Microscope

Introduction of Microscope
o A microscope is an optical instrument used to
magnify and observe small objects, samples or
structures that are not visible to the naked eye.

o It produces an enlarged image of the object, allowing

for detailed study and examination.

o Microscopy is the science that deals with the use of

microscopes and the interpretation of their magnified
images.
Continue…

o Microscopes use a combination of lenses, mirrors and

other optical components to:
1) Collect and focus light from the object
2) Magnify the image
3) Produce a clear and resolve image

The use of these microscopes has provided with the

microbiologists new insights into the
microorganisms and their role in the environment
and their relationship with other forms of life on
our planet.
History of Microscope
Types of Microscope
Bright field microscope

Dark field microscope

Phase contrast microscope

Fluorescence microscope

Electron microscope
Bright field Microscope
A microscope that allows light rays to pass
directly through object to the eye without
being deflected by an intervening opaque
plate in the condenser.
It is usually known as compound light
microscope, because it possesses at least
two lens systems.
Components of a microscope
Eyepiece
Display the image formed by the objective lens for the
viewer
Nosepiece

Revolving circular structure

containing the objectives.

Makes it possible to move

them into position over slide.
 Objectives

Lens that receives light from

object to form a magnified image
that you view through the
eyepiece
Low power (short)
High power (long)
Oil immersion
Arm and Base

These components of microscope

are known as basic frame
structure of microscope, because Arm
all other parts are attached them.

Arm of the microscope is also

used for carrying

Base of the microscope is also

support to the microscope.

base
 Stage
The stage is the horizontal plate
form that support the glass slide.
It has a clamped device, the
mechanical stage, which is used
for holding and moving the slide
around on the stage with the help
of mechanical stage control.
Stage clips

Used to hold the slide on

the stage
Diaphragm

This lens system is the condenser, which is located

under the stage.

It collect and brings the light rays from the illuminator to a

common focus on slide being studied.

The condenser can be moved up and down by a knob

under the stage.

It contains an attachment called diaphragm that controls

the amount of light that passes up through the bottom of
the stage to the slide.
A diaphragm lever is present to open or close
diaphragm.
Focusing knob

o The focusing knob are located on

the side of the microscope and are
use for bringing objects into focus
when studying an object on a
slide.
o There are two types of focusing
knob: course adjustment that
brings an approximate focus and
fine adjustment that brings the
object into perfect focus.
 Light source

In the base of the most microscope

is positioned an illuminator.
Provides a steady stream of light
that passes up through the bottom
of the stage
 Magnification
Magnification is the power of a microscope to enlarge
the objects.
It is obtained in the compound microscope by the
interaction between light waves, ocular and objective.
The object first magnifies the specimen and produce a
real image.
The real image projected through the microscope to
the ocular lens, which magnifies the real image and
produce an image seen by the observer and called
virtual image.
Resolution
The resolving power of a microscope refers to its
ability to distinguish between two closely spaced
objects as separate entities.
It's a measure of how much detail can be observed in
the specimen being viewed.
The resolving power is determined by several factors,
including the wavelength of light (or other radiation)
used, the numerical aperture of the optical system,
and the quality of the lenses.
 Dark field microscope
Dark field microscope is an important
tool for the observing transparent
lining organisms particularly those
which have a diameter less then the
resolution of the bright field
microscope
It is widely used for the identification
of spirochetes in the exudates from a
syphilitic lesion.
The cells in the specimen become
brightly illuminated against a dark
background because of light scattering
Dark field photomicrograph of Treponema pallidum, the
cells appear as illuminated structures against a dark
background
Phase contrast microscope
This microscope is able to differentiate transparent
protoplasmic structures without staining and killing
them.
The first phase contrast microscope was developed in
1933 by Fredrick Zernike and was originally referred to
as the Zernike microscope
It is the instrument for choice for studying living
protozoa and other types of transparent cells.
This type of microscope consists of special condenser
and objectives that enable one to increase the
contrast between the transparent component in the
cell by exploiting differences in their densities.
Protozoa photoghraphed through a phase contrast
microscope appear shiny.
The bright area are less dense then the darker area inside
the cell.
 Fluorescent microscope
This is a unique instrument that is
used to observe cells that have been
stained with special compounds
that possess the quality of
fluorescence
Such compounds are called
fluorochromes.
Auramine O, acridine orange, and
fluorescein are well known
fluorochromes.
 Electron microscope
o An electron microscope is a powerful scientific instrument
used to visualize objects at a very high resolution by using a
beam of electrons rather than light to illuminate the
specimen.
o This type of microscope can achieve much higher
magnification and resolution compared to traditional light
microscopes, allowing researchers to study the fine details
of objects, such as the structure of cells, viruses,
nanoparticles, and crystalline structures.
o The only drawback to this microscope is that living objects
can not be examined.
o There are two types of electron microscopes:
o transmission electron microscopes (TEM)
o scanning electron microscopes (SEM)
Rules of using a microscope

Always carry with both hands

Only use lens paper for cleaning

Do not force knobs

Always store covered

Be careful of the cords

 How to use a microscope

Place the slide on the stage

Use stage clips to secure slide

Adjust nosepiece to lowest setting

(Lowest = shortest objective)

Look into eyepiece

Use coarse focus knob

Troubleshooting
Occasionally you may have trouble with working your microscope. Here are
some common problems and solutions.

1. Image is too dark!

Adjust the diaphragm, make sure your light is on.

2. There's a spot in my viewing field, even when I move the slide the spot stays in
the same place!
Your lens is dirty. Use lens paper, and only lens paper to carefully clean the
objective and ocular lens. The ocular lens can be removed to clean the inside.
The spot is probably a spec of dust.

3. I can't see anything under high power!

Remember the steps, if you can't focus under scanning and then low power, you
won't be able to focus anything under high power. Start at scanning and walk
through the steps again.

4. Only half of my viewing field is lit, it looks like there's a half-moon in there!
You probably don't have your objective fully clicked into place..
Application of Microscope
Biology
Studying cells, tissue, microorganisms
Medicine
Diagnosing diseases, examining tissues
Materials science
Analyzing materials, structures
Research
Scientific investigation, discovery
Education
Teaching, learning
Practice Labeling the Parts