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www.reading.ac.uk/EMLab Introduction to the Electron Microscopy Laboratory Peter Harris

Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

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Page 1: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

www.reading.ac.uk/EMLab

Introduction to the Electron Microscopy Laboratory

Peter Harris

Page 2: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Who we are

Why electron microscopy?

Specimen preparation for SEM; advanced forms of SEM

EMLab’s facilities

Specimen preparation for TEM

Electron diffraction and X-ray analysis

Becoming an EMLab user

Tour/questions

Electron Microscopy Laboratory

Page 3: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Electron Microscopy Laboratory

School of Chemistry, Food and Pharmacy (SCFP)

Chemical Analysis Facility

Electron Microscopy Laboratory

Page 4: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Electron Microscopy Laboratory

Pharmacy: SEM of polyurethane microcapsules

Chemistry: TEM of peptide nanotapes

Food Biosciences: SEM of bacteria

Biological Sciences: SEM of flower from insectivorous plant

Soil Science: SEM of biochar

Page 5: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Microscopy basics: why electron microscopy?

The highest resolution achievable by a lens is given by the Rayleigh criterion:

sin

.

nd

610

where d is the resolution, λ is the wavelength of emitted light, and n is the refractive index of

the lens and α is the semi-angle subtended by the aperture.

Wavelength of an electron (200 kV): 0.0025 nm.

Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

Highest resolution of scanning electron microscope ~ 10 nm.

Wavelength of light (blue): ~ 400 nm.

Highest resolution of optical microscope ~ 200 nm.

Page 6: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Electron Microscopy Laboratory

Scanning electron microscope

Electron gun

Specimen

Electron gun

Condenser lens

Objective lens

Specimen

Projector lens

Fluorescent screen

Electron gun

Digital camera

Condenser lens

Objective lens

Specimen

Projector lens

Fluorescent screen

Electron gun

Digital camera

Transmission electron microscope

Workshop: November 5 Workshop: November 12

Page 7: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Electron Microscopy Laboratory

EMLab’s scanning electron microscopes

Cambridge S360 High Vac SEM FEI Quanta Environmental SEM

Page 8: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Electron Microscopy Laboratory

EMLab’s transmission electron microscopes

FEI/Philips CM20 Analytical TEM JEOL 2010 High resolution TEM

Page 9: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Looking at “wet” samples in the high vacuum SEM

Animal and plant tissues up to 98% water.

Options for examining these materials in the high vacuum SEM:

Freeze drying

Chemical fixation, critical point drying

Page 10: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Critical point drying

Substitution of water with liquid CO2, which has

lower surface tension

Controlled removal of liquid CO2

Page 11: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Coating SEM specimens

For conventional (high vacuum) SEM, surface

must be electrically conducting.

Usually coat with gold, but can also coat with

chromium or carbon.

Page 12: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Sample prepared by fixation & critical point drying

Euphorbia seed. Amal Al Hasan, SBS.

Page 13: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Cryo-SEM

Workshop: November 19

Potato showing starch grains

Page 14: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Environmental SEM

Workshop: November 19

Cooled specimen

• Special pumping system allows water vapour to be introduced into chamber.

• Cooled specimen to retain moisture.

• New (backscattered) electron detector.

Page 15: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Environmental SEM

• Raspberry buds : Hiroyuki Imanishi, Plant Sciences

Page 16: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Specimen preparation for TEM

Specimens for TEM

3 mm TEM grid

Carbon films

Page 17: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Specimen preparation for TEM

Inorganic powders – easy!

Page 18: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

TEM in physical science

Platinum/alumina catalyst

Gold nanoparticle

Carbon nanostructures

Page 19: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

TEM in biology

Adenovirus negatively stained with

uranyl acetate

Page 20: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Specimen preparation for TEM

Biological TEM – preparing specimens from tissue

Microtome

Fix dehydrate set in resin section deposit on grids stain

Page 21: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Specimen preparation for TEM

Specimen preparation laboratory

Page 22: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Specimen preparation for TEM

Microtome

Page 23: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Biological TEM – thin section

Kidney

TEM in biology

Page 24: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

An electron microscope is not just a microscope

Elemental analysis - determining the composition of materials

Workshop: November 26 Diffraction pattern (crystalline C60)

Electron diffraction in the TEM

Page 25: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

X-ray Analysis: Mapping

Workshop: November 27

Tracey Finn/Amy Smith (Ure Museum)

Fragments of Egyptian pot

Page 26: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Becoming an EMLab user

Sherrie Foo

Page 27: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Booking system

Page 28: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Remaining workshops

Date

Topic

5 November 2014 Scanning Electron

Microscopy

12 November 2014 Transmission Electron

Microscopy

19 November 2014 Environmental & Cryo

SEM

26 November 2014 X-ray Analysis and

Elemental Mapping

Page 29: Introduction to the Electron Microscopy Laboratory · Wavelength of an electron (200 kV): 0.0025 nm. Highest resolution of transmission electron microscope ~ 0.1 nm. Individual atoms!

EM Workshops 2014

Electron Microscopy Laboratory

Tour/questions


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