c
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Vacuum Subsystem

Electron Gun Subsystem

Electron Lens Subsystem

Sample Stage

More Electron Lenses

Viewing Screen w/scintillator

Camera Chamber
 c


 

= ilament saturation
= ilament centering
= Spot size (Condenser
Lens Current)
= Condenser aperture
 ° 

c
= Control objective lens current
=
djust astigmatism correction coils too
= se large screen at low mags
= se small screen at high mags
= eware of lingering on an area too long
= terate focus and stigmators
= Can take through-focus-series
 ocusing on a hole in a thin carbon film
( resnel ringes)

(a) underfocused objective lens (bright fringe);

(b) at focus (no fringe);
(c) overfocused objective lens (dark fringe).

Note also the change in appearance of the

carbon fine grain.
Magnification ~750,000X.

( rom
gar,p.137).
 h  


= Sort of like the SEM in that astigmatism shows up as a

directional defocus

= Must correct condenser lens, objective lens and projector

lens separately

= se both objective stigmator selectors

± se the first one
± se the second one
± Refocus
± Repeat
 
 

 

= Resolving power is good (why?), but«
± To see image features contrast is needed

= ow to increase contrast«?

= (SEM contrast is derived from local topography

and/or differential interactions of beam with
sample)
 
 

 

= Mass-thickness Contrast (absorption)
± Density x thickness
± More dense or thicker areas look darker due
to absorption of beam electrons.
± Thickness fringes due to destructive
interference as beam traverses the sample
± se stains to highlight specific areas
= ranium, manganese, osmium
= Coat and/or shadow areas to generate contrast
 
 

 

= ther ³deficiency´ contrast mechanism

± Electron scattering
= Random
±
morphous materials change electron
trajectories

= Regular
± Crystalline materials change trajectories
uniformly
 
 

 

= Most of the beam is N S
± Goes right through the sample unperturbed

± ther elastic interactions change the direction

of N S electrons which can be selected for or
eliminated from the image forming beam.
More on this later«
 4 



 

= Type of source (W, La, E)
= igher mags means a strong nt-Proj lens,
which means lower intensity
= ard to see on fluorescent screen
= Ways to mitigate
± use lower mags
± converge beam with condenser lens
± align beam as needed
 4




 

= igher voltages produces
± shorter wavelength

± better resolution

± greater depth penetration of sample

 4




 

= Lower voltages produces
± greater contrast due to larger scattering angles
for slow(er) moving electrons

± less depth penetration

± larger proportion of electrons involved in

inelastic collision events
Electron eam-Sample nteraction
  
 


 

= igher mag-> features look larger (duh)

= ut intensity drops off and ability to

properly focus and stigmate do too

= se LWEST practical magnification

Most photographic emulsions used in electron microscopy
can resolve image details of ~×0µm, thus the resolution
of object details will depend on the image magnification
as shown in the table (resolution = ×0µm/magnification):

 
 

 
 


×,000 10.0
×0,000 1.0
50,000 0.4
100,000 0.×
  


 

= Low intensity situations lead to longer
exposure times

= Vibration will make edges blurry

= igh intensity situations lead to short

exposure times (and concomitant error)
 R 
R 

 


= igh intensity or long exposure situations

may cause sample to degrade (bonds break,
polymer chain-scission, etc.)
= Remember the contamination square in the
SEM??? Same thing happens in the TEM-
you will grow a nice carbon bump on
samples as you look at them.
cR 

  
 
 TEM Sample Prep for
iologicals
=
lmost always a microtome is involved
 
 


c

right ield Mode

Dark ield Mode

Diffraction Mode
 4 °
 

= f the main portion of the near-forward
scattered beam is used to form the image
± transmitted beam
± 000 beam
± zero-order beam
 Š  °
 

= f the transmitted beam is
excluded from the image
formation process
± off-axis imaging
± tilted beam imaging
c 

   
 

Š 

= Elastic Scattering Events

± ragg diffraction
= n¬=×d sin
 

Š 

= our conditions in ack ocal Plane ( P) of

the objective lens:

± No sample No reflections (only transmitted beam)

±
morphous Transmitted beam + random scattering
± Polycrystal Transmitted beam + rings
± Single crystal Transmitted beam + spots
 

Š 

ngle of incidence ~1/×0 to even come close to

satisfying the ragg condition.

Therefore only the lattice planes close to parallel to the

beam are involved in diffraction.
 

Š 

= Think of TEM as a
diffraction camera

Rd=¬L
R is measured Transmitted eam
d is the unknown L
¬ is the electron wavelength Diffracted eam
L is the camera length

¬L is the camera constant)

Reciprocal relationship between

lattice spacing and distance from R
the transmitted spot.
 

Š 

=
u (111) ring [×.35 Å d-spacing]

With ×00KV and L=5cm the (111)

ring should be at about 7.5mm from
the transmitted beam

Rd=¬L
R=0.0×7
50mm/×.35
   

c
 Electron Diffraction
TEM maging Modes: Diffraction vs 
Metal particles Polymer mix Electron Diffraction

TEM mages