The Microscope

Dr. Riaz Hussain Pasha

 The History
• The first microscope was 6 feet long!!! (1590)

• The Greeks & Romans used “lenses” to

magnify objects over 1000 years ago.

• Hans and Zacharias Janssen of Holland in

the 1590’s created the “first” compound
microscope
 The History
• Anthony van Leeuwenhoek and Robert
Hooke made improvements by working
on the lenses

Anthony van Leeuwenhoek Hooke Microscope Robert Hooke

1632-1723 1635-1703
 The History

Zacharias Jansen The “First” Microscope

1588-1631
 How a Microscope Works
Convex Lenses are
curved glass used to
make microscopes
(and glasses etc.)

Convex Lenses bend

light and focus it in
one spot.
 How a Microscope Works
Ocular Lens Objective Lens
(Magnifies Image) (Gathers Light,
Magnifies
And Focuses Image
Body Tube Inside Body Tube)
(Image Focuses)

•Bending Light: The objective (bottom) convex lens

magnifies and focuses (bends) the image inside the
body tube and the ocular convex (top) lens of a
microscope magnifies it (again).
The Parts of a Microscope
 Ocular Lens

Body Tube

Nose Piece
Arm
Objective
Lenses
Stage
Stage
Clips
Coarse Adj.

Diaphragm Fine Adjustment

Light Source
Base

Skip to Magnification Section

 Body Tube
• The body tube holds the objective
lenses and the ocular lens at the proper
distance

Diagram
 Nose Piece
• The Nose Piece holds the objective
lenses and can be turned to increase
the magnification

Diagram
 Objective Lenses
• The Objective Lenses increase
magnification (usually from 4x,10x,40x,
100x)

Diagram
 Stage Clips
• These 2 clips hold the slide/specimen in
place on the stage.

Diagram
 Diaphragm
• The Diaphragm controls the amount of
light on the slide/specimen

Turn to let more light in or to

make dimmer.

Diagram
 Light Source
• Projects light upwards through the
diaphragm, the specimen and the
lenses
• Some have lights, others have mirrors
where you must move the mirror to
reflect light

Diagram
 Ocular Lens/Eyepiece
• Magnifies the specimen image

Diagram
 Arm
• Used to support the microscope when
carried. Holds the body tube, nose
piece and objective lenses

Diagram
 Stage
• Supports the slide/specimen

Diagram
 Coarse Adjustment Knob/screw
• Moves the stage up and down (quickly)
for focusing your image

Diagram
 Fine Adjustment Knob
• This knob moves the stage SLIGHTLY
to sharpen the image

Diagram
 Base
• Supports the microscope

Diagram
 Resolving Power
• It is a measure of the capacity of a microscope
to separate two objects lying close to each
other.

• R=0.61λ/NA

• Where: R= Resolving power, λ=Wave length,

NA= Numerical Aperture of the objective lens
 Magnification
• It is the ratio of image size to object size
in terms of linear distance
• To determine your magnification…you
just multiply the power of ocular lens by
the objective lens
• Ocular 10x Objective 40x:10 x 40 =
400 So the object is 400 times “larger”
Objective Lens have
their magnification
written on them.

Ocular lenses usually magnifies by 10x

 Comparing Powers of Magnification

We can see better details with higher the powers of

magnification, but we cannot see as much of the image.

Which of these images would

be viewed at a higher power
of magnification?
 Caring for a Microscope
• Clean only with a soft cloth/tissue

• Make sure it’s on a flat surface

• Carry it with 2 HANDS…one on the arm

and the other on the base
 Carry a Microscope Correctly

Always carry a microscope with one hand holding

the arm and one hand under the base.
 Use of Microscope ...
• 1 – Turn on the microscope and then rotate the
nosepiece to click the objective into place.
• 2 – Inspect slide with naked eye, handle slide by
corners, place it on the stage and secure it using the
stage clips. Lower scan to its minimum. Use the coarse
adjustment knob (large knob) to get it the image into
view and then use the fine adjustment knob (small knob)
to make it clearer.
• 3 – Once you have the image in view, rotate the
nosepiece to view it under different powers.
• 4 – When you are done, turn off the microscope and put
up the slides you used.
 Precautions…..
• Start on the lowest magnification
• Don’t use the coarse adjustment knob on high
magnification…you’ll break the slide!!!
• Place slide on stage and lock clips
• Clean the microscope before and after its use
• Avoid excessive rubbing of lens
• Keep the microscope covered
• Lower the lens down to the object, then move the
lens upwards and focus
 Staining
• A technique used in microscopy to
enhance the contrast in the microscopic
image
• Stains and dyes (coloring substance)
are frequently used in biology and
medicine to highlight structures in
biological tissues for viewing, often with
the aid of different microscopes
• Basically two types; IN-VIVO (vital
staining) & IN-VITRO techniques
• Acidic dye: dye which is having staining
property in acid radicals e.g., Eosin,
picric acid, Orange G, Trypan blue…
etc
• Basic dye: which is having reactive
basic radical (having coloring property)
e.g., Hematoxylin, Toluidine blue,
methylene blue… etc
• Amphoteric dye: containing both
reactive acidic and basic radicals and
stain both acidic & basic elements e.g.,
Celestine blue B….
• Metachromatic dye: stains the tissues
two or more color e.g., Azure A,
Thionin, methylene blue … etc
• Basophilic: The structures which stain
with basic dye
• Acidophilic: The tissue or components
of cell which stain with acid dye
• Argyrophilic: structures stained by silver
stain e.g., reticular fibers