Transmission Electron Microscopy:

1
Transmission Electron Microscopy What
information can be obtained from the
ultrathin section?
Anna Klintsova
2
Beckman Institute TEM facility

• TEM
• Tissue processor
• Ultracut
• Carbon evaporator
• Dark rooms
• People who can help, consult and advise

3
How image is created in light and transmission
electron microscope
4
One of the first EM images of the cell
fibroblast with nucleus and mitochondria
5
Resolution of Philips CM200 microscope distance
between lines 2 .3 A. Image Au on carbon
6
Specimen is desiccated, suffocated, electrocuted,
cooked and nuked
7
Lynx Tissue Processor - great time- and
health-saver
8
Reichert Ultracut S - latest model, allows to
cut sections 10-100 nm thick
9
Ultrathin (50-70 nm) sections are collected on
the formvar-coated grid
10
Excitatory synapse in the visual cortex of rat
Vesicle size - 50-100 nm
11
Negative staining adenoviruses
100 nm
12
Vacuum Evaporator evaporates carbon, and metals
too!
13
Three-dimensional world - on two-dimensional
sections
14
Microvillii in the rat intestine Horizontal
section
15
Microvillii in the rat intestine Vertical section
16
Stereology - unbiased method of quantitative
analysis of three-dimensional structures on
sections
17
Cycloid grid - method of fast and unbiased
estimate of surface density and surface area
18
Digitally colored TEM image of macrophage
19
Common protozoan parasite that causes
toxoplasmosis.
20
Ultramicrotomed Supramolecular Film
78 Å
ultramicrotomy reveals the same layer spacing as
SAXS
Intensity (au)
39 Å
1.0
2.0
3.0
2???????
79 Å
Fourier filtering reveals molecular layer
information
Cross section of a 300 nm film stained with OsO4
Courtesy of Martin Pralle
21
Direct Imaging of Polar Order
repeat unit 79 Å
unstained
stained
crystalline
stained
control experiment verified PPV staining by OsO4
supramolecular film
Courtesy of Martin Pralle
22
CdS
polymer substrate
reactive gas (H2S)
lyotropic liquid crystal directly templates the
growing CdS film as H2S diffuses through the
polymer substrate
columns of hexagonal liquid crystal perpendicular
to substrate
Courtesy of Paul Braun
23
Top
Bottom
holey carbon
TEM cross-section of CdS film grown with 15
minute H2S exposure
Courtesy of Paul Braun
24
zoom in of cross-section, channels run
perpendicular to substrate
Courtesy of Paul Braun
25
Immunocytochemistry - localization of
antigen/antibody reaction on ultrastructural
level