Abdul Rajeeb L.
Lucman 05/09/17
Grade 7-Emerald Sir Don
Lariosa
MICROSCOPY
Microscopy is the technical field of using microscopes to view objects
and areas of objects that cannot be seen with the naked eye. There are three
well-known branches of microscopy: optical, electron, and scanning probe
microscopy.
Optical & electron microscopy involve the diffraction, reflection, or
refraction of electromagnetic radiation interacting with the specimen, and
the collection of another signal in order to create an image. This process may
be carried out by wide-field irradiation of the sample, or by scanning of a fine
beam over the sample. Scanning probe microscopy involves the interaction
of a scanning probe with the surface of the object.
The development of microscopy revolutionized biology, gave rise to the
field of histology and so remains an essential technique in the life and
physical sciences.
Optical Microscopy
Optical or light microscopy involves passing visible light reflected from
the sample through a single lens or multiple lenses to allow a magnified view
of the sample. The resulting image can be detected directly by the eye,
imaged on a photographic plate or captured digitally. The single lens, or the
system of lenses with its imaging equipment, along with the appropriate
lighting equipment, sample stage and support, makes up the basic light
microscope.
Figure 1: An optical microscope. Figure 2: Light
reflection between two optical microscopes.
Electron Microscopy
Electron microscopy is a technique for obtaining high resolution images
of biological and non-biological specimens. It is used in biomedical research
to investigate the detailed structure of tissues and cells. The high resolution
of EM images results from the use of electrons (which have very short
wavelengths) as the source of illuminating radiation. Electron microscopy is
used in conjunction with a variety of ancillary techniques to answer specific
questions. EM images provide key information on the structural basis of cell
function and of cell disease.
There are two main types of electron microscope the transmission EM
and the scanning EM. The transmission electron microscope is used to view
thin specimens (tissue sections, molecules) through which electrons can pass
through, generating a projection image. The TEM is analogous in many ways
to the compound light microscope. TEM is used, among other things, to
image the interior of cells, and the organization of molecules in viruses and
cytoskeletal filaments.
Figure 3: A transmission electron microscope.
Conventional scanning electron microscopy depends on the emission
of secondary electrons from the surface of a specimen. Because of its great
depth of focus, a scanning electron microscope is the EM analog of a stereo
light microscope. It provides detailed images of the surfaces of cells and
whole organisms that are not possible by TEM. It can also be used for particle
counting and size determination, and for process control. It is termed a
scanning electron microscope because the image is formed by scanning a
focused electron beam onto the surface of the specimen in a raster pattern.
The interaction of the primary electron beam with the atoms near the
surface causes the emission of particles at each point in the raster. These
can be collected with a variety of detectors, and their relative number
translated to brightness at each equivalent point on a cathode ray tube.
Because the size of the raster at the specimen is much smaller than the
viewing screen of the CRT, the final picture is a magnified image of the
specimen. Appropriately equipped SEMs can be used to study the
topography and atomic composition of specimens.
Figure 4: A scanning electron microscope. Figure 5: Pollen grain under
SEM (left) and TEM (right).
Scanning Probe Microscopy
Scanning probe microscopy covers several related technologies for
imaging and measuring surfaces on a fine scale, down to the level of
molecules and groups of atoms. At the other end of the scale, a scan may
cover a distance of over 100 micrometers in the x and y directions and 4
micrometers in the z direction. This is an enormous range. It can truly be
said that the development of this technology is a major achievement, for it is
having profound effects on many areas of science and engineering. SPM
technologies share the concept of scanning an extremely sharp tip across the
object surface. The tip is mounted on a flexible cantilever, allowing the tip to
follow the surface profile. When the tip moves in proximity to the
investigated object, interaction between the tip and the surface affect the
movement of the cantilever. These movements are detected by selective
sensors. Various interactions can then be studied depending on the
mechanics of the probe.
Figure 6 and 7: A scanning probe microscope (left) and the tip, or probe used to
interact with the specimen.