INTRODUCTION

MICROSCOPE
A microscope is an instrument used to view objects that are smaller than the human
eye.

Types of Microscopes:

1. Optical Microscopes (light microscopes)

• Compound Microscopes
• Dissection Microscopes (Stereo Microscopes)

2. Electron Microscope
• Transmission electron microscope
• Scanning electron microscope

3. Digital Microscope

Microscope Viewing Head Categories:

1. Monocular
• Viewing heads only use one eyepiece when viewing a specimen, microscopes
with this type of head are light weight and inexpensive.

2. Binocular
• These viewing heads have two eyepieces and are more comfortable for the
observer they are the most common choice of microscopes.
 3. Trinocular
• These viewing heads contain 3 eyepieces, two can be used by one user and
the third simultaneously by another person or by a camera which can be fitted
into the microscope, microscopes with this type of head are more expensive.

Main Parts and attachments of a Microscope:

1. Eyepiece Lens:
• The lens at the top of the microscope that you look through. They are
usually10X or 15X power.

2. Tube:
• Connects the eyepiece to the objective lenses.

3. Arm:
• Supports the tube and connects it to the base.

4. Base:
• The bottom of the microscope, used for support.

5. Illuminator:
• A steady light source (110 volts) used in place of a mirror. If the microscope
has a mirror, it is used to reflect light from an external light source up through
the bottom of the stage.

6. Stage:
• The flat platform where you place your slides or specimens in case of a Stereo
Microscope. Stage clips hold the slides in place, Depressions hold Specimens
in place. If the microscope has a mechanical stage, the user will be able to
move the slide around by turning two knobs. One moves it left and right, the
other moves it up and down.
 7. Revolving Nosepiece or Turret:
• This is the part that holds two or more objective lenses and can be rotated to
easily change the magnification power of the telescope.

8. Objective Lenses:
• Usually you will find 2 - 4 objective lenses on a microscope. They normally
consist of 4X, 10X, 40X and 100X powers. The shortest objective lens is the
lowest power, and the longest one is the lens with the greatest power. Lenses
are color coded, and some standards are interchangeable between
microscopes. The high-power objective lenses are retractable (i.e.40XR). This
means that if they hit a slide, the end of the lens will push in (spring loaded)
thereby protecting the lens and the slide.

9. Rack Stop:
• This is an adjustment that determines how close the objective lens can get to
the slide. It is set at the factory and keeps the user from cranking the high-
power objective lens down into the slide and breaking it.

10. Condenser Lens:

• The purpose of the condenser lens is to focus the light onto the specimen.
Condenser lenses are most useful at the highest powers (400X and above).
Microscopes within stage condenser lenses render a sharper image than those
with no condenser lens. If your microscope has a maximum power of 400X,
you will get the maximum benefit by using a condenser lens rated at 0.65 NA
(Numerical Aperture) or greater

11. Diaphragm or Iris:

• Many microscopes have a rotating disk under the stage. This diaphragm has
different sized holes and is used to vary the intensity and size of the cone of
light that is projected upward into the slide. There is no set rule regarding
 which setting to use for a particular power. Rather, the setting is a function of
the transparency of the specimen, the degree of contrast desired and the
particular objective lens in use.

12. Eyepiece diopter setting:

• Compensates for focusing differences between the eyes, it is very important
this is set correctly, in order to prevent eye strain.

13. Focus knob:

• Moves the head of the microscope up and down to bring the object sharply
into view.

14. Lighting (Stereo Microscopes):

• Top lighting shines down and reflects off opaque or solid specimens; bottom
lighting shines up through transparent objects. Not all microscopes will have
both.

15. Stage plate (Stereo Microscopes):

• This is where the specimen is placed for viewing located directly under the
objective lens. Some stereo microscopes have reversible black and white stage
plates to provide appropriate contrast with the object being viewed.

theory
 1. Optical Microscopes:

• Optical microscopes contain one or more lenses that produce a large image of
an object (applied engineering terms) placed in its focal plane, that object’s
image is magnified by projecting a beam of light through the object
(specimen), this beam of light is then refracted through the various lenses
within the microscope to focus the light into the eye where it is then processed
by the brain.

How a Multiple Optical Microscope works;

• The condenser lens focuses the beam of light from the light source onto the
sample (object) and then that light which contains the image goes through the
objective lenses which might be of magnification10x, 40x, 1000x to a projector
lens where it is viewed by the person observing the specimen (object).
➢ Compound Microscopes:
• A Compound Microscope Invented in 1590 by Zacharias Janssen and Dutch
Spectacle maker is a microscope fitted with two or more convex lenses
allowing them to have higher resolutions and greater magnification; these
microscopes are common in science laboratories and medical research
because of their ability to produce detailed images of micro-organisms, cells
and tissues.
→ How it works:
• The compound microscope is made up of convex lenses fitted at either end of
a hollow tube, this tube is fitted on an adjustable rotary nosepiece under which
is an adjustable stage which specimen slides are placed or fitted for
observation though the lenses, the stage has a hole or in it through which a
light source can illuminate the specimen to be observed. This light source can
either be a mirror reflecting natural light or a lamp in the microscope base as
the illuminating beam passes though the stage’s window (the hole) and
through the specimen the light brightens around the specimen making it
standout in contrast. The level or contrast is controlled by controlling the
amount of illumination light which goes through the specimen a brighter or
dimmer effect is achieved by opening or closing an iris diaphragm under the
stage or by adjusting the height of the lamp.
 • The upper lenses of the compound microscope closest to the observer’s eyes
are the ocular lenses or “ eyepiece”, the lower lenses closer to the object
(specimen) are the objective lenses, the rotary nose-piece may contain three or
four different objective lenses, to select the magnification required, the
nosepiece which contains the objective lenses is rotated to select the most
suitable magnification for the particular object (specimen), the four objective
lenses fitted in this type of microscopes are:
• The scanning power objective (magnification of 4x)
• Low power objective (magnification of 10x)
• High dry objective (*magnification of 40x)
• Oil immersion objective (magnification of 100x)

The ocular lens (eyepiece) usually has a magnification of 10x

To obtain the total magnification factor the eyepiece magnification is multiplied by
the objective magnification, so therefore if the “High dry objective” which has a
magnification of 40x is coupled with the eyepiece which usually has a magnification
of 10x then the total magnification of the specimen will be 400x so the specimen will
be viewed at 400x its actual size.

→ Image formation
• When the object is in focus the objective lenses for a real, inverted image of
the object (specimen) at a point inside the principle focus of the ocular lenses
(eyepiece), the eyepiece then treat this inverted image as the object and
produce a magnified upright image of the specimen.

Compound Microscope:
➢ Dissection Microscopes (Stereo Microscopes):
• These types of microscopes also called Stereo Microscopes are commonly
used in the lab or for examining archaeological artifacts and geological
samples, these microscopes have two eyepieces and must have two eyepieces
to be considered a Stereo Microscope and sometimes have a third lens for the
purpose of taking video or photographs of the specimen being worked with.
• They have two objective lenses allowing the observer to view the specimen at
two slightly different angles making the specimen look three dimensional (3D).
These types of microscopes have lower magnification power than compound
microscopes generally below 100x however there are types of Stereo
Microscopes which allow the user to assemble the stand objectives, eyespieces
and lighting device (source) to suit their need. In most cases the Stereo
Microscope have a greater distance between the objective and the stage than
that of a Compound Microscope, the stage is usually large and may have a
depression for securing specimens allowing them to be worked with or
dissected while it is being viewed (hence the name Dissection Microscope).
The type of illumination these microscopes uses are: LED ring light, fluorescent
bulbs or fiber optics. However most Stereo Microscopes work on natural light
 which is reflected off of the object rather than transmitted light coming from
beneath the stage.

2. Electron Microscopes:

• Electron Microscopes unlike optical microscopes use a beam of electrons to

illuminate the specimen so the image can be produced the image is formed by
a stream of electrons emitted by the electron source and accelerated toward
the object (specimen) by use of a positive electrical potential. The stream of
electrons is condensed and focused using metal apertures and magnetic
lenses into a focused monochromatic beam which is focused onto the object
(specimen) using a magnetic lens. The beam of electrons is affected by
interactions that occur within the object (specimen) and is then detected and
formed into an image that the observer can view.

• One advantage that Electron Microscopes have over optical microscopes is its
level of magnification, most optical microscopes can magnify up to 2000x (Two
Thousand Times) whereas modern Electron Microscopes can magnify an
image up 2million Times.

Types of Electron Microscopes:

1. Transmission Electron Microscopes and Scanning Electron Microscopes:
 • These types of microscopes work the same way as Electron Microscopes except the electron
source is an electron gun that emits the high voltage electrons to create an image.

• Transmission Electron Microscopes give a 2D detailed image of the specimen.

• Scanning Electron Microscopes give a 3D image. They also capture the image
of the specimen.

3. Digital Microscope:

• These microscopes use charged coupling device (CCD) sensors and no longer
rely on optical lenses alone, they have incorporated light sensitive pixels to
capture images which when compiled by a computer software allows the
image to be magnified and viewed in 2D, 3D and Real time, they are built on
the same basis as optical microscopes where a specimen is placed on the
stage beneath an objective lens, a light source emits light to make the sample
easier to see, the objective lens magnifies the specimen and it is brought up
into focus within the tube of the microscope, unlike in traditional microscopes
the Digital Microscope has no eyepiece instead it has a screen where the
image is viewed.

• These Microscopes can also most times connect to a computer via USB so that
the images the capture can be stored on the computer’s hard drive for later
viewing or inspection, the images can also be displayed on a monitor or
 projected by the use of specialized software so many people can see the
specimen while its being worked with.

• Digital Microscopes are dependent on the optics of the objective lens for
much of their magnification. The resolution of the camera and the monitor size
determine how much magnification is possible with a Digital Microscope
usually up to 1000x. This type of microscope does not necessarily include an
eyepiece, it may be possible to capture images with far greater optical
magnification. Also, images maybe further magnified digitally, especially with a
high-resolution camera.