Both
light microscopes
and electron microscopes use radiation (light or electron beams) to
form larger and more detailed images of objects (e.g. biological
specimens, materials, crystal structures, etc.) than the
human eye can produce unaided. (See also:
What is eyesight ?)
An electron microscope is a microscope that uses beams of
electrons instead of rays of
visible light
to form highly magnified images of tiny areas materials or biological
specimens. Comparing light vs electron microscopes is made more
complicated by the fact that there are different types of electron
microscopes.
The two main types of electron microscope are the Transmission Electron Microscope (TEM) and Scanning Electron Microscope
(SEM). Other types of electron microscope include the Scanning
Tunneling Microscope (STM) and Field Emission Transmission Microscope
(FE-TEM).
Brief notes, see the tables lower down this page for figures, explanations and comments:
Similarities: |
- Form larger (magnified) and more detailed (highly resolved) images of small objects or small areas of larger objects e.g. a leaf, part of a bone, etc. than can be formed by the human eye.
- Used in study and research in biology and medical sciences (more about histology), material sciences e.g. metallurgy and other aspects of science.
- Specimens must be carefully prepared using techniques appropriate for both the equipment and the sample e.g. slicing, staining, mounting, etc. (e.g. how to prepare histology slides).
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Differences: |
- Size: Light microscopes are smaller and lighter, so are easier to move and set-up.
- Cost / Availability: Light microscopes are less expensive than electron microscopes.
- Radiation Type: Light microscopes
use light (approx wavelength 400-700 nm), electron microscopes use beams
of electrons (approx equivalent wavelength 1 nm).
- Control of image formation : Light via glass lenses, beams of electrons can be focused using electromagnets due to negative charge on electrons.
- Resolution*: Electron microscopes have much higher resolution than light microscopes
- Magnification*: Electron microscopes have much higher magnification than light microscopes
- Colour Images: Light microscopes
form images including the range of wavelengths (colours) provided by the
light source - but remember that the colours seen are often due to
stains rather than the actual colours present in nature). Electron
microscopes produce greyscale (sometimes called "black and white")
images. However, "false-colour" electron micrographs are common - and
can be very beautiful!
- Preparation of specimens :
Generally involves harsher processes, e.g. using corrosive chemicals,
for viewing via electron microscope than preparation of slides for
viewing using a light microscope. Therefore more skill required - both
to prepare specimens and to interpret EM images (due to "artefacts" in
images, artefacts = features in images that
are not present in the specimen but rather are due to the processes used
to prepare the specimen and produce the image)
- Image Formation : Light microscope
images can be viewed directly. Electron microscopes require use of a
fluorescent screen, photographic plate or electronic display because
electrons cannot be observed directly by the human eye.
- Usage Limitations : Living
specimens cannot be viewed using electron microscopes because electron
microscopes require there to be a vacuum in the tube - otherwise the electrons would be absorbed by air molecules.
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* Differences due to the shorter equivalent wavelength of electron beams compared with visible light. |
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See the table at the bottom of this page for the advantages and disadvantages of light vs electron microscopes.
The following simple block diagram shows some of the
basic similarities between light microscopes and electron microscopes
(in general) by comparing the radiation pathways for a light microscope
with a general electron microscope.
Very simple block diagrams of Electron vs Light Microscopes
Compare Electron Microscopes vs Light Microscopes (Physical Aspects)
As there are different types of electron microscopes and capabilities
vary the following comparison of electron microscopes with light
microscopes is very general (including 'typical' rather than exact
figures).
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Light Microscope
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Electron Microscope
|
Radiation Type |
Visible light |
Beams of Electrons |
Approx. wavelength of radiation |
400–700 nm |
effective wavelength < 1 nm |
Radiation focussed by ... |
Lenses - usually glass lenses |
electromagnets |
Image formed by ... |
Light (of all colours whose wavelengths are supplied by the source of illumination) scatters from the various parts of the specimen
and some of that scattered light reaches the objective lens (above the
specimen) and is then re-directed through the objective and eyepiece
lenses to form a focussed image. |
Re. TEM: Selective absorption of electrons by the specimen.
(There must be a vacuum inside an operating electron microscope because
otherwise the electrons would be absorbed by air molecules before they
could reach the specimen.) Parts of the specimen absorb electrons and
therefore appear dark on the micrograph, while other areas of the
specimen allow electrons to pass through - causing those areas to appear
bright on the micrograph. |
Image formed on ... |
The human
eye can view a real image directly by looking into the eyepiece of the
microscope. An image is then projected onto the retina of the eye. |
An image is
formed by projecting a focussed image of the specimen onto a surface
coated with electron-sensitive compounds. It may be labelled
"fluorescent screen" on simple diagrams. |
Typical resolution |
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Reported
values vary from a typical 0.2-0.5 nm for TEMs, through 0.1 nm for STM,
down to 50 pm (= 0.5 ångström = 0.05 nm) for FE-TEM.
1 pm = 1 picometre is 10-12m
1 Å (1 ångström) = 1 x 10-10m
|
200 nm |
0.5 nm is the figure cited in AS Biology books |
1 nanometre is 10-9m so
200 nm = 200 x 10-9m = 2 x 10-7m |
1 nm (1 nanometre)
= 1 x 10-9m, so
0.5 nm
= 0.5 x 10-9m = 5 x 10-8m |
Compare the resolution of an electron microscope with that of a light microscope: |
resolution of electron microscope
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=
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0.5 nm
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=
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1
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...
means that in very general terms, an electron microscope can detect and
display around 400x as much detail as a light microscope, e.g. by
imaging a line of 200 particles spaced with gaps whose size is equal to
the particle size when the particle size is so small that the entire
line is seen a single item when using a light microscope.
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resolution of light microscope
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200 nm |
400 |
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Typical useful magnification |
Sources state between x1000 and x2000
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Some sources state up to x10,000,000
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less than x2000 usable image quality
"at best" x1500 useful image quality
typically up to x1000 |
approx x100,000 in SEM
approx x250,000 in TEM |
Compare the magnification of an electron microscope with that of a light microscope: |
magnification of electron microscope
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= |
100,000x
|
|
...
means that in very general terms, an electron microscope can produce
magnifications of at least 100 times greater than a typical light
microscope
|
magnification of light microscope
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1000x
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Note: The figures used in the equations are indicated in red above.
Notice the other values that could be used for this type of very general comparison.
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Above: Table comparing the physics of light vs electron microscopes
Compare the Advantages and Disadvantages (i.e. limitations of use) of Electron Microscopes vs Light Microscopes
Due to constraints imposed by the ways in which different types of
microscopes produce images, some microscopes can be used in certain ways
that others cannot. For example, it is not possible to observe
individual molecules using a light microscope or to watch living
processes happen using an electron microscope.
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Light Microscope (LM)
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Electron Microscope (EM)
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1. |
Living cells & tissues |
Can watch living processes take place e.g. microscopic pond life in action, and even cell division. |
Not possible to view any living material due to vacuum inside EM |
2. |
Thickness of specimen |
Specimen must be thin but can adjust focus to different positions (heights) within thin specimen on glass slide |
Very thin sections only in TEM
Images surfaces (only) in SEM |
3. |
Depth: 2D or 3D ? |
Image plane approx "flat" (2D) but, as above, can adjust focus through specimen |
2D only in TEM ;
SEM images surfaces - hence shows depth info that seems like 3D |
4. |
Specimen preparation
/ artefacts |
Simpler preparation (staining still required) |
Harsher preparation procedures incl. use of corrosive chemicals that may cause "artefacts" in the resulting micrographs |
5. |
Magnification |
Lower Magnification |
Higher Magnification - so several micrographs may be needed per specimen |
6. |
Resolution |
Lower Resolution |
Higher Resolution - good for measuring sizes of smaller features |
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Above: Table comparing use of light vs electron microscopes
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