BIO 101 (BASIC PRINCIPLES OF BIOLOGY)

TOPIC:

MICROSCOPY

Section taken by: Dr Bola Oyefolu

Microbiology Dept.,
Faculty of Science, LASU, Ojo. 1
 DEFINITION OF MICROSCOPE

• Microscope is an
instrument which provides
an enlarged image of
minute objects such as;
sub cellular structures, and
many more that are
generally not visible to the
naked eyes. The complexity of microscopes has
The word “microscope” is however since its invention, increased
formed of two Greek to many folds from simple lens to
words: “micros”- small and complex scanning electron microscope
‘skipein’-to look. 
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 CONCEPT OF MICROSCOPY
Microscopy is the science
of investigating small
objects and structures
using such an instrument.
 microscopy, the microscope
In
must accomplish three tasks. It
must:
1.Produce a magnified image of
the specimen,
2.Separate the details in the
image and,
3.Render the details visible to the
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human eye or camera.
 CONCEPT OF MICROSCOPY
SCALE:

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THE HISTORY OF MICROSCOPE

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 HISTORY OF MICROSCOPE Contd.
PRE 1600:
Zoocharia Jansen in 1590 and his
brother Hans in 1595 used second
lens that enlarged imaged formed
by first lens by 50-100X.

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 HISTORY OF MICROSCOPE Contd.
PRE 1660:
In 11th century , the Arab
Alhazan described the use
and characteristics of glass
lenses
Roger Bacon was familiar
with lenses and eye glasses ,
however weren’t invented until
late 1200s 

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 HISTORY OF MICROSCOPE Contd.
In the early & mid 1660s:
In 1608 Telescope was invented ,
with Galileo improving upon it with
his own models
Around 1600 microscope was
invented possibly by Hans and
Zacharias Jansen. But lens quality
was poor.
The first known image of a
microscope is a drawing by Isaac
Beeckman in 1631.
FIRST BIG MICROSCOPE (with
200X maximum magnification):
came in 1665 when Robert Hooke
published the book
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“MICROGRAPHIA “
 HISTORY OF MICROSCOPE Contd.
In the late 1660s:
Anton van Leeuwenhoek
began to grind his own lens and
make simple microscope.
He discovered nematode and
rotifers.
.

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 HISTORY OF MICROSCOPE Contd.
In the late 1660s:
Anton van Leeuwenhoek in
1673, provided improved
microscope and was first to
observe unicellular animal. He
is called “Father Of
Microbiology”.

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 HISTORY OF MICROSCOPE Contd.
Between 1700 & 1800s:
 Not much change in basic
microscope butt certain
problems were resolved n this
era like color distortion and poor
image resolution.
 Ernst Abbe : A German
Physicist opined that oil
emersion lense can prevent
length distortion .
.

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 HISTORY OF MICROSCOPE Contd.
1900 till NOW!:
In 1931, a German scientist,
Ernst Ruska invented electron
microscope which can magnify
as much as million times.
The only drawback is that
living cell can not be viewed by
using it.

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 TYPES OF MICROSCOPE

 Basically there are two types of microscopes:

1. Light Microscope
2. Electron Microscope

 Further classification in these basic microscopes are

present. Other microscopes also present such as:
- Phase contrast Microscopes
- Fluorescent Microscope
- Bright field AND Dark field Microscope
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 TYPES OF MICROSCOPE

 Most commonly used microscope.

 Handy in use.
 LM uses light source for illumination of specimen.
 Generally used light sources include sunlight, UV
light, laser light, LEDs.
 Types- Simple dissecting microscope, compound
microscope, stereomicroscopes.

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 TYPES OF MICROSCOPE contd.

1. Light Microscope (LM):

 Light Microscope uses sunlight or artificial light. It is the
most commonly used microscope.
Handy in use.
LM uses light source for illumination of specimen.
Generally used light sources include sunlight, UV light, laser
light, LEDs.
LM includes: Simple dissecting microscope, compound
microscope, stereomicroscopes. The various other types of
light microscope are:

A.Bright field microscope.

B.Dark field microscope.
C.Phase contrast microscope.
D.Fluorescence microscope.
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 TYPES OF MICROSCOPE contd.

2. Simple Microscope:
 Consist of Biconvex lense.
Can be moved up and
down by adjustment.
Object is placed on a
platform.
Light is focused by
concave mirror.

A Simple Dissecting Microscope

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 TYPES OF MICROSCOPE contd.

3. Compound Microscope:
 Has 2-sets of lenses.
An Objective lens of
short aperture and focal
length
Another set of lens of
larger aperture and focal
length facing eye and
known as “Eye piece”

Compound Microscopes
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 TYPES OF MICROSCOPE contd.
4. Stereo-Microscope:
This microscope allows for binocular
(two eyes) viewing of larger specimens.
This is otherwise known as Dissecting
Microscope and serves a different
purposes such as:
Can be used for thicker specimens
It produces a three dimensional (3-D)
visualization of the sample being
examined.
 It is used for dissections.
Its used to study the surfaces of solid
specimens or to carryout close work such
as: sorting, micro-surgery, watch-making,
small circuit board manufacture or
inspection, insects and leaves and the
likes.
 Usually magnifies 10x to 20x.
Stereo-microscopes 18
 TYPES OF MICROSCOPE contd.

5. Interference
Microscope:
 It is used for
quantitative studies
of macromolecules
of the cell
components

Interference Microscopes 19
 TYPES OF MICROSCOPE contd.

Metallurgical IM

6. Inverted Microscope (IM):

It has both the light source and condenser set up high
above the stage and pointing down toward the stage.
The objectives and objective turret are located beneath
the stage pointing up using reflecting objectives.
The two basic types of inverted microscopes include
biological inverted (BIM) and metallurgical inverted (MIM)
microscopes. Biological IM
IM is used in micro-manipulation applications where
space above the specimen is required for manipulator
mechanisms and the microtools they hold, and in
metallurgical applications where polished samples can be
placed on top of the stage and viewed from underneath.
The BIM is used to observe cell morphology.

Inverted Microscopes
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 PRINCIPLES OF MICROSCOPY

IMAGES AS SEEN BY DIFFERENT MICROSCOPES 21

 TYPES OF MICROSCOPE contd.

7. Electron microscope (EM or e-microscope):

It uses electron. The various type of e-microscopes
are:
1. Transmission e-microscope.
2. Scanning e-microscope

In 1931, a German scientist, Ernst Ruska invented

electron microscope which can magnify as much as
million times.
The only drawback is that living cell can not be viewed
by using it.
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 PARTS OF MICROSCOPE

Parts of simple Microscope

Basically, the microscope is divided into two(2)major parts:
1.Mechanical parts
2.Optical parts

Mechanical Part:
1.This is composed of the body frame which could be
unscrewed and serviced.

2.Optical Part: These parts are involved in passing the light

through the object (specimen) and magnifying its size.

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 PARTS OF MICROSCOPE
Parts of simple Microscope
The components of the optical parts are as follows:
Mirror:
-A Plano-convex mirror is fitted below the stage to the vertical
rod by means of a frame.
-It focuses the surrounding light on the object to be observed.
Lens:
-A biconvex lens is fitted above the stage, to the vertical rod, by
means of a frame.
-It magnifies the size of the object and the enlarged virtual
image formed is observed by keeping the eye above it.
-For proper focusing, the lens can be moved up and down by
the frame.
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 PARTS OF MICROSCOPE
Parts of simple Microscope
Eyepiece: The lens the viewer looks through to see the
specimen. The eyepiece usually contains a 10X or 15X power
lens.
DiopterAdjustment: Useful as a means to change focus on one
eyepiece so as to correct for any difference in vision between
your two eyes.
Body tube (Head): The body tube connects the eyepiece to the
objective lenses.
Arm: The arm connects the body tube to the base of the
microscope.
Coarse adjustment: Brings the specimen into general focus.
Fine adjustment: Fine tunes the focus and increases the detail
of the specimen. 25
PARTS OF MICROSCOPE
Parts of a Simple Microscope

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 PARTS OF MICROSCOPE
How Does The Microscope Work
 All of the parts of a microscope work together.

 The light from the illuminator passes through the aperture,

through the slide, and through the objective lens, where the
image of the specimen is magnified.

 The then magnified image continues up through the body tube

of the microscope to the eyepiece, which further magnifies the
image the viewer then sees.

 Learning to use and adjust your compound microscope is the

next important step.

 It's also imperative to know and understand the best practices of

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cleaning your microscope.
PARTS OF MICROSCOPE

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PARTS OF MICROSCOPE

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PARTS OF MICROSCOPE

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PARTS OF MICROSCOPE

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PARTS OF MICROSCOPE

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PARTS OF MICROSCOPE

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PARTS OF MICROSCOPE

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PARTS OF MICROSCOPE

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 MAGNIFICATION IN MICROSCOPY

MAGNIFICATION
Your microscope has 3 magnifications:
- Scanning,
-Low and
-High.
-Each objective will have written the
magnification.
-In addition to this, the ocular lens
(eyepiece) has a magnification.
-The total magnification is the ocular x
objective

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 PRINCIPLES OF MICROSCOPY
LIGHT MICROSCOPY:
Microscopes are of great importance
in the study of microorganisms and
biomolecules.
Light microscopes are simplest of all
microscopes.
In light microscopy, light typically
passes through a specimen and then
through a series of magnifying lenses
Light microscopes use lenses to
bend and focus light rays to produce
enlarged images of small objects.
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 PRINCIPLES OF MICROSCOPY
Workings of Light Microscope (LM):
The specimen is mounted on slide and positioned
in specimen stage.
Beam of light is focused on specimen by
condenser.
Objective lens picks up light transmitted by
specimen and produce first magnified image.
This image is further magnified by eyepiece lens.
Eyepiece only magnifies image and brings no
change in resolution.
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 TYPES OF MICROSCOPE contd.

APPLICATIONS OF LIGHT MICROSCOPE (LM)

1. LM is used to study preserved minute specimen.

2. Used to study activities inside the cell.

3. Used in identifying macromolecules of cell.

4. Used in Medical diagnosis.

5. Used in histopathological studies.

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 PRINCIPLES OF MICROSCOPY
WORKINGS OF COMPOUND MICROSCOPE:
 Light is transmitted and focused by
mirror and condenser. Magnification
 Focused light illuminate the object or Total magnification of
specimen. specimen is by
 The refracted light is collected by an multiplying the
objective lens
objective where primary image of the magnification power by
object is formed, it is real, inverted the ocular lens
enlarged image of the object. magnification power.
 The eyepiece further magnifies this Low power X10,
primary image into virtual, erect High power X40 and Oil
enlarged image, this is the final image Immersion X100
that lies above the stage.
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 PRINCIPLES OF MICROSCOPY
WORKINGS OF COMPOUND MICROSCOPE contd:
Resolution:
Is also called resolving power of image.
This mean, the ability to distinguish that two
objects are separate and not one.
Resolving power of a microscope is determined
by the wave length of light entering the objective
lens.
A general principle of microscopy is that the
shorter the wave length of light used in the
instrument, the greater the resolution.
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 PRINCIPLES OF MICROSCOPY
WORKINGS OF COMPOUND MICROSCOPE contd:
Oil Immersion:
The white light used in a compound light microscope
has relatively long wave length and cannot resolve
structures smaller than about 0.2 µm.
Immersion oil is placed between the glass and
objective lens. The immersion oil has the same
refractive index as glass of the microscope.
The oil enhances the resolution by preventing light
rays from dispersing and changing wave length after
passing through the specimens.
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 PRINCIPLES OF MICROSCOPY
WORKINGS OF COMPOUND MICROSCOPE contd:
Applications:

Observation of morphology of microorganisms.

Detection of cell structures.
Observation of intracellular structures.
Observation of motility.
Measurement of size.
Observation of blood smears.

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 PRINCIPLES OF MICROSCOPY
WORKINGS OF BRIGHT-FIELD MICROSCOPE:
A. Bright-Field Microscopy:
The ordinary microscope is also referred to as Bright-
Field microscope.
It forms dark image against bright background.
The useful magnification of Light microscope is limited
by its resolving power.
The resolving power in limited by wavelength of
illuminating beam.
Resolution is determine by certain physical parameters
like wave length of light and light generating power of the
objective & condenser lens. 44
 PRINCIPLES OF MICROSCOPY
WORKINGS OF BRIGHT-FIELD MICROSCOPE:

In bright field, microscope field appears bright

whereas microorganism appears dark as they
absorb light.

Normally micro-organism do not absorb light but

absorbing ability increases due to staining.

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 PRINCIPLES OF MICROSCOPY
HOW IMAGE IS FORMED IN BRIGHT-FIELD MICROSCOPE:
 Image is created by
objective and ocular lenses
working together.
 Light from illuminated
specimen is focused by the
objective lens creating
enlarged image within the
microscope.
 The ocular lens further
modifies the primary image.
 Total magnification is
calculated by magnification
by objective multiply by
magnification by eyepiece.
 E.g. : 45x X 10x =450x

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PRINCIPLES OF MICROSCOPY

IMAGE FORMED IN BRIGHT-FIELD MICROSCOPE:

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 PRINCIPLES OF MICROSCOPY

ADVANTAGES OF BRIGHT-FIELD MICROSCOPE:

 Bright field compound microscopes are commonly used

to view live and immobile specimens such as bacteria,
cells, and tissues.

 For transparent or colorless specimens, however, it is

important that they be stained first so that they can be
properly viewed under this type of a microscope.

 Staining is achieved with the use of a chemical dye. By

applying it, the specimen would be able to adapt the color
of the dye. Therefore, the light won’t simply pass through
the body of the specimen showing nothing on the
microscope’s view field 48
 PRINCIPLES OF MICROSCOPY

WORKINGS OF DARK-FIELD MICROSCOPE:

B. Dark-Field Microscopy:
Dark field microscopy is frequently performed on the
same microscope on which bright-field microscopy is
performed.
Instead of the normal condenser that contains an opaque
disk.
The disk blocks light that would enter the objective lens
directly.
Only light that has been reflected or refracted by the
specimen forms the image.
The field surrounding specimen appears dark while the
object brightly illuminated 49
 PRINCIPLES OF MICROSCOPY

WORKINGS OF DARK-FIELD MICROSCOPE:

In dark field microscope, a dark background

is established against a brightly illuminated
object.

Dark field microscopy requires additional

dark field condenser and dark field object
lens.

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 PRINCIPLES OF MICROSCOPY
HOW IMAGE IS FORMED IN DARK-FIELD MICROSCOPE:

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 PRINCIPLES OF MICROSCOPY

ADVANTAGES OF DARK-FIELD MICROSCOPE:

 The advantage of darkfield microscopy also becomes its

disadvantage: not only the specimen, but dust and other
particles scatter the light and are easily observed.
 For example, not only the cheek cells but the bacteria in saliva
are evident.
 The dark field microscopes divert illumination and light rays
thus, making the details of the specimen appear luminous.
 Dark field light microscopes provide good results, especially
through the examination of live blood samples.
 It can yield high magnifications of living bacteria and low
magnifications of the tissues and cells of certain organisms.
 Certain bacteria and fungi can be studied with the use of dark
field microscopes.
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 PRINCIPLES OF MICROSCOPY
WORKINGS OF PHASE CONTRAST MICROSCOPE:

C. PHASE CONTRAST Microscopy:

Phase contrast microscope was developed by Fritz Zernike,
and was awarded Nobel prize in Physics in 1953.
By this, microscopy organism can be seen alive, without
staining.
It requires additional specialized structure annular
diaphragm and phase contrast ring.
The images differentiates in refractive index of cellular
structure.
Light passes through thicker parts of cell is held up relative
to the light that passes through thinner parts of cytoplasm.

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 PRINCIPLES OF MICROSCOPY
WORKINGS OF PHASE CONTRAST MICROSCOPE:

C. PHASE CONTRAST Microscopy:

The principle of phase-contrast microscopy is based on the
wave nature of light rays and the fact that light rays can be in
phase or out of phase.

In a phase-contrast microscope, one set of light rays comes

directly from the light sources.

The other set comes from light that is reflected or diffracted

from a particular structure in the specimen (diffraction is the
scattering of light rays-direct rays and reflected or diffracted
rays are brought together) together).
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 PRINCIPLES OF MICROSCOPY
WORKINGS OF PHASE CONTRAST MICROSCOPE:

PHASE CONTRAST Microscopy contd.:

Both combined rays form an image of the specimen on
the ocular lens, containing areas that are relatively light
(in-phase), through shades of gray, to black(out phase).

Through the use of annular lens, the differences in phase

are amplified so that in-phase light appears brighter than
out-of-phase.

It is used to study the behaviour of living cells and;

To observe nuclear and cytoplasmic changes.

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 PRINCIPLES OF MICROSCOPY
HOW IMAGE IS FORMED IN PHASE CONTRAST MICROSCOPE:

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 PRINCIPLES OF MICROSCOPY
HOW IMAGE IS FORMED IN PHASE CONTRAST MICROSCOPE:

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 PRINCIPLES OF MICROSCOPY
WORKINGS OF PHASE CONTRAST MICROSCOPE:
Applications:
Observation of morphology of microorganisms.
Detection of cell structures.
Observation of intracellular structures.
Observation of motility.
Measurement of size.
Observation of blood smears.
To study unstained living cells.
Detailed examination of internal structures In living
microorganism living.
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 PRINCIPLES OF MICROSCOPY
WORKINGS OF PHASE CONTRAST MICROSCOPE:
Applications contd.:
To study flagellar movements and motility of bacteria
and protozoans. bacteria and protozoans.
To study intestinal and other living protozoa such as
amoeba and trichomonas.
To examine fungi grown in culture
Phase contrast microscopy is used in study of living
cells and tissues.
Microbes and parasites can also be studied.
Useful in observing cells cultured in vitro during
mitosis. 59
 PRINCIPLES OF MICROSCOPY

FLUORESCENCE MICROSCOPE
 Popularly used to achieve high labeling of cellular
compartments.

 This microscope additionally requires an

excitation filter, a barrier and a dichromatic
mirror, fluorescent stain.

 A specific wavelength of light is used to excite

fluorescent molecule in specimen.

 Light of higher wavelength is then imaged. 60

 PRINCIPLES OF MICROSCOPY
FLUORESCENCE MICROSCOPE
WHY FLUORESCENCE MICROSCOPE?

 In all types of microscopes, cell

constituents are not distinguishable,
although staining does , but not totally.
 In fluorescent microscopy, various
fluorescent dyes are used which gives
property of fluorescence to only specific
part of the cell and hence it can be
focused.. 61
 PRINCIPLES OF MICROSCOPY
THE WORKINGS OF FLUORESCENCE MICROSCOPE
 When certain compounds are illuminated
with high energy light, they then emit light
of a different, lower frequency.
 This effect is known as fluorescence.
 Often specimens show their own
characteristic auto-fluorescence image,
based on their chemical makeup.

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 PRINCIPLES OF MICROSCOPY
THE WORKINGS OF FLUORESCENCE MICROSCOPE
 Many different fluorescent dyes can be
used to stain different structures or
chemical compounds.
 One particularly powerful method is the
combination of antibodies coupled to a
fluorochrome as in immunostaining.
 Examples of commonly used
fluorochromes are fluorescein or
rhodamine. 63
 PRINCIPLES OF MICROSCOPY
THE WORKINGS OF FLUORESCENCE MICROSCOPE

1. A component of interest in the specimen is specifically

labeled with a fluorescent molecule called a fluorophore
called a fluorophore
2. The specimen is illuminated with light of a specific
wavelength (or wavelengths) which is absorbed by the
fluorophores, causing them to emit longer wavelengths
of light (of a different color than the wavelengths of light
absorbed).
3. Typical components of a fluorescence microscope are the
light source (xenon arc lamp or mercury vapor lamp), the
excitation filter, the dichroic vapor lamp), the excitation
filter, the dichroic mirror and the emission filter.
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 PRINCIPLES OF MICROSCOPY
THE WORKINGS OF FLUORESCENCE MICROSCOPE

4. The illumination light is separated from the much

weaker emitted fluorescence through the use of an
emission filter.

5. The filters and the dichroic are chosen to match the

spectral excitation and emission characteristics of the
fluorophore used to label the specimen.

6.In this manner, a single fluorophore (color) is imaged

at a time. Multi-color images of several fluorophores
must be composed by combining combining several
single-color images.
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 PRINCIPLES OF MICROSCOPY
THE WORKINGS OF FLUORESCENCE MICROSCOPE

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PRINCIPLES OF MICROSCOPY

FLUORESCENCE MICROSCOPE 67
 PRINCIPLES OF MICROSCOPY

IMAGE AS SEEN BY FLUORESCENCE MICROSCOPE

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 PRINCIPLES OF MICROSCOPY

IMAGES AS SEEN BY FLUORESCENCE MICROSCOPE

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 PRINCIPLES OF MICROSCOPY
APPLICATIONS OF FLUORESCENCE MICROSCOPE
Fluorescence microscopy is a critical tool for:

Academic Research

Pharmaceutical Research,

Pathology,

Clinical Medicine.

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 PRINCIPLES OF MICROSCOPY

ELECTRON MICROSCOPE (EM)

INTRODUCTION
 German scientists Knoll and Ruska discovered electron
microscope.
 Electron microscopy (EM) is in some ways comparable to
light microscopy.
 Rather than using glass lenses, visible light, and the eye
to observe the specimen, the EM uses electromagnetic
lenses, electrons, and a fluorescent screen to produce the
magnified image.
 Electron beam is used as source of illumination.
 That image can be captured on photographic film to create
an electron photo-micrograph.
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 PRINCIPLES OF MICROSCOPY

ELECTRON MICROSCOPE (EM)

INTRODUCTION CONTD.
 The superior resolution of the EM is due to the
fact that electrons have a much shorter
wavelength than the photons of white light.
 The electron beam is focused by circular electron
magnets, which are analogous to the lenses in the
light microscope.
 The object which is held in the path of the beam
scatters the electrons and produces an image
which is focused on a fluorescent viewing screen.
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 PRINCIPLES OF MICROSCOPY

ELECTRON MICROSCOPE (EM)

INTRODUCTION CONTD.
 The wavelength of electrons used in EM is 0.005nm
as compared to 500nm with visible light i.e. about
100,000 times shorter than that of ordinary light.
 Theoretically, if conditions were identical in the
optical and EMs, the resolving power of the EM
should be 100,000 times (resolution down to 0.0025).
 However, the numerical aperture of an EM lens is
very small and does not approach the width of the
objective lens of an optical microscope.

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 PRINCIPLES OF MICROSCOPY

ELECTRON MICROSCOPE (EM)

INTRODUCTION CONTD.
 The electron beam is focused by circular electron
magnets, which are analogous to the lenses in the
light microscope.
 The object which is held in the path of the beam
scatters the electrons and produces an image
which is focused on a fluorescent viewing screen.

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 PRINCIPLES OF MICROSCOPY

ELECTRON MICROSCOPE (EM)

TYPES OF ELECTRON MICROSCOPE

1. Transmission Electron Microscope

2. Scanning Electron Microscope

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 PRINCIPLES OF MICROSCOPY

ELECTRON MICROSCOPE (EM)

TYPES OF ELECTRON MICROSCOPE
1. TRANSMISSION Electron Microscope (TEM)
In a Transmission e- microscope,
 Electrons pass through the specimen and scattered.
 Magnetic lenses focus the image onto a fluorescent
screen or photographic plate.
 Electron light pass directly through the specimen that has
been prepared by thin sectioning, freeze fracturing, or
freeze etching.
 TEM is used to observe internal and ultra-structures (fine
details) of cell structure.
 TEM Magnification is over 20million times. 76
 PRINCIPLES OF MICROSCOPY

TRANSMISSION ELECTRON MICROSCOPE (TEM) 77

 PRINCIPLES OF MICROSCOPY

TRANSMISSION ELECTRON MICROSCOPE (TEM) 78

PRINCIPLES OF MICROSCOPY

TRANSMISSION ELECTRON MICROSCOPE (TEM) 79

PRINCIPLES OF MICROSCOPY

IMAGES AS SEEN USING TEM 80

 PRINCIPLES OF MICROSCOPY

Cilia on Rabbit Lungs AS SEEN BY TEM MICROSCOPE 81

 PRINCIPLES OF MICROSCOPY

Chloroplast from a Tobacco Leaf AS SEEN BY TEM 82

PRINCIPLES OF MICROSCOPY

H1N1 Virus AS SEEN BY TEM 83

 PRINCIPLES OF MICROSCOPY
ELECTRON MICROSCOPE (EM)
TYPES OF ELECTRON MICROSCOPE
2. SCANNING Electron Microscope (SEM)
In a Scanning e- microscope,
Primary electron sweep across the specimen and knock
electrons from its surface.
The second electron is picked up by a collector, amplified,
and transmitted onto viewing screen or photographic plate.
It scan a beam of electrons back and forth over the surface
of a specimen producing three dimensional views of the
surfaces of whole microorganism.
SEM is use to stufy the surface of cell(s) and organisms.
SEM Magnification is 20,000 times.

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 PRINCIPLES OF MICROSCOPY

SCANNING ELECTRON MICROSCOPE (TEM) 85

 PRINCIPLES OF MICROSCOPY

SCANNING ELECTRON MICROSCOPE (TEM) 86

 PRINCIPLES OF MICROSCOPY

SCANNING ELECTRON MICROSCOPE (SEM) 87

 PRINCIPLES OF MICROSCOPY
ELECTRON MICROSCOPE (EM)
WORKINGS OF ELECTRON MICROSCOPE
 Beam of electron travel through column of
microscope in vacuum.
 Different electromagnetic lens focuses electrons
into thin beam.
 Beam travel through specimen. Some electrons
may get scattered while others transmitted (TEM),
hit fluorescent screen(detector) and forms image.
 In SEM, reflected electron from specimen forms
magnified image.

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PRINCIPLES OF MICROSCOPY

IMAGES AS SEEN USING SEM 89

 PRINCIPLES OF MICROSCOPY

IMAGES AS SEEN BY SCANNING ELECTRON MICROSCOPE (SEM) 90

 PRINCIPLES OF MICROSCOPY

Spirillum volutans --Electron micrograph showing

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individual flagella …AS SEEN BY SEM
 PRINCIPLES OF MICROSCOPY

Leptospira biflexa -- Electron micrograph showing

axial filament …AS SEEN BY SEM 92
PRINCIPLES OF MICROSCOPY

IMAGES AS SEEN USING SEM 93

 PRINCIPLES OF MICROSCOPY

HEAD OF DRAGON FLY AS SEEN BY SEM 94

PRINCIPLES OF MICROSCOPY

SPIDER AS SEEN BY SEM 95

 PRINCIPLES OF MICROSCOPY

HEAD OF BUTTERFLY AS SEEN BY SEM 96

 PRINCIPLES OF MICROSCOPY

A SPIDER THREAD AS SEEN BY SEM 97

PRINCIPLES OF MICROSCOPY

A FLEA AS SEEN BY SEM 98

 PRINCIPLES OF MICROSCOPY

A, B, C pollen grains: Scanning electron microscope D pollen grains:

Confocal Laser Scanning Microscope E pollen grains: Transmission
electron microscope F pollen grains: Light microscope G Mixed pollen
grains (bright field light microscope, stained) H pollen grains confocal
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laser scanning microscope
PRINCIPLES OF MICROSCOPY

A Dentist Drill AS SEEN BY SEM 100

PRINCIPLES OF MICROSCOPY

Toilet Paper AS SEEN BY SEM 101

PRINCIPLES OF MICROSCOPY

Hypodermic Needle AS SEEN BY SEM 102

PRINCIPLES OF MICROSCOPY

Velcro AS SEEN BY SEM 103

PRINCIPLES OF MICROSCOPY

Staple through a paper AS SEEN BY SEM 104

PRINCIPLES OF MICROSCOPY

Black Widow Spider Claw AS SEEN BY SEM 105

PRINCIPLES OF MICROSCOPY

PORCUPINE QUILL AS SEEN BY SEM 106

PRINCIPLES OF MICROSCOPY

MASCARA BRUSH AS SEEN BY SEM 107

PRINCIPLES OF MICROSCOPY

ANT AS SEEN BY SEM 108

PRINCIPLES OF MICROSCOPY

BLACK FLY AS SEEN BY SEM 109

PRINCIPLES OF MICROSCOPY

CAT FLY AS SEEN BY SEM 110

PRINCIPLES OF MICROSCOPY

MITE FEEDING AS SEEN BY SEM 111

PRINCIPLES OF MICROSCOPY

POLLEN GRAIN AS SEEN BY SEM 112

PRINCIPLES OF MICROSCOPY

ANT EYE AS SEEN BY SEM 113

PRINCIPLES OF MICROSCOPY

APHID ON A LEAF AS SEEN BY SEM 114

PRINCIPLES OF MICROSCOPY

EYE-LASHES AS SEEN BY SEM 115

PRINCIPLES OF MICROSCOPY

DOG- FLEA AS SEEN BY SEM 116

PRINCIPLES OF MICROSCOPY

VARIOUS VIRUSES AS SEEN BY SEM 117

PRINCIPLES OF MICROSCOPY

BACTERIA CELLS AS SEEN BY SEM 118

 PRINCIPLES OF MICROSCOPY

COMPARISON OF VIRUS AND BACTERIA CELLS AS SEEN BY SEM 119

 PRINCIPLES OF MICROSCOPY

COMPARISON OF EUGLENA IMAGES AS SEEN BY TEM AND SEM 120

 PRINCIPLES OF MICROSCOPY
ELECTRON MICROSCOPE (EM)
APPLICATIONS OF ELECTRON MICROSCOPE (EM)

1. EM is used to study the causes of disease.

2. Used to study 3D structure of cells .

3. Analysis of surface fracture or surface

contamination of cells.

4. Important part in production of silicon chips.

5. Used in industrial search centers.

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THANK YOU!!!

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