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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.