Light Microscope (Types)

Light microscope: Is the type of microscope that uses visible light and a system of lenses to magnify
images of small subjects. Some of major types of light microscope are Dark field microscope (ultra-
microscope), Phase-contrast microscope, Fluorescent microscope, confocal microscope and Polarized
microscope. These types of light microscope are explained as follow:
A. DARK FIELD MICROSCOPY.
Is the one that is used to examine live microorganisms that are either invisible in ordinary light
microscope, cannot be stained by standard methods or are so distorted by staining that their
characteristic then cannot be identified.
Principle:
The dark ground microscope creates a contrast between the object and the surrounding field, such that
background is dark and the background is bright. The objective and the ocular lenses used the same as
in ordinary light microscope, however special condenser is used which prevents the transmitted light
from directly illuminating the specimen.
Applications:
 Used for diagnosis of Syphilis.

 Used in viewing blood cells.
 Used in viewing different types of bacteria.
 Used in viewing hairline metal fracture.
 Used in viewing diamonds and other precious stones.
 In viewing bacteria.
B. PHASE CONTRAST MICROSCOPE
Is the one that made it possible to study living cells and how they proliferate through cell division. It is
used to examine living organisms or specimens that that would be damaged or altered by attaching
them to slides or staining. It uses conventional light microscope fitted with phase-contrast objective and
phase-contrast condenser.
Principle:
Unstained bacteria have constituent of different refractive index.
Diffraction of light.
Phase contrast microscope employs an optical mechanism to translate minute variations in phase into
corresponding changes in intensity of image.
Applications:
 Used to provide contrast of transparent specimens such as living cells or small organisms.
 Unstained paraffin, resin and frozen sections.
C. FLOURESENT MICROSCOPE:
Is the type of light microscope that uses higher intensity light to illuminate the sample. This light excites
fluorescence species in the sample, which then emit light of longer wavelength.
Principle:
The specimen is illuminated with light of specific wavelength. Which is absorbed by the fluorophores,
causing them to emit light of longer wavelength i.e. of a principle different color than absorbed light.
The illumination light is separated from the much weaker emitted fluorescence through the use spectral
emission filter.
Applications:
 Fluorescence microscopy can be applied to detect particles below the resolution of light
microscope.
 Fluorescent microscopy is the basis for immunofluorescence technique for demonstration of
antigens and antibodies in tissue and sera.
D. CONFOCAL MICROSCOPE:
Is the optical imaging technique for increasing optical resolution and contrast of a micrograph by means
of adding a spatial pinhole placed at a confocal plane of a lens to eliminate out of focus light.
Principle:
A pinhole placed in front of the illumination source to allow transmission only through a small area.
The illumination pinhole is imaged onto the focal plane of the specimen i.e. only a point of a specimen is
illuminated at one time.
Fluorescence excited at this manner is imaged onto a confocal pinhole placed in front of the detector.
Only fluorescence excited within a focal plane of the specimen will go through the detector pinhole.
Need to scan point onto the sample.
Applications:
 Stem cell research.

 Photobleaching studies.
 Fluorescence resonance energy transfer (FRET)
 Multiphoton microscopy.
 DNA hybridization.
E. POLARIZED LIGHT MICROSCOPE:
Is the one that is designed to observe optically anisotropic specimen (BIREFRINGENCE SPECIMEN).
Equipped with polarizer and analyzer. It works on principle of interference.
Principle:
A polarizer only allows certain light waves or vibrations to pass through it. An analyzer often second
polarizer located above the sample determines amount and direction of light that illuminates the
sample.
Light that is confirmed to a single plane of vibration is said to be plane polarized.
The examination of the interaction of plane polarized light with matter is made possible with the
polarizing microscope.
Applications:
 Used in the field of geology or petrography for the study of rocks and minerals.
 Used in Histopathology e.g. Muscle striations, charcot-layden crystals(Miscellaneous)
 Used in determination of refractive index that help to identify minerals present in a soil
sample or the identity of man made fiber.

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Light microscope: Is the type of microscope that uses visible light and a system of lenses to magnify

A. DARK FIELD MICROSCOPY.

 Used for diagnosis of Syphilis.

B. PHASE CONTRAST MICROSCOPE

Unstained bacteria have constituent of different refractive index.

 Stem cell research.

E. POLARIZED LIGHT MICROSCOPE:

Light that is confirmed to a single plane of vibration is said to be plane polarized.

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