Types of Microscopes

What are the Different Types of Microscopes and Their Uses?

There are a number of different types of microscopes and each of them solves
unique problems. Below you will find information on the five different types of
microscopes along with the applications for each microscope and just who
might use each instrument. Below each description of the microscope and its
use is an image that was captured using that particular microscope.

5 Different Types of Microscopes:

1. Stereo Microscope
2. Compound Microscope
3. Inverted Microscope
4. Metallurgical Microscope
5. Polarizing Microscope
 
Stereo Microscopes

Stereo microscopes are used to look at a variety of samples that you would be

able to hold in your hand. A stereo microscope provides a 3D image or "stereo"
image and typically will provide magnification between 10x - 40x. The stereo
microscope is used in manufacturing, quality control, coin collecting, science,
for high school dissection projects, and botany. A stereo microscope typically
provides both transmitted and reflected illumination and can be used to view a
sample that will not allow light to pass through it.

The following are samples often viewed under a stereo microscope: coins,
flowers, insects, plastic or metal parts, printed circuit boards, fabric weaves,
frog anatomy, and wires.

This image of a penny was captured under the a coin collecting stereo zoom
microscope at 20x magnification.
Compound Microscopes

A compound microscope may also be referred to as

a biological microscope. Compound microscopes are used in laboratories,
schools, wastewater treatment plants, veterinary offices, and for histology and
pathology. The samples viewed under a compound microscope must be
prepared on a microscope slide using a cover slip to flatten the sample. Students
will often view prepared slides under the microscope to save time by
eliminating the slide preparation process.

The compound microscope can be used to view a variety of samples, some of

which include: blood cells, cheek cells, parasites, bacteria, algae, tissue, and
thin sections of organs. Compound microscopes are used to view samples that
can not be seen with the naked eye. The magnification of a compound
microscope is most commonly 40x, 100x, 400x, and sometimes 1000x.
Microscopes that advertise magnification above 1000x should not be purchased
as they are offering empty magnification with low resolution.

This image of mushroom spores was captured under a compound biological

microscope at 400x magnification.
Inverted Microscopes

Inverted microscopes are available as biological inverted

microscopes or metallurgical inverted microscopes. Biological inverted
microscopes provide magnification of 40x, 100x and sometimes 200x and 400x.
These biological inverted microscopes are used to view living samples that are
in a petri dish. An inverted microscope allows the user to place the petri dish on
a flat stage, with the objective lenses housed beneath the stage. Inverted
microscopes are used for in-vitro fertilization, live cell imaging, developmental
biology, cell biology, neuroscience, and microbiology. Inverted microscopes are
often used in research to analyze and study tissues and cells, and in particular
living cells.

Metallurgical inverted microscopes are used to examine large parts at high

magnification for fractures or faults. They are similar to biological inverted
microscope in the magnification provided, but one primary difference is that the
samples are not placed in a petri dish, but rather a smooth side of the sample
must be prepared so it can lay flat on the stage. This smooth sample is polished
and is sometimes referred to as a puck.
Metallurgical Microscopes

Metallurgical microscopes are high power microscopes designed to view

samples that do not allow light to pass through them. Reflected light shines
down through the objective lenses providing magnification of 50x, 100x, 200x,
and sometimes 500x. Metallurgical microscopes are utilized to examine micron
level cracks in metals, very thin layers of coatings such as paint, and grain
sizing.

Metallurgical microscopes are utilized in the aerospace industry, the automobile

manufacturing industry, and by companies analyzing metallic structures,
composites, glass, wood, ceramics, polymers, and liquid crystals.

This image of a piece of metal with scratches on it was captured under a

metallurgical microscope at 100x magnification.
Polarizing Microscopes

Polarizing microscopes use polarized light along with transmitted and, or

reflected illumination to examine chemicals, rocks, and minerals. Polarizing
microscopes are utilized by geologists, petrologists, chemists, and the
pharmaceutical industry on a daily basis.

All polarizing microscopes have both a polarizer and an analyzer. The polarizer
will only allow certain light waves to pass through it. The analyzer determines
the amount of light and direction of light that will illuminate the sample. The
polarizer basically focuses different wavelengths of light onto a single plane.
This function makes the microscope perfect for viewing birefringent materials.
This is Vitamin C captured under a polarizing microscope at 200x
magnification.
Light Microscope- definition, principle,
types, parts, magnification
Table of Contents  
History of Microscopy: Overview

What is a light microscope?

Figure: Diagram of Light Microscopes, created with biorender.com

Principle of a light microscope (optical microscope)

Types of light microscopes (optical microscope)

Brightfield Light Microscope (Compound light microscope)

Parts of a bright-field microscope (Compound light microscope)

Magnification by Bright field Microscope (Compound light microscope)

Applications of the Bright Field Light Microscope (Compound light microscope)

Light Microscope Free Worksheet

Phase Contrast Microscope

Parts of the Phase Contrast Microscope

The functioning of the Phase Contrast microscope

Applications of Phase-Contrast Microscope

Dark-Field Light Microscope

Applications of the Dark Field Microscope

The Fluorescent Microscope

Applications of the Fluorescent Microscope

References

Light Microscope (Optical microscope)

History of Microscopy: Overview
 The evolution of the Microbiology field put to perspective the need to
identify, view, observe and understand microorganisms, including
their structural morphologies and mechanisms. Microbiology’s scope
is to study organisms and minute agents that can only be examined
and observed with a microscope.
 Although scientifically, the first simple microscope was discovered by
two Dutch scientists, Zaccharias Janssen and his father, Hans who
made spectacles, were the first to experiment with their lenses by
combining lenses in a tube and observed that the objects that were
nearby, appeared closer and larger. Despite not being included as a
scientific discovery, this act paved the way for scientific evolution.
 From the History of Microbiology, Antony Van Lewnehoueek an
amateur Microbiologist made the first simple microscope, that
enabled him to observe the presence of tiny living organisms in pond
water that appeared like dots. His simple microscope was made up of
a double convex glass lens that was held between two silver plates.
 The application of microscopy in Microbiology enhanced the
visualization of cells and microorganisms by magnifying their images
to make them larger.
The light microscope is also known as an optical
microscope.
What is a light microscope?
 A light microscope is a biology laboratory instrument or tool, that
uses visible light to detect and magnify very small objects, and
enlarging them.
 They use lenses to focus light on the specimen, magnifying it thus
producing an image. The specimen is normally placed close to the
microscopic lens.
 Microscopic magnification varies greatly depending on the types and
number of lenses that make up the microscope. Depending on the
number of lenses, there are two types of microscopes i. e Simple light
microscope (it has low magnification because it uses a single lens)
and the Compound light microscope (it has a higher magnification
compared to the simple microscope because it uses at least two sets
of lenses, an objective lens, and an eyepiece). The lenses are aligned
in that, they can be able to bend light for efficient magnification of
the image.
 The functioning of the light microscope is based on its ability to focus
a beam of light through a specimen, which is very small and
transparent, to produce an image. The image is then passed through
one or two lenses for magnification for viewing. The transparency of
the specimen allows easy and quick penetration of light. Specimens
can vary from bacterial to cells and other microbial particles.
Figure: Diagram of Light Microscopes.

Principle of a light microscope (optical

microscope)
As mentioned earlier, light microscopes visualize an image by using a
glass lens and magnification is determined by, the lens’s ability to bend
light and focus it on the specimen, which forms an image. When a ray of
light passes through one medium into another, the ray bends at the
interface causing refraction. The bending of light is determined by
the refractive index, which is a measure of how great a substance slows
the speed of light. The direction and magnitude of the bending of the light
are determined by the refractive indexes of the two mediums that form
the interface.
A medium with a lower refractive index such as glass to air, it normally
speeds up the light penetration and making light bend away from the
normal and when light is passed through a medium with a greater
refractive index such as air to glass, it normally slows down and bends
towards the normal, perpendicularly to the surface.
If an object is put between these two mediums i.e between water and air,
in this case, a prism, the prism will bend the light at an angle. This is how
the microscopic lenses work, they bend the light at an angle. The lens
(convex) on receiving the light rays, it focuses the rays at a specific point
known as the focal point (F-point). The measure of distance from the
center of the lens and the focal point is known as the focal length.
A microscope uses lenses whose strength is predetermined, in that, the
strength of a lens is directly related to the focal length i.e short focal
length magnifies objects more than lenses with a long focal length.
Microscopy works strictly with a factor of resolution whereby resolution
being the ability of a lens to be able to differentiate small objects that are
closely packed together. The resolution of a light microscope is
determined by a numerical aperture of its lens system and by the
wavelength of the light it employs; a numerical aperture a definition of
the light wavelengths produced when the specimen is illuminated.
A minimum distance (d) between two objects that distinguishes then to
be two separate entities, determined by the wavelengths of the light can
be calculated by an Abbe equation using the wavelength of the light that
illuminated the specimen (Lambda, λ) and the numerical aperture (NA, n
sin Ɵ) i.e. d=0.5 λ/n sin Ɵ

Types of light microscopes (optical

microscope)
With the evolved field of Microbiology, the microscopes
used to view specimens are both simple and compound light microscopes,
all using lenses. The difference is simple light microscopes use a single
lens for magnification while compound lenses use two or more lenses for
magnifications. This means,  that a series of lenses are placed in an order
such that, one lens magnifies the image further than the initial lens.
The modern types of Light Microscopes include:
1. Bright field Light Microscope
2. Phase Contrast Light Microscope
3. Dark-Field Light Microscope
4. Fluorescence Light Microscope

Brightfield Light Microscope (Compound

light microscope)
 This is the most basic optical Microscope used in microbiology
laboratories which produces a dark image against a bright
background. Made up of two lenses, it is widely used to view plant
and animal cell organelles including some parasites such
as Paramecium after staining with basic stains.
 Its functionality is based on being able to provide a high-resolution
image, which highly depends on the proper use of the microscope.
This means that an adequate amount of light will enable sufficient
focusing of the image, to produce a quality image.
 It is also known as a compound light microscope.
Parts of a bright-field microscope (Compound light
microscope)

Figure created with biorender.com

It is composed of:
 Two lenses which include the objective lens and the eyepiece or
ocular lens.
 Objective lens is made up of six or  more glasses, which make the
image clear from the object
 The condenser is mounted below the stage which focuses a beam of
light onto the specimen. It can be fixed or movable, to adjust the
quality of light, but this entirely depends on the microscope.
 They are held together by a sturdy metallic curved back used as
an arm and a stand at the bottom, known as the base, of the
microscope. The arm and the base hold all the parts of the
microscope.
 The stage where the specimen is placed, allowing movement of the
specimen around for better viewing with the flexible knobs and it is
where the light is focused on.
 Two focusing knobs i.e the fine adjustment knob and the coarse
adjustment knob, found on the microscopes’ arm, which can move
 the stage or the nosepiece to focus on the image.  the sharpen the
image clarity.
 It has a light illuminator or a mirror found at the base or on the
microbes of the nosepiece.
 The nosepiece has about three to five objective lenses with different
magnifying power. It can move round to any position depending on
the objective lens to focus on the image.
 An aperture diaphragm also is known as the contrast, which controls
the diameter of the beam of light that passes through the condenser,
in that, when the condenser is almost closed, the light comes through
to the center of the condenser creating high contrast. But when the
condenser is widely open, the image is very bright with very low
contrast.
Magnification by Bright field Microscope (Compound light
microscope)
During visualization, the objective lens remains parfocal which means,
when the objective lens is changed, the image still remains in focus.  The
objective lens plays a major role in focusing the image on the condenser
forming an enlarged clear image within the microscope, which is then
further magnified by the eyepiece to a primary image.
What is seen in the microscope as an enlarged clear image of the
specimen is known as the virtual image. To calculate the magnification,
multiply the objective and eyepiece objective magnification together. The
magnification is standard, i.e not too high nor too low, and therefore
depending on the magnification power of the lenses, it will range between
40X and 100oX.
Calculation of magnification = Magnification of objective
lens/magnification of the eyepiece lens
The objective lens plays a vital role in not only enlarging the image but
also making it clear for viewing, a feature known
as resolution. Resolution according to Prescott, is the ability of a lens to
separate or distinguish between small objects closely linked together.
Whereas the eyepiece magnifies the image at the end of the viewing, its
magnification range is lower than that of the objective lens at 8X-12X
(10X standard) and that of the objective lens at 40X-100X, magnification,
and resolution of the microscope is highly dependant on the objective
lens.
Applications of the Bright Field Light Microscope
(Compound light microscope)
Vastly used in Microbiology,  this microscope is used to view fixed and live
specimens, that have been stained with basic stains. This gives contrast
for easy visibility under the microscope. Therefore it can be used to
identify basic bacteria cells and parasitic protozoans such as Paramecium.
Light Microscope Free Worksheet
Phase Contrast Microscope
 This is a type of optical microscope whereby small light deviations
know as phase shifts occur during light penetration into the
unstained specimen. These phase shifts are converted into the image
to mean, when light passes through the opaque specimen, the phase
shifts brighten the specimen forming an illuminated (bright) image in
the background.
 The phase-contrast microscope produces high contrast images when
using a transparent specimen more so those of microbial cultures,
thin tissue fragments, cell tissues, and subcellular particles.
 The principle behind the working of the phase-contrast microscope is
the use of an optical method to transform a specimen into an
amplitude image, that’s viewed by the eyepiece of the microscope.
 The PCM can be used to view unstained cells also known as
the phase objects, which means that the morphology of the cell is
maintained and the cells can be observed in their natural state, in
high contrast and efficient clarity. This is because if the specimens are
 stained and fixed, they kill most cells, a characteristic that is uniquely
undone by the brightfield light microscope.
 The shifts that occur during light penetration, become converted to
changes in amplitude which causes the image contrast.
 Coupled with contrast-enhancing elements such as fluorescence, they
produce better visuals of the specimens’ image.
Parts of the Phase Contrast Microscope
The instrumentation of the Phase Contrast Microscope is based on its light
pathways from receiving the source of light to the visualization of the
image.
Therefore its sequentially made up of:
 Light source (Mercury arc lamp)
 Collective lens
 Aperture
 Condenser
 Condenser annular
 Specimen
 Objective
 Phase plate
 Deflected light
 Phase ring
The functioning of the Phase Contrast microscope
 The change caused by the deviated scattered (Deflected) light and
the undeviated light that reaches the specimen which is absorbed,
create at a certain wavelength, producing color. The difference
created by the scattered light and that of the absorbed light is known
as amplitude variations. These amplitude variations are sensitive to
allowing visualization by photographic equipment like the Phase
Contrast Microscope, hence seen by the human eye.
 The Condenser of the phase-contrast microscope has an opaque disk
that is known as an annular ring, with a transparent ring that
produces a cone of light, that passes through a specimen. Due to light
variations some light bend at the specimen, caused by variations in
light density, forming an image at the objective lens. The undeviated
light will strike the phase ring on the phase plate and the deviated
light will miss the phase ring passing through the phase plate directly,
this forms an image.
The Phase-Contrast Microscope is designed with objective lenses that
have the ability to perform multiple functions when combined with
contrast-enhancing techniques, for example, fluorescence. The objective
lenses are located in the internal phase plate with variation in the light
absorption and phase displacement i.e undiffraction, creating a wide
spectrum for contrasting the specimen and forming a strong contrast in
the background.
Applications of Phase-Contrast Microscope
 Determine morphologies of living cells such as plant and animal cells
 Studying microbial motility and structures of locomotion
 To detect certain microbial elements such as the bacterial
endospores.
Dark-Field Light Microscope
This is a specialized type of bright field light microscope which has several
similarities to the Phase-Contrast  Microscope. To make a dark field
Microscope, place a darkfield stop underneath and a condenser lens which
produces a hollow cone beam of light that enters the objective only, from
the specimen (Prescott, pg 22).
This technique is used to visualize living unstained cells. This is effected
by the way illumination is done on the specimen in that, when a hollow
cone beam of light is transmitted to the specimen, deviated light
(unreflected/unrefracted) rays do not pass through the objectives but the
undeviated (reflected/refracted) light passes through the objectives to the
specimen forming an image.
This makes the surrounding field of the specimen appear black while the
specimen will appear illuminated. This is enabled by the dark background
this the name, dark-field Microscopy.
Applications of the Dark Field Microscope
 It is used to visualize the internal organs of larger cells such as the
eukaryotic cells
 Identification of bacterial cells with distinctive shapes such
as Treponema pallidum, a causative agent of syphilis.

The Fluorescent Microscope

The above-discussed microscopes will normally produce images after a
light has been transmitted and passed through the specimen.
In the case of the fluorescent Microscope, the specimen emits light. How?
By adding a dye molecule to the specimen. This dye molecule will
normally become excited when it absorbs light energy, hence it releases
any trapped energy as light. The light energy that is released by the
excited molecule has a long wavelength compared to its radiating light.
The dye molecule is normally a fluorochrome, that fluoresces when
exposed to the light of a certain specific wavelength. The image formed is
a fluorochrome-labeled image from the emitted light
The principle behind this working mechanism is that the fluorescent
microscope will expose the specimen to ultra or violet or blue light, which
forms an image of the specimen that is emanated by the fluorescent light.
They have a mercury vapor arc lamp that produces an intense beam of
light that passes through an exciter filter. The exciter filter functions to
transmit a specific wavelength to the fluorochrome stained specimen,
producing the fluorochrome-labeled image, at the objective.
After the objective, there is a barrier filter that functions primarily to
remove any ultraviolet radiation that may be harmful to the viewer’s light,
thus reducing the contrast of the image.
Figure created with biorender.com
Applications of the Fluorescent Microscope
 Used in the visualization of bacterial agents such as Mycobacterium
tuberculosis.
 Used to identify specific antibodies produced against bacterial
antigens/pathogens in immunofluorescence techniques by labeling the
antibodies with fluorochromes.
 Used in ecological studies to identify and observe microorganisms
labeled by the fluorochromes
 It can also be used to differentiate between dead and live bacteria by
the color they emit when treated with special stains
Besides the above-discussed microscopes, there is one not commonly
used microscope known as the Differential Interference Contrast
Microscopy. It is very similar to the phase-contrast microscope whereby
the images are formed from the variations in the light either deviated and
or undeviated. The difference is, here two beams of light are emitted to
the specimen and focused by a prism. One beam passes through the
prism to the specimen while another passes through the glass slide clear
area without the specimen. The two beams then combine and interfere
with each other to form an image. It can be used to view cell structures
such as endospores, bacterial cell walls, nuclei and granules for unstained
specimens.
References
1. Microbiology by Laning M. Prescott, 5th Edition
2. https://science.umd.edu/CBMG/faculty/wolniak/wolniakmicro.html
3. https://en.wikipedia.org/wiki/Bright-field_microscopy
4. https://sciencing.com/calculate-magnification-light-microscope-
7558311.html
5. https://www.microscopemaster.com/brightfield-microscopy.html
6. https://en.wikipedia.org/wiki/Phase-contrast_microscopy
7. https://www.microscopyu.com/techniques/phase-
contrast/introduction-to-phase-contrast-microscopy
8. https://www.olympus-lifescience.com/en/microscope-
resource/primer/techniques/phasecontrast/phase/
9. https://laboratoryinfo.com/types-of-microscopes/