Republic of the Philippines

Department of Education
Region 02- Cagayan Valley
Schools Division of the City of Ilagan
ISABELA NATIONAL HIGH SCHOOL

LEARNING ACTIVITY SHEET

BIOTECHNOLOGY GRADE 8- STE
LESSON 6: MICROSCOPE
Week 8

Name:_____________________________________________ Date:________________________

Grade Level & Section:________________________________ Score:_______________________

INTRODUCTION:

Microscopes are instruments designed to produce magnified visual or photographic images of

objects too small to be seen with the naked eye. The microscope must accomplish three tasks: produce a
magnified image of the specimen, separate the details in the image, and render the details visible to the
human eye or camera. This group of instruments includes not only multiple-lens (compound microscopes)
designs with objectives and condensers, but also very simple single lens instruments that are often hand-
held, such as a loupe or magnifying glass.

The microscope illustrated in Figure 1 is a simple compound microscope invented by British microscopist
Robert Hooke sometime in the 1660s. This beautifully crafted microscope has an objective lens near the
specimen and is focused by turning the body of the microscope to move the objective closer to or farther
from the specimen. An eyepiece lens is inserted at the top of the microscope and, in many cases, there is
an internal "field lens" within the barrel to increase the size of the viewfield. The microscope in Figure 1
is illuminated through the oil lamp and water-filled spherical reservoir, also illustrated in Figure 1. Light
from the lamp is diffused when it passes through the reservoir and is then focused onto the specimen
with a lens attached to the reservoir. This early microscope suffered from chromatic (and spherical)
aberration, and all images viewed in white light contained "halos" that were either blue or red in color.

GENERAL OBJECTIVES/ LEARNING COMPETENCIES:

❖ Differentiate the types of microscope and it’s uses.

LESSON PROPER:

Structural parts of a microscope and their functions

Figure: Diagram of parts of a microscope

There are three structural parts of the microscope i.e. head, base, and arm.
1. Head – This is also known as the body, it carries the optical parts in the upper part of the
microscope.
2. Base – It acts as microscopes support. It also carriers the microscopic illuminators.
3. Arms – This is the part connecting the base and to the head and the eyepiece tube to the base of
the microscope. It gives support to the head of the microscope and it also used when carrying the
microscope. Some high-quality microscopes have an articulated arm with more than one joint
allowing more movement of the microscopic head for better viewing.
Optical parts of a microscope and their functions
The optical parts of the microscope are used to view, magnify, and produce an image from a specimen
placed on a slide. These parts include:
1. Eyepiece – also known as the ocular. this is the part used to look through the microscope. It is
found at the top of the microscope. Its standard magnification is 10x with an optional eyepiece
having magnifications from 5X – 30X.
2. Eyepiece tube – it is the eyepiece holder. It carries the eyepiece just above the objective lens. In
some microscopes such as the binoculars, the eyepiece tube is flexible and can be rotated for
maximum visualization, for variance in distance. For monocular microscopes, they are not flexible.
3. Objective lenses – These are the major lenses used for specimen visualization. They have a
magnification power of 40x-100X. There are about 1- 4 objective lenses placed on one
microscope, in that some are rare facing and others face forward. Each lens has its own
magnification power.
4. Nose piece – also known as the revolving turret. It holds the objective lenses. It is movable hence
it can revolve the objective lenses depending on the magnification power of the lens.
 5. The Adjustment knobs – These are knobs that are used to focus the microscope. There are two
types of adjustment knobs i.e fine adjustment knobs and the coarse adjustment knobs.
6. Stage – This is the section on which the specimen is placed for viewing. They have stage clips hold
the specimen slides in place. The most common stage is a mechanical stage, which allows the
control of the slides by moving the slides using the mechanical knobs on the stage instead of
moving it manually.
7. Aperture – This is a hole on the microscope stage, through which the transmitted light from the
source reaches the stage.
8. Microscopic illuminator – This is the microscopes light source, located at the base. It is used
instead of a mirror. it captures light from an external source of a low voltage of about 100v.
9. Condenser – These are lenses that are used to collect and focus light from the illuminator into the
specimen. They are found under the stage next to the diaphragm of the microscope. They play a
major role in ensuring clear sharp images are produced with a high magnification of 400X and
above. The higher the magnification of the condenser, the more the image clarity. More
sophisticated microscopes come with an Abbe condenser that has a high magnification of about
1000X.
10. Diaphragm – it is also known as the iris. It is found under the stage of the microscope and its
primary role is to control the amount of light that reaches the specimen. It is an adjustable
apparatus, hence controlling the light intensity and the size of the beam of light that gets to the
specimen. For high-quality microscopes, the diaphragm comes attached with an Abbe condenser
and combined they are able to control the light focus and light intensity that reaches the
specimen.
11. Condenser focus knob – this is a knob that moves the condenser up or down thus controlling the
focus of light on the specimen.
12. Abbe Condenser – this is a condenser specially designed on high-quality microscopes, which
makes the condenser to be movable and allows very high magnification of above 400X. The high-
quality microscopes normally have a high numerical aperture than that of the objective lenses.
13. The rack stop – It controls how far the stages should go preventing the objective lens from getting
too close to the specimen slide which may damage the specimen. It is responsible for preventing
the specimen slide from coming too far up and hit the objective lens.
Types of Microscopes
A good microscope should have three properties:
1. Good resolution: Resolution power refers to the ability to produce separate images of closely
placed objects so that they can be distinguished as two separate entities. The resolution power
of:
• The unaided human eye is about 0.2 mm (200 μm)
• The light microscope is about 0.2 μm
• An electron microscope is about 0.5 nm
The resolution depends on refractive index. Oil has a higher refractive index than air.
2. Good contrast: This can be further improved by staining the specimen.
3. Good magnification: This is achieved by the use of concave lenses.
Bright-Field or Light Microscope
The bright-field or light microscope forms a dark image against a brighter background.
Dark Field Microscope
Principle: In a dark field microscope, the object appears bright against a dark background. This is made
possible by the use of a special darkfield condenser.
Applications: It is used to identify the living, unstained cells and thin bacteria like spirochetes which
cannot be visualized by light microscopy.
Phase Contrast Microscope
It is used to visualize the living cells by creating a difference in contrast between the cells and water. It
converts slight differences in refractive index and cell density into easily detectable variations in light
intensity.
It is useful for studying:
• Microbial motility
• Determining the shape of living cells
• Detecting bacterial components such as endospores and inclusion bodies.
Fluorescence Microscope
Principle: When fluorescent dyes are exposed to ultraviolet rays (UV) rays, they become excited and are
said to fluoresce, i.e. they convert this invisible, short-wavelength rays into the light of longer
wavelengths (visible light).
Applications: Epifluorescence microscope has the following applications:
• Autofluorescence, when placed under UV lamp, e.g. Cyclospora
• Microbes coated with the fluorescent dye, e.g. Acridine orange for malaria parasites (QBC) and
Auramine phenol for M. tuberculosis.
• Immunofluorescence: It uses a fluorescent dye tagged antibody to detect cell surface antigens
or antibodies bound to cell surface antigens. There are three types: direct IF, indirect IF, and
Flow cytometry.
Electron Microscope
It was invented by Ernst Ruska in 1931. It differs from a light microscope in various ways.
There are two types of EM:
• Transmission EM (MC type, examine the internal structure, resolution 0.5 nm, gives 2-
dimensional view)
• Scanning EM (examine the surfaces, resolution 7 nm, gives 3-dimensional view)
Principle of Transmission Electron Microscope
Specimen preparation: Cells are subjected to the following steps to prepare very thin specimens (20 to
100 nm thick)
• Fixation: Cells are fixed by using glutaraldehyde or osmium tetroxide for stabilization.
• Dehydration: Specimen is then dehydrated with organic solvents (e.g. acetone or ethanol).
• Embedding: Specimen is embedded in plastic polymer and then, is hardened to form a solid
block. Most plastic polymers are water-insoluble; hence complete dehydration of specimen is a
must before embedding.
• Slicing: Specimen is then cut into thin slices by ultramicrotome knife, and slices are mounted on
a metal slide (copper).
Freeze-etching technique: It is an alternate method for specimen preparation to visualize the internal
organelles within the cells.
Cells are rapidly frozen then warmed → fractured by a knife exposing the internal organelles →
subjected to sublimation → shadowed by coating with platinum and carbon.
Measures to increase the contrast of EM include:
• Staining by solutions of heavy metal salts like lead citrate and uranyl acetate
• Negative staining with heavy metals like phosphotungstic acid or uranyl acetate.
• Shadowing: Specimen is coated with a thin film of platinum or other heavy metal at a 45° angle
so that the metal strikes the microorganism on only one side.

ACTIVITY 11

Give what is being asked in each question:

1. Define a Microscope.
2. State functions of a microscope.
3. Diagrammatically, identify the various parts of a microscope.
4. Describe the functions of each part of the microscope you have drawn above.
5. Differentiate between a condenser and an Abbe condenser.
6. What is the magnification power of the objective lenses?
7. How does the eyepiece compare to the objective lens?
8. Why is the rack stop included in the microscope from the factory and can it be replaced?
9. Differentiate the types of microscope.
10. What is the most common type of microscope being used today?
 Activity 12
Microscope Labeling

Activity 13:
Microscope Use:

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. You should carry the microscope by the ____________ and the ______________.
5. The objectives are attached to what part of the microscope (it can be rotated to click lenses into
place?) ____________________________________________
6. A microscope has an ocular objective of 10x and a high power objective of 50x, what is the
microscope's total magnification? ______________

References: biorender.com

Prepared by:

CRISTINA D. ANGANGAN
Teacher II

NOTE: Practice Personal Hygiene Protocol at all times.