What is a microscope?
OBJECTIVES
• Explain the principle of microscopy
• Identify and state the functions of a compound
microscope
• Identify the different microscopes and their
functions
• Demonstrate and discuss the proper care and use
of a microscope
The microscope is
probably one of the pieces
of equipment that is mostly
used in a clinical laboratory
In simple terms – a microscope is a
magnifying glass. Enlarge very small
objects.
Compound light
Microscope/Brightfield microscope
What is a microscope?
• Light Microscope - uses
compound (2) lenses to
magnify objects. The
lenses bend or refract
light to make the object
beneath them appear
closer.
Micro- = “small”; -scope = “to look
at”
Types of Light Microscopes
1.Bright field Light Microscope
2.Phase Contrast Light Microscope
3.Dark-Field Light Microscope
4.Fluorescence Light Microscope
Types of Light Microscopes
1.Bright field Light Microscope
2.Phase Contrast Light Microscope
3.Dark-Field Light Microscope
4.Fluorescence Light Microscope
Typical Classroom Microscope
PARTS OF A MICROSCOPE
1.Support System
2.Magnification System
3.Illumination System
4.Adjustment System
Support System
• The foot/Base
• The arm
• The revolving nosepiece
• The mechanical stage
• The stage clip
Base
Base
• Supports the whole microscope
• Used to carry the microscope
• When carrying a microscope, always
have one hand on the arm and one
hand on the base. Use two hands!!
Arm
Arm
• Supports the upper parts of the
microscope
• Used to carry the microscope
• When carrying a microscope, always
have one hand on the arm and one
hand on the base. Use two hands!!
Nosepiece
Nosepiece
• Holds the objective lenses
• Rotates to enable magnification
• Located at the bottom of the body tube
Stage
Stage
• Supports the slide
• The slide contains the specimen or
object that you are viewing with the
microscope.
Stage Clip
Stage Clip
• Helps to hold the slide in place
• Usually one on each side of the hole
(stage opening) = 2 stage clips
• The stage opening allows light to pass
from the light source to the lenses.
MAGNIFICATION SYSTEM
• Eyepiece
• Objectives
Eyepiece
Eyepiece
• Also known as the ocular
• Contains the first lens you look through
- usually a magnification of 10x
• Located on the top of the body tube
THE EYEPIECE
(a)Magnification
The magnifying power of the eyepiece is marked on it:
x 4 eyepiece magnifies the image 4 times
x 6 eyepiece magnifies the image 6 times
x 10 eyepiece magnifies the image 10 times
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Objective Lenses
Objective Lenses
• Used in combination with the eyepiece
to provide a range of magnification
• Magnification ranges from 4x to 100x
• Located on the nose-piece at the
bottom of the body tube
• Colour coded
What is magnification?
Magnification is a measure of how much larger a microscope (or set of lenses
within a microscope) causes an object to appear.
Magnification is defined by the
magnification by the objective
x
the magnification by eyepiece
BUT maximum magnification does not mean maximum resolution!
What is resolution?
Numerical aperture, NOT magnification determines resolution!
A lens with a larger NA will be able to visualize finer details and
will also collect more light and give a brighter image than a lens
with lower NA.
OBJECTIVES
(b)The numerical aperture (NA)
The numerical aperture is also engraved on the sleeve,
next to the magnification eg.
0.30 on the x 10 objective
0.65 on the x 40 objective
1.30 on the x 100 objective
The greater the numeric aperture, the greater the resolving
power (the ability to reveal closely adjacent details as
separate and distinct). 29
Magnification
Your microscope has 3 magnifications (objective lenses):
Scanning
Low
High
Each objective has its magnification labeled. In addition to this, the
_______ Lens (eyepiece) has a magnification of __X.
Ocular ______ 10
To calculate total magnification, ________
multiply the ocular x objective.
• Resolving power
• The greater the resolving power of the objective, the
clearer the image and the greater the ability to reveal
closely adjacent or nearby details as separate and clear.
• The maximum resolving power of a good medical
laboratory microscope is about 0,25 um.
• (the resolving power of the normal human eye is 0,25
mm).
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Illumination system
• Source of light – bulb
• Condenser – brings the ray of lights to a common focus
on the object to be examined
• Iris Diaphragm – reduces or increases the angle
therefore the amount of light that passes into the
condenser
• Filters – some microscopes have coloured filters below
the condenser
Light Source
Light Source
• Provides light necessary for viewing
the specimen
• Sends light through the stage opening
to the diaphragm
Diaphragm
Diaphragm
• Wheel or lever located below the stage
opening
• Regulates the amount of light that can
enter the lenses
• May need to be adjusted based on the
thickness of the specimen being
studied
Adjustment System
• Course adjustment knob
• Fine adjustment knob
Coarse Adjustment
Knob
Coarse Adjustment Knob
• Raises and lowers the stage or
objective lenses
• Used only when focusing the low
power (4x) objective lens
Fine Adjustment Knob
Fine Adjustment Knob
• Raises and lowers the stage or
objective lenses a small distance for
exact focusing
• Used when focusing the medium
power (10x) and high power (40x)
objective lenses
Recap
1. Scanning --> use coarse and fine knob DO NOT SKIP
2. Low power --> use coarse and fine knob STEPS!!!!
3. High power --> use fine knob only
What will happen if I use the course knob
under high
The power?
slide or lens could crack
ROUTINE MAINTENANCE OF
THE MICROSCOPE
• Microscope needs DAILY attention!!
• Items needed:
Tissue paper
Special lens tissue
Alcohol
Plastic cover
Let’s Review...
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Quiz Over the Microscope
1. When focusing a specimen, you should always start with the
___________________ objective.
2. When using the high power objective, only the ________
___________ knob should be used.
3. The type of microscope used in most science classes is the
_________________ microscope
4. What part of the microscope can adjust the amount of light
that hits the slide? ______________________________
5. You should carry the microscope by the ________ and the
__________.
6. The objectives are attached to what part of the microscope (it
can be rotated to click the lenses into place):
_______________ ________________
7. You should always store your microscope with the
________________ objective in place.
8. A microscope has an ocular objective of 10x and a high
power objective of 50x. What is this microscope's total
magnification? ____________
SELF DIRECTED STUDY
• Phase contrast microscopy
• Fluorescent microscopy
• Electron Microscopy
• Dark Field Microscopy
