College of medicine
Anatomy department
Biology
1st stage
Microscope
Practical Biology
st
1 semester
Aysar M. Al- Atea
The light microscope
• Light microscope is a device can extend our ability to
see detail by 1000 times of ,
• so that we can see objects as small as 0.1 micrometer
(μm) or 100 nanometers (nm) in diameter.
• Without microscopes, our understanding of the
structures and functions of cells and tissues would be
severely limited.
PARTS OF A COMPOUND MICROSCOPE:
• Eyepiece/Ocular Lens:
• The eyepieces are the lenses you look through.
• The eyepiece of most binocular microscopes can be adjusted to
match the distance between the eyes of different observers
(interpupillary adjustment)
• Other microscopes may have different magnifications which are
stamped on the side of the eyepiece.
• The magnification of ocular lens are 10X.
• You may have to remove the eyepiece from its holder on the top
of microscope
• Eyepieces typically have a magnification between 5x & 30x.
Body Tube:
• The head part of microscope that connecting the light between the
objective lenses and the oculars
• The Light travels from the objectives through a series of magnifying
lenses to the body tube to the ocular.
• The oculars are held at an angle.
• The body tube contains a prisms that bends the light rays so that they
will pass through the oculars.
The eye tube angle in common
compound microscopes are 45º
Rotating nose piece
• Fore lenses Attached to a rotating nose piece, at the base of the body tube are a group of 3
or 4 in some microscopes 5 objectives.
• Locate the disc and notice a click as each objective snap into position.
• The objective lenses focus the light that comes through the specimen, up the body tube,
and through the oculars.
• Each objective has numbers stamped on it.
• One of these numbers identifies the magnification of the objective (e.g., 40X).
• Objective lenses are usually named according to their magnifying power,
• as follows: the red , scanning power 4X; the yellow low power 10X; The blue high power
43X; the wight , oil immersion 100X
• A 2nd number on the objective, usually a decimal represents the numerical aperture for
that lens; the abbreviation NA may precede the number.
• The total magnification for each objective is calculated by multiplying the magnification
of the ocular and objective lens on your microscope.
• OCULAR M.P. X OBJECTIVE M.P. = TOTAL MAGNIFICATION PWER
Rotating nose piece
• Objective lenses are usually named according to their magnifying power,
• as follows: the red , scanning power 4X; the yellow low power 10X; The
blue high power 43X; the wight , oil immersion 100X
• A 2nd number on the objective, usually a decimal represents the numerical
aperture for that lens; the abbreviation NA may precede the number.
• The total magnification for each objective is calculated by multiplying the
magnification of the ocular and objective lens on your microscope.
• OCULAR M.P. X OBJECTIVE M.P. = TOTAL MAGNIFICATION PWER
Rotating nose piece
Mechanical Stage:
• The surface or platform on which you place the microscope slide is the
stage.
• Note the opening in the center of the stage.
• the stage is movable and is called a mechanical stage.
• Movement is controlled by 2 knobs located on the top side or bottom of
the stage.
• Note the horizontal and vertical scales on the mechanical stage.
• The mechanical stage on a microscope moves in three dimensions.
First, it moves up and down on the vertical axis to obtain focus.
Mechanical Stage:
It also moves on the X-Y
axis((Right, left, forward and
backward)), to allow you to
‘scan’ your specimen at the
horizontal axis . you can
make fine movements of the
slide to achieve optimal
viewing positioning for your
experiments.
Substage:
• The area under the stage, called the substage, contain a
diaphragm, a condenser, or both.
• Diaphragm: The diaphragm regulates the amount of light
passing from the light source through the specimen and
through the lens system of the microscope.
• By properly adjusting the diaphragm, you can provide
better contrast between the surrounding medium and your
specimen, thus greatly improving your image of the
specimen.
• An iris diaphragm consists of a circle of overlapping thin
metal plates. The lever that projects from the side of the
diaphragm opens and closes these plates, thereby regulating
the amount of light that enters the microscope.
Condenser
• The condenser consists of a series of lenses that
focus light onto the specimen.
• It is moved up and down by a knob at its side or
by a lever projecting from the condenser
housing.
• By properly adjusting the condenser, you can
greatly improve the clarity of the specimen.
• They act to gather light from the microscope's
light source and concentrate it into a cone of
light that
illuminates the specimen
A filter holder may be attached to the bottom of the condenser.
It usually contains a blue filter.
The base :
A sold flat metal part support the microscope
contain
the light source ,
brightness control.
on\off switch the and
the electric cable open the
arm held above base and holding all other part
of microscope .
The Light Source:
• The microscope has an illuminator held
into the base of the microscope and
controlled by an on/off switch.
• You can control the amount of light
entering the specimen by adjusting the
diaphragm.
• You can also control the light intensity
by adjusting the voltage of a transformer
or ( brightness adjustment) attached to the
illuminator.
• Use low or medium voltage settings for
most microscopic observations.
• You will need a higher setting when using
the oil-immersion lens.
The coarse and fine adjustment knobs.
• Coarse Adjustment Knob- The coarse adjustment
knob located on the arm of the microscope moves the
stage up and down to bring the specimen into focus.
• The gearing mechanism of the adjustment produces a
large vertical movement of the stage with only a partial
revolution of the knob.
• Because of this, the coarse adjustment should only be
used with low power (4X and 10X objectives) and
never with the high power lenses (40X and 100X).
• Fine Adjustment Knob - This knob is inside the
coarse adjustment knob and is used to bring the
specimen into sharp focus under low power and is
used for all focusing when using high power lenses
The high power oil immersion lens :
• when using the high power 100X lens in microscope the image do not
appear clearly because of the Light travels at different speeds in air and in
glass (faster in air which is usually taken as the standard of 1).
• Oil has a refractive index of 1.515 like common glass refractive index .
• so the air causing a refraction in light that travel from the slide(glass) or
( liquid sample ) to the air towards to the objective lens 100X .
• this deflection or refraction causing the specimens image is not appear
clearly .
• oil with refraction index like common the glass refractive index when full
the space between sample and the objective lens keep light rays in straight
line in this way will see clear image from the specimen under 100x
magnifying in the microscope .
The high power oil immersion lens :
