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Microscopy Lecture 2024

A microscope is a magnifying instrument used to enlarge small objects, commonly utilized in clinical laboratories. It consists of various systems including support, magnification, illumination, and adjustment systems, with different types such as bright field, phase contrast, and fluorescence microscopes. Proper care and usage are essential, including routine maintenance and understanding the functions of its components for effective operation.

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Angelina Human
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Microscopy Lecture 2024

A microscope is a magnifying instrument used to enlarge small objects, commonly utilized in clinical laboratories. It consists of various systems including support, magnification, illumination, and adjustment systems, with different types such as bright field, phase contrast, and fluorescence microscopes. Proper care and usage are essential, including routine maintenance and understanding the functions of its components for effective operation.

Uploaded by

Angelina Human
Copyright
© © All Rights Reserved
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
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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

24
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).

31
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...
1

2 4
3
5
12
6
7 8
9 10

11
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

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