEST QUIZ

Which of the following is a figure of TEM?

A B C

D

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TEST QUIZ

Which one is the image taken from TEM?

A B C

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TEM Practical Session: Talos F200X

Outline

• Introduction

• Talos Tour

• Sample Information

• Nanoparticles

• STO FIB Lamella

• Case 1: Nanoparticles

• TEM-BF

• TEM-DF

• SAED

• HRTEM

• STEM 4 Detectors

• STEM-EDS

• MAPS

• Case 2: STO FIB Lamella

• Smart Tilt

• HRTEM

• HRSTEM-EDS Map

• EDS Mapping

• Case 3: Life Science*

•

Visualizing Proteins and Macromolecular

Complexes by Negative Stain

•

Cryo-Electron

Tomography

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Introduction

• Fully remotely controlled and in a Box

• Automation:

• Daily Tuning: Focus, Z, C2 aperture, pivot points

• CrsytalPack: Automatic tilt to a present zone axis

• (S)TEM MAPS (Google Maps on your sample)

• Automated EDS tomography

• Live images of all elements of the periodic table

• Simultaneously imaging of light and heavy elements

• Suitable for energy application materials, such as LiB materials

• Largest in-situ application space

• Live chemical reaction for electrolyte materials Talos F200X

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Introduction

Talos F200X: Visual Tour

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Introduction

Sample Handling

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Introduction

Loading Sample

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Introduction

Inserting Holder - 1

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Introduction

Inserting Holder - 2

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Introduction

Preparing the Experiment

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TEM Practical Session: Talos F200X

Outline

• Introduction

• Talos Tour

• Nanoparticles

• STO FIB Lamella

• TEM-BF

• TEM-DF

• SAED

• HRTEM

• STEM-EDS

• MAPS

• Smart Tilt

• HRTEM

• HRSTEM-EDS Map

• EDS Mapping

• Case 3: Life Science*

•

Visualizing Proteins and Macromolecular

Complexes by Negative Stain

•

Cryo-Electron

Tomography

(14)

Sample Information

• A mix of various metals particles (Fe, Ni, Cu, Zn, Ag)

• Prepared by conventional powder preparation

• Nanoparticles disperse on a TEM grid coated by carbon thin film

• Particles are pretty large for TEM analysis

• Carbon thin film increase the sample thickness and introduce contamination during STEM-EDS analysis

Nanoparticles

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Sample Information

• Cubic Perovskite SrTiO₃ <100>

• Prepared by Focused Ion Beam Technique STO FIB Lamella

TEM Grid for FIB lamella

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TEM Practical Session: Talos F200X

Quiz

1. Which aperture that is required to acquire TEM-BF image

a. Condenser aperture

b. SAED aperture

c. Objective aperture

2. Which aperture that is required to acquire TEM-DF image

b. SAED aperture

3. Which aperture that is required to acquire selective area electron diffraction pattern

b. SAED aperture

c. Objective aperture

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TEM Practical Session: Talos F200X

Quiz

4. To perform a good and reliable TEM imaging, which steps should be done:

a. Bring the sample into eucentric height

b. Always use objective aperture to acquire TEM-BF image

c. Always use selective area aperture to acquire SAED pattern

d. Always use one specific camera length when acquire SAED pattern

e. Condenser aperture must be centered 5. When we need to set the eucentric height

a. At the beginning of the session

b. When we move the sample

c. When we switch between image to diffraction mode

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TEM Practical Session: Talos F200X

Outline

• Introduction

• Talos Tour

• Nanoparticles

• STO FIB Lamella

• TEM-BF

• TEM-DF

• SAED

• HRTEM

• STEM-EDS

• MAPS

• Smart Tilt

• HRTEM

• HRSTEM-EDS Map

• EDS Mapping

• Case 3: Life Science*

•

Visualizing Proteins and Macromolecular

Complexes by Negative Stain

•

Cryo-Electron

(19)

Eucentric Height

• Start at low

magnification  10000x

• Use mechanical wobbler to tilt the sample +/- 15 on α

• Use Z to move the sample into the

eucentric height, i.e., fewer movements

• Change to higher magnification  350kX

• Use Z again to remove Fresnel fringes

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TEM Bright Field

• Select area of interest

• Switch to diffraction

• Insert and adjust the position of the objective aperture

• Switch back to image mode and acquire the image

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TEM Dark Field

• Switch to diffraction with no SAED aperture

• Insert and centering the smallest objective aperture

• Move the reflected spot to the center using multifunction knobs

• Switch to image mode to show TEM- DF

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Electron Diffraction - SAED

• Insert SAED aperture

• Switch to diffraction

• Bring the sample into a specific zone axis

• Tilt on alpha and beta axis

• Move the primary beam to the center

• Make the beam mostly parallel

• Turn intensity knob clockwise until specific C2 lens value

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STEM 4 Detectors

Panther Detector

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STEM-EDS

• For a single EDS detector, tilt the sample towards to EDS detector

• Draw an ROI marking

• Check the EDS acquisition parameter

• Start EDS acquisition

• Cross-correlation images will be shown in Velox acquisition

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MAPS STEM

• It can be performed both in TEM and STEM

• Start MAPS software

• Input all necessary parameters

• Run the process

• The results will show both STEM HAADF image and EDX elemental maps

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MAPS TEM

• It can be performed both in TEM and STEM

• Start MAPS software

• Input all necessary parameters

• Run the process

• The results will be the final image

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MAPS for Corelative Imaging

• Link of different imaging techniques:

Optical Microscope

SEM

TEM

STEM

Tomography

Correlative Imaging of Optical Microscope, SEM, TEM, STEM and Tomography for Carbon Nanotube

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TEM Practical Session: Talos F200X

Quiz

1. Which aperture that is required to acquire selective area electron diffraction pattern

a. SAED aperture

b. Objective aperture

c. Condenser aperture

2. Which aperture that is required to acquire TEM-DF image

b. Objective aperture

c. SAED aperture

3. Which aperture that is required to acquire TEM-BF image

b. SAED aperture

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TEM Practical Session: Talos F200X

Quiz

4. To perform a good and reliable TEM imaging, which steps should be done:

a. Bring the sample into eucentric height

b. Always use objective aperture to acquire TEM-BF image

c. Always use selective area aperture to acquire SAED pattern

d. Always use one specific camera length when acquire SAED pattern

e. Condenser aperture must be centered 5. When we need to set the eucentric height

a. Only at the beginning of the session

b. When we move the sample

c. Only when we switch between image to diffraction mode

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TEM Practical Session: Talos F200X

Quiz

6. When acquiring HRTEM images, following statements are true except:

a. Sample must be brought at eucentric height

b. Defocused must be controlled in the interval of -100 ≤ f ≤ +100 (nm)

c. Objective aperture must be inserted

d. Sample must be oriented at specific zone axis 7. Which is from the following statements is true:

a. STEM resolution depends on the probe size

b. STEM detectors can not be user simultaneously

c. Focusing image on STEM is more difficult than in TEM mode

d. STEM can only be used to analysis nanoparticle

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TEM Practical Session: Talos F200X

Quiz

8. Following conditions are required to acquire a good STEM images:

a. Small C2 aperture

b. Long acquisition time to get high signal to background ratio

c. Smallest spot size

d. Adjust the brightness and contrast 9. A good EDS results depend on:

a. High counts

b. Long acquisition time

c. Large C2 aperture

d. Thin sample

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TEM Practical Session: Talos F200X

Outline

• Introduction

• Talos Tour

• Nanoparticles

• STO FIB Lamella

• TEM-BF

• TEM-DF

• SAED

• HRTEM

• STEM-EDS

• MAPS

• Smart Tilt

• HRTEM

• HRSTEM-EDS Map

• EDS Mapping

• Case 3: Life Science*

•

Visualizing Proteins and Macromolecular

Complexes by Negative Stain

•

Cryo-Electron

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Smart Tilt

1

2

3

4

• Activate Smart Tilt function

• Under Stage tab

• Click Display

• Make sure “Use compucentric tilting” is selected

• On the smart Tilt

window, click Zone axis Tilt (the first icon on the left)

• Follow the instruction

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High-resolution TEM

• 2D atomic arrangement

• Sample oriented on

specific crystal orientation

• Low index zone axis

• Go to higher magnification

• Beware with the

movement of the area of interest

• Re-check eucentric height

• Fine-tuning of focus

• Intensity to clockwise, focus to counter-

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HRSTEM STO

• Bring the sample into the low zone axis

• Re-focus the image with Z

• Increase magnification up to 5 Mx

• Re-focus the image with a focus window

• Correct astigmatism

• Turn off the focus window

• Start to acquire the image

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HRSTEM-EDS STO

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TEM Practical Session: Talos F200X

Quiz

1. To acquire high-resolution STEM image:

a. The sample must be oriented at low zone axis

b. The sample must be in eucentric height

c. Minimal magnification is 5.000.000x

d. Refocus and correct astigmatism

2. Reducing contamination can be done by:

a. Clean the sample using plasma gas prior inserting into microscope

b. Heat up the sample prior inserting into microscope

c. Tilting the sample to specific angle before acquiring the image

d. Using cool trap for trapping the gas molecules on sample surfaces

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TEM Practical Session: Talos F200X

Quiz

3. Acquiring HRSTEM-EDS elemental map requires:

a. At least dual EDS detectors

b. Drift correction is turned on

c. Clean sample area

d. Large enough beam current

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TEM Practical Session: Talos F200X

Outline

• Introduction

• Talos Tour

• Nanoparticles

• STO FIB Lamella

• TEM-BF

• TEM-DF

• SAED

• HRTEM

• STEM-EDS

• MAPS

• Smart Tilt

• HRTEM

• HRSTEM-EDS Map

• EDS Mapping

• Case 3: Life Science*

•

Visualizing Proteins and Macromolecular

Complexes by Negative Stain

•

Cryo-Electron

Tomography

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TEM Practical Session: Life Science*

Visualizing Proteins and Macromolecular Complexes by Negative Stain

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TEM Practical Session: Life Science*

Cryo-Electron Tomography

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TEM Practical Session: Life Science*

Quiz

1. Why is the maximum acceleration voltage for life science TEM experiments limited to 120kV?

a. To enhance the contrast of biological sample

b. To improve spatial resolution

c. To reduce beam damage to the sample due to electron’s energy

d. To reduce contamination

2. What is the function of carbon coatings on TEM grids coated with plastic film?

a. To enhance conductivity

b. To increase the electrostatic force that keeps the sample attached to the grid

c. To mark the side of the grid that used to put the sample

d. To reduce the charging effect

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TEM Practical Session: Life Science*

Quiz

3. Why do virus samples need to be cooled by liquid Nitrogen?

a. To reduce contamination during the experiment

b. To enhance the spatial resolution

c. To keep the structure of the viruses as TEM operated at high vacuum

d. To prevent the operator is being infected by viruses

4. Type of TEM that can be used to analyze the virus structure up to subs-angstrom by using cryo- tomography:

a. Titan Krios

b. Talos F200X

c. Talos L120C

d. Tecnai

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TEM Practical Session: Talos F200X

Summary

• TEM operation is more complex than any other techniques

• Nowadays, the operation is more user-friendly and full automated

• A stable and powerful TEM is necessary to acquire different information

• High brightness electron gun, highest spatial resolution, high counts EDS signal

• Remote Operation TEM

• Borderless TEM operation: remotely operation open possibility for more extensive collaboration

• The best solution for a large and centralized research center

• The best solution in a pandemic situation: less traveling, best practice for social distancing

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Webinar Series in TEM: TEM Practical Session