PRINCIPLES OF

MICROSCOPY
BY: IVAN ABINAL
 INTRODUCTION

 A compound microscope has multiple lenses: the objective lens (typically 4x, 10x, 40x or 100x) is
compounded (multiplied) by the eyepiece lens (typically 10x) to obtain a high magnification of 40x,
100x, 400x and 1000x. Higher magnification is achieved by using two lenses rather than just a single
magnifying lens.
 As well as being the father of microbiology, van Leeuwenhoek laid the foundations of plant anatomy
and became an expert on animal reproduction. He discovered blood cells and microscopic
nematodes, and studied the structure of wood and crystals. He also made over 500 microscopes to
view specific objects.
 LEARNING OUTCOMES

 Explain how do we take care of our microscopes and what are the ways to store our
microscope.
 How to take care of microscopes

• Do not touch the glass part of the lenses with your fingers. Use only special lens paper to clean
the lenses.
• Always keep your microscope covered when not in use.
• Always carry a microscope with both hands. Grasp the arm with one hand and place the other
hand under the base for support.
 How to store microscopes

1.Put the low power objective into place.

2.Lower the stage.
3.Take the slide off the stage.
4.Turn the microscope off.
5.Wrap the cord.
6.Cover and push the microscope to the back of the lab bench (to a safe location)
7.Clean lab station.
 SPECIFIC OUTCOME

 Explain the function of the parts of compound microscope.

• Eyepiece – also known as the ocular. ...

• Eyepiece tube – it's the eyepiece holder. ...
• Objective lenses – These are the major lenses used for specimen visualization. ...
• Nose piece – also known as the revolving turret. ...
• The Adjustment knobs – These are knobs that are used to focus the microscope.
•Specimen or slide: The object used to hold the specimen in place along with slide covers for viewing. Most slides & slide
covers are thin glass rectangles.
•Stage or Platform: The platform upon which the specimen or slide are placed. The height of the mechanical stage is
adjustable on most compound microscopes.
•Stage clips or mechanical stage: Clips on the stage that hold the slide in place on the mechanical stage.
•Aperture - Disc or Iris Diaphragm: Circular opening in the stage where the illumination from the base of the compound
microscope reaches the platform of the stage.
•Abbe Condenser: This lens condenses the light from the base illumination and focuses it onto the stage. This piece of the
compound microscope sits below the stage & typically acts as a structural support that connects the stage to arm or frame of the
microscope.
•Coarse and fine adjustment controls: Adjusts the focus of the microscope. These knobs increase or decrease the level of
detail seen when looking at the slide or specimen through the eyepiece of the compound microscope.
•Stage height adjustment: Adjusts the position of the mechanical stage vertically & horizontally. It is important to adjust these
knobs so that the objective lens is never coming into contact with the slide or specimen on the stage.
•Mirror: Reflects light into the base of the microscope. Earlier microscopes used mirrors that reflected light into the base of
the microscope instead of halogen bulbs as their source of illumination.
•Illumination: Light used to illuminate the slide or specimen from the base of the microscope. Low voltage halogen bulbs are
the most commonly used source of illumination for compound microscopes.
•Bottom Lens or Field Diaphragm: Knob used to adjust the amount of light that reaches the specimen or slide from the base
illumination.
 Define the following

 Magnification power
 Resolving power
 Light intensity
 Field of view
 Dept of field focus
 Working distance
 Magnifying power

Magnifying power is defined as the ratio between the dimensions of the image and the
object. The process of magnification can occur in lenses, telescopes, microscopes and even
in slide projectors. Simple magnifying lenses are biconvex - these lenses are thicker at the
center than at the edges
 Resolving power

 Resolving power denotes the smallest detail that a microscope can resolve when
imaging a specimen; it is a function of the design of the instrument and the properties of
the light used in image formation. ... The smaller the distance between the two points that
can be distinguished, the higher the resolving power.
 Light intensity

 External Websites. Luminous intensity, the quantity of visible light that is emitted in

unit time per unit solid angle. The unit for the quantity of light flowing from a source in
any one second (the luminous power, or luminous flux) is called the lumen. The lumen is
evaluated with reference to visual sensation.
 Field of view

 Field of view(FOV) is the maximum area of a sample that a camera can image. It is
related to two things, the focal length of the lens and the sensor size.The focal length of the
lens describes the distance between the lens and the focused image on the sensor(FOV)
is the maximum area of a sample that a camera can image. It is related to two things,
the focal length of the lens and the sensor size. The focal length of the lens describes the
distance between the lens and the focused image on the sensor
 Depth of field

 For many cameras, depth of field (DOF) is the distance between the nearest and the farthest
objects that are in acceptably sharp focus in an image. The depth of field can be calculated
based on focal length, distance to subject, the acceptable circle of confusion size, and aperture.
 Working distance

 Working distance is the distance between the front of the microscope objective lens
and the surface of the specimen or slide coverslip at the point where the specimen is
completely in focus.
 Determine the specific techiques in using, cleaning, and
storing compound microscopes in the laboratory

By applying the tips of how to take care microscopes and at the same time on how to
proper storing it.
 Guide questions

 1. Determine the function.

2.What knobs do you see to move the stage up and down when you see following objective?
- letter B
3.Give the magnification power of this three lenses.
a. ocular lens/eyepiece (100x)
b.LPO (100X)
c.HPO (400X)
d.scanning objectives (40x)
How do you control the light that enters in the viewing area?

  Iris Diaphragm controls the amount of light reaching the specimen. It is located

above the condenser and below the stage. Most high quality microscopes include an
Abbe condenser with an iris diaphragm.
II. VIEWING OF SPICEMEN IN THE MICROSCOPE

 DEFINE THE FOLLOWING PRINCIPLES OF THE MICROSCOPE

As I said earlier, there are56 principle of microscopes and there no need to
explain 1 by 1 because I already tackled that one.
 Give the do’s and don’ts in using compound microscope

 DO’S

ways carry the microscope with two hands. One goes under the base and the other holds the arm.

Place the microscope away from the edge of the table with the arm facing toward you.

ONLY use lens paper to clean the lenses of the microscope.

DON’T’S

1.Poor slide preparation. A smudged slide.

2.Insufficient magnification. Insufficient magnification.
3.Stay focussed. The last common issue is the out-of-focus image.
4.Centre the lifeforms in the middle of your slide.