In 1932 Knoll and Ruska invented first electron microscope.
The electron microscope uses a beam of electrons rather than visible light.
The magnified image is visible on a fluorescent screen and can be recorded on a
photographic film.
The drawback of the electron microscope is specimen are killed in order to view the cells or
organisms.
Images produced by electrons lack color, electron micrographs are always shades of black,
gray, and white.
Two general forms of EM are the transmission electron microscope (TEM) and the
scanning electron microscope (SEM).
Transmission electron microscopes are the method of choice for viewing the detailed
structure of cells and viruses.
This microscope produces its image by transmitting electrons through the
specimen.
Because electrons cannot readily penetrate thick preparations, the specimen must
be sectioned into extremely thin slices (20–100 nm thick) and stained or coated with metals
that will increase image contrast.
The darkest areas of TEM micrographs represent the thicker (denser) parts, and the
lighter areas indicate the more transparent and less dense parts.
Transmission electron microscope (TEM)-Transmission electron microscope is complex and
sophisticated. In TEM a heated tungsten filament in the electron gun generates a beam of
electrons that is focused on the specimen the condenser, since electrons cannot pass
through a glass lens, doughnut shaped electromagnets called magnetic lenses are used to
focus the beam. The column containing the lenses and specimen must be under high
vacuum to obtain a clear image because electrons are reflected by collisions with air
molecules. The specimens scatters electrons passing through it and the beam is focused by
magnetic lenses to form an enlarged, visible image of thespecimen on the fluorescent
screen. The screen can be moved a side and the image captured on photographic film as a
permanent record.
2) Scanning electron microscope(SEM)- Scanning electron microscope has been used to
examine the surfaces of microorganisms.
Microorganisms must first fixed, dehydrated, and dried to preserve surface structure and
prevent collapse of the cells. when they are exposed to SEM's high vacuum.
The SEM scans a narrow tempered electron beam back and forth over the specimen.
When the beam strikes a particular area surface atoms discharge a tiny shower of electrons
called secondary electrons and these are trapped by a special detector.
Secondary electrons entering the detector strike a scintillator causing it to emit light flashes
that a photo multiplier converts to an electrical current and amplifiers.
The signal is sent to a cathode ray tube and produces an image like a television picture
which can be viewed or photographed.
A realistic 3-dimensional image of the microorganism's surface with greater depth of focus
[Link] specimen is placed in the vacuum chamber and covered with a thin coat of gold.
The electron beam then scans across the specimen and knocks loose showers of electrons
that are captured by a detector. An image builds line by line, as in a television receiver.
Electrons that strike a sloping surface yield fewer electrons, thereby producing a darker
contrasting spot and a sense of three dimensions.
The resolving power of the conventional SEM is about 10 nm and magnifications with the
SEM are limited to about 20,000x.