St.

Paul University Surigao

St. Paul University System
8400 Surigao City, Philippines

Name: __________________________________ Date: _____________________________

Course & Year: ___________________________ Professor: _________________________

MICROSCOPY
Activity No. 1

I. Learning Outcomes:
At the end of the activity the students should be able to:
1. describe the different types of microscopes in terms of their uses;
2. name and give the functions of the basic parts of the compound light microscope; and
3. calculate the total magnification for both low- and high-power lens systems.

II. Microscopy

Because biological objects can be very small, we often use a microscope to view
them. Many kinds of instruments, ranging from the hand lens to the electron microscope,
are effective
magnifying devices.

Light Microscopes

Light microscopes use visible light rays that are magnified and focused by means
of lenses. The binocular dissecting microscope (stereomicroscope) is designed to
study entire objects in three dimensions at low magnification. The compound light
microscope is used for examining small or thinly sliced sections of objects under higher
magnification than that of the binocular dissecting microscope. The term compound
refers to the use of two sets of lenses: the ocular lenses located near the eyes and the
objective lenses located near the object. Illumination is from below, and visible light passes
through clear portions but does not pass-through opaque portions. To improve contrast,
the microscopist uses stains or dyes that bind to cellular structures and absorb light. Figure
2.5a is a photomicrograph, a photograph of an image produced by a compound light
microscope.

Figure 1.1 Comparative micrographs

Micrograph of a lymphocyte, a type of white blood cell. The photomicrograph in (a)

shows a less detail than the transmission electron micrograph (TEM) in (b). The scanning
electron micrograph (SEM) (c) shows the cell surface in three dimensions.

a. photomicrograph b. transmission electron micrograph c. scanning electron micrograph

 St. Paul University Surigao
St. Paul University System
8400 Surigao City, Philippines

Rules for Microscope Use

Observe the following rules for using a microscope:
1. The lowest power objective (scanning or low) should be in position both at the
beginning and
at the end of microscope use.
2. Use only lens paper for cleaning lenses.
3. Do not tilt the microscope as the eyepieces could fall out, or wet mounts could be
ruined.
4. Keep the stage clean and dry to prevent rust and corrosion.
5. Do not remove parts of the microscope.
6. Keep the microscope dust-free by covering it after use.
7. Report any malfunctions.

Use of the Compound Light Microscope

As mentioned, the name compound light microscope indicates that it uses two sets
of lenses and light to view an object. The two sets of lenses are the ocular lenses located
near the eyes and the objective lenses located near the object. Illumination is from below,
and the light passes through clear portions but does not pass-through opaque portions.
This microscope is used to examine small or thinly sliced sections of objects under higher
magnifications than would be possible with the binocular dissecting microscope.

Identifying the Parts

Identify the following parts on your microscope and label them in Figure 2.6. Label this
microscope with the help of the text material.

Figure 1.2 Standard Microscope

 St. Paul University Surigao
St. Paul University System
8400 Surigao City, Philippines

1. Eyepiece (ocular lens): Usually the magnifying power of the ocular lens of the
microscope is 10x.
2. Body tube: Holds nosepiece at one end and eyepiece at the other end; conducts
light rays.
3. Arm: Supports upper parts and provides carrying handle.
4. Nosepiece: Revolving device that holds objectives.
5. Objectives (objective lenses):

a. Scanning power objective: This is the shortest of the objective lenses and is
used to scan the whole slide. The magnification is stamped on the housing of the
lens. It is a number followed by an x.

b. Low-power objective: This lens is longer than the scanning lens and is used to
view objects in greater detail.

c. High-power objective: If your microscope has three objective lenses, this lens
will be the longest. It is used to view an object in even greater detail.

d. Oil immersion objective (on microscopes with four objective lenses). Holds a
95x (to 100x) lens and is used in conjunction with immersion oil to view objects
with the greatest magnification.

4X – This objective magnifies the image by a factor of 4. It is referred to as

the “scanning objective” since it is used to scan the slide to locate
the specimen before viewing it at higher magnification. Your
microscope may not have this objective lens, in which case you
can begin with the 10X objective.
10X – This objective magnifies the image by a factor of 10 and is referred
to as the “low power” objective.
43X (or 45X) – This objective magnifies the image by a factor of 43 (or 45)
and is referred to as the “high power” objective.
100X – This objective magnifies the image by a factor of 100. It is referred
to as the “oil immersion objective” since it requires a drop of
immersion oil on the slide to provide good resolution. You will not
be using this objective lens.

6. Coarse-adjustment knob: Knob used to bring object into approximate focus;

used only with low-power objective.
7. Fine-adjustment knob: Knob used to bring object into final focus.
8. Condenser: Lens system below the stage used to focus the beam of light on
the object being viewed.
9. Diaphragm of diaphragm control lever: Controls amount of illumination used
to view the object.
10. Light Source: An attached lamp that directs a beam of light up through the
object.
11. Base: The flat surface of the microscope that rests on the table.
12. Stage: Holds and supports microscope slides.
13. Stage clips: Holds slides in place on the stage.
14. Mechanical stage (optional): movable stage that aids in the accurate
positioning of the slide.
15. Mechanical stage control knobs (optional): Two knobs that are usually
located below the stage. One knob control forward/reverse movement, and the
other controls right/left movement.
 St. Paul University Surigao
St. Paul University System
8400 Surigao City, Philippines

How do you calculate magnification on a light microscope?

To calculate the total magnification of the compound light microscope, multiply

the magnification power of the ocular lens by the power of the objective lens. For
instance, a 10x ocular and a 40x objective would have a 400x total magnification. The
highest total magnification for a compound light microscope is 1000x

Calculating Magnification

Once the magnification of each individual lens is known, calculating total

magnification is simple math. Multiply the magnification of the lenses together. For
example, if the eyepiece magnification is 10x and the objective lens in use has a
magnification of 4x, the total magnification is:
10 times 4 = 40 10×4=40

The total magnification of 40 means that the object appears forty times larger than
the actual object. If the viewer changes to the 10x objective lens, the total magnification
will be the ocular's 10x magnification multiplied by the new objective lens's 10x
magnification, calculated as:
10 times 10 = 100 10×10=100

III. Problem Solving: Show Solution

1. Calculate the highest total magnification for a microscope with an ocular lens of 5X
and objective lenses of 5X, 10X, and 30X.

2. Calculate the highest total magnification for a microscope with an ocular lens of 10X
and objective lenses of 15 X, 4 X, and 20X.

IV. Conclusion