Image modes

The Origin of Contrast

 The electron beam interacts with a thin-film specimen both elastically and inelastically -
elastic interactions that dominate the contrast
-inelastic scattering events that contain information on the chemical composition of the
sample
 Contrast arises from three quite distinct image-forming processes, and these are termed
-mass–thickness contrast
-diffraction contrast and
-phase contrast
 Mass–thickness contrast
 The probability of an electron being elastically scattered out of the incident beam
depends on the atomic scattering factor, which increases monotonically with the atomic
number and the total number of atoms in the path of the beam, that is the total thickness
of the film. Mass–thickness contrast thus reflects a combination of variations in
specimen thickness and specimen density.

 The simplest mode of image formation occurs when some areas of the sample absorb
more electrons than other parts.

 The intensity of the transmitted beam

(It) is the intensity of primary beam (Io)
less the intensity of beam deflected by
object (Id) in a specimen.

 The contrast (C) in a TEM is defined by

Applications:

 In the life sciences, mass–thickness contrast almost always dominates the image. The
contrast of soft tissue, biological samples in the electron microscope is often enhanced by a
heavy-metal, tissue-staining procedure.

 In the natural sciences and in engineering studies, mass–thickness contrast only

predominates in non-crystalline materials, such as two-phase glasses, as well as in replica
studies, where the contrast is often due to variations in the thickness of a metal shadow
deposited on the replica, or to the presence of an extracted
phase.
 Besides staining, common effective techniques to
increase contrast include using a smaller objective
aperture and using a lower acceleration voltage.
The negative side of using such techniques is
reduction of total brightness.
Diffraction Contrast
Diffraction contrast is the primary mechanism of TEM image formation in crystalline specimens.
Diffraction can be regarded as collective deflection of electrons. Electrons can be scattered
collaboratively by parallel crystal planes similar to X-rays.

The intensity of the transmitted beam is reduced when the objective aperture blocks
the diffraction beams, similar to the situation of mass–density contrast.

The main difference between the two contrasts is that the diffraction contrast is very
sensitive to specimen tilting in the specimen holder but mass–density contrast is
only sensitive to total mass in thickness per surface area.
The diffraction angle (2θ) in a TEM is very small (≤10) and the diffracted beam from a
crystallographic plane (hkl) can be focused as a single spot on the back-focal plane of the
objective lens. The Ewald sphere is particularly useful for interpreting electron
diffraction in the TEM. When the transmitted beam is parallel to a crystallographic axis,
all the diffraction points from the same crystal zone will form a diffraction pattern (a
reciprocal lattice) on the back-focal plane.
 The diffraction contrast can generate bright-field and dark-field TEM images.

 A TEM can be operated in two modes:

o the image mode and
o the diffraction mode
 The modes can be changed by changing the optical path in the TEM, as the following Figure.
  In the image mode, the image of the specimen is focused and projected onto a fluorescent
screen.

 In the diffraction mode, the diffraction

pattern formed on the back-focal plane is
projected onto a fluorescent screen.

Optical arrangement of bright- and dark-field contrast

 A bright-field image is obtained by allowing only the transmitted beam to pass

the objective aperture, exactly the same as in mass–density contrast
 A dark-field image is obtained by allowing one diffraction beam to pass the objective
aperture
 To switch between bright- and dark-field modes, we need to operate in the diffraction
mode first in order to select one diffraction spot in the diffraction pattern by moving the
aperture to the center of that spot.

 In the TEM, the direction of the incident beam is moved, rather than the aperture, to shift the
selected diffraction spot to the central axis.

Diffraction contrast is very

sensitive to specimen
tilting. Even a tilt of less
than1◦ can cause the
contrast to change from
bright to dark.
Dark-field is not as commonly used as bright-field imaging. However, the dark field image
may reveal more fine features that are bright when the background becomes dark.

Interestingly, there are several dark-field

images corresponding to a single bright-
field image because we can select different
diffraction spots (hkl) to obtain a dark-field
image.
Phase contrast
 TEM can also use the phase difference in electron waves to generate contrast. The phase-
contrast mechanism, however, is much more complicated than that of light microscopy. The
TEM phase contrast produces the highest resolution of lattice and structure images for
crystalline materials. Thus, phase contrast is often referred as to high-resolution
transmission electron microscopy (HRTEM).

 Phase contrast must involve at least two electron waves that are different in wave
phase.

 At least two beams are needed (the transmitted beam and a diffraction beam) to
participate in image formation in a TEM.

Both mass–density and diffraction contrasts

are amplitude contrast
Selected Area Diffraction
 Electron diffraction is not only useful to generate images of diffraction contrast, but also
for crystal structure analysis, similar to XRD methods.

Selected-Area Diffraction Characteristics:

 The diffraction angle in the TEM
is very small (commonly θ < 1◦), because
the reflecting lattice planes are nearly
parallel to the primary beam.

 Using the approximation sin θ ≈ θ for a small angle,

we can rewrite Bragg’s Law as the following.
where L is the camera length of a TEM, the distance between the crystal and
photographic plate of camera. In fact, L is an equivalent camera length for calculating
the spacing of the lattice plane. λL is called as the camera constant of a TEM
Single-crystal diffraction

 To obtain a diffraction pattern, we need to use the selected-area aperture to isolate a single
crystal in the image mode, and then switch to the diffraction mode.

 Tilting the specimen is necessary to obtain a diffraction pattern with high symmetry with
respect to its central transmitted spot.
 After obtaining a diffraction pattern, we may want to index the pattern in order
to determine the crystal structure or crystal orientation that the pattern represents.

 Indexing a pattern means to assign a set of h, k, and l to each diffraction spot in

the pattern.
 The information can be directly extracted from a pattern that includes the length of R for
each spot and angles between any two R vectors.
Indexing a Cubic Crystal Pattern

In the pattern, the central spot of the transmitted beam is brightest and should be
indexed as (0 0 0) of the reciprocal lattice plane.

–select two spots: m and n and the R lengths of m and n are measured as 8.92 and
12.6 mm, respectively; and the angle between Rm and Rn is measured as 45◦.
λ for 200 kV is 0.00251 nm; the camera length (L) of 1.0
m; the lattice parameter of NaCl (a) is 0.563 nm;
=
-determine that the Rm matches and Rn matches
Then, check whether the angle (RmRn) matches
that between specific planes.