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Bio101 Lab Activity 03 - The Microscope

The document describes how to identify the parts and functions of a compound light microscope. It provides instructions on proper use of the microscope, including focusing, adjusting lenses, and determining total magnification and field of view size. Students will learn to identify microscope parts, describe proper microscope care and use, calculate total magnification using ocular and objective lenses, and determine field of view diameter and area.

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164 views

Bio101 Lab Activity 03 - The Microscope

The document describes how to identify the parts and functions of a compound light microscope. It provides instructions on proper use of the microscope, including focusing, adjusting lenses, and determining total magnification and field of view size. Students will learn to identify microscope parts, describe proper microscope care and use, calculate total magnification using ocular and objective lenses, and determine field of view diameter and area.

Uploaded by

Fat Pat
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Activity No.

03
THE MICROSCOPE

Introduction
Microscope is a powerful tool that is commonly used in laboratories to observe biological structures
and specimens that are too small to be seen with the unaided eye. It has three major parts namely:
the optical parts, the illuminating parts, and the mechanical parts. For efficient operation and use of
the microscope, one should know its parts and functions. Proper handling, manipulation, and care
of the microscope should always be observed.

At the end of this activity, students should be able to:


1. Identify the parts of a compound light microscope and explain their respective functions;
2. Describe the proper care, handling, and use of a microscope;
3. Define and calculate the total magnification for a given set of ocular and objective lenses; and
4. Determine the diameter and area of field of view.

Materials
Compound light microscope
Lens paper
Prepared slide of the letter “e”
Transparent plastic ruler

Procedure
A. Parts and Functions of a Compound Light Microscope
1. Borrow a compound light microscope from the biology stockroom.
2. Carefully carry the microscope to your workbench. Place the microscope on the table at
least five (5) inches from the edge. Remove the dust cover.
3. Identify the optical, illuminating, and mechanical parts of the microscope. State the
function(s) of each component part. Complete Figure 3.1 and Table 3.1.

B. Using a Compound Light Microscope


1. Begin by cleaning the ocular and objective lenses with lens paper.
2. Plug in the microscope and turn on the illuminator (light source).
3. If not in position, rotate the nosepiece until the scanning objective (scanner; 4x) is in place.
4. If you are using a binocular microscope, adjust the distance between the oculars to match
the distance between your eyes. If your microscope is monocular, keep both eyes open
when viewing a specimen.
5. Place a prepared slide of the letter “e” on the stage. The “e” should be positioned directly
below the scanner in its natural orientation (i.e., its tail is oriented towards you).
6. Rotate the coarse adjustment knob until “e” comes into focus. You may adjust the iris
diaphragm to achieve a good image contrast. Observe the “e” and take note of its orientation
under the microscope. Draw the letter “e” in its natural orientation as seen by the naked eye
and the image of “e” as seen under the scanner. Use the space provided in your Answer
Sheet.
7. Move the prepared slide of the letter “e” to your left. Take note of the direction of
movement of the image. Repeat this step by moving the slide to your right, away from you,
and towards you. Record your observations in your Answer Sheet.
C. Determining the Total Magnification
Using a two-lens system, a compound microscope produces an enlarged image of a specimen.
This set of lenses includes the ocular lens and the objective lens. The magnifying power of each
lens is engraved on them. Typically, an ocular lens has a 10x magnifying power. The objective
lenses also differ and can be identified from each other based on their magnifying power,
namely: scanning objective (scanner; 4x), low power objective (LPO; 10x), high power
objective (HPO; 40x), and oil immersion objective (OIO; 100x). The marking “10x” means that
the lens system generates an image ten times larger than the object being viewed. To calculate
the total magnification, simply multiply the magnifying power of the ocular lens by the
magnifying power of the objective lens.

1. Determine the total magnification of the letter “e” under the scanner, LPO, HPO, and OIO.
2. Record your answers in Table 3.2. Show your computations on the space provided.

D. Measuring the Size of the Field of View


Field of view or field of vision (FOV) is the diameter of the field (circular area) that you can
see through the microscope for a given magnification. It can be used to estimate the size of the
specimen under observation. If you know the diameter of the FOV at one magnification, you
can determine the diameter of the FOV at another magnification using the following formula:

(𝐹𝐹𝐹𝐹𝐹𝐹)(𝐹𝐹𝐹𝐹𝐹𝐹 ) = (𝐹𝐹𝐹ℎ𝐹𝐹ℎ )(𝐹𝐹𝐹ℎ𝐹𝐹ℎ )

where: FOVlow is the diameter of FOV of the low power objective


Maglow is the magnification of the low power objective
FOVhigh is the diameter of FOV of the high power objective
Maghigh is the magnification of the high power objective

For example, if 4.45 mm is the diameter of FOV at 4x magnification (scanner), then the
diameter of the FOV at 10x magnification (LPO) will be:

4.45 𝑚𝑚)(4) = (𝑚𝑚𝑚ℎ𝑚𝑚ℎ )(10) → 𝐹𝐹𝐹ℎ𝐹𝐹ℎ = 1.78 𝐹𝐹

The circular area of the FOV can be computed using the area of circle as follows:

𝐹𝐹𝐹𝐹 𝐹𝐹 𝐹𝐹𝐹 = (𝐹)(𝐹𝐹𝐹𝐹𝐹𝐹2 )

where: 𝑚 = 3.1416

Using the example above, the area of FOV under a 4x scanning objective with an FOV diameter
equal to 4.45 mm will be:

𝐹𝐹𝐹𝐹 𝐹𝐹 𝐹𝐹𝐹 = (3.1416)(2.2252 ) = 15.553 𝐹𝐹2

1. Obtain a transparent plastic ruler with millimeter (mm) metric markings. Place the ruler on
the stage and under the stage clips.
2. Carefully rotate the nosepiece until the scanner is aligned.
3. Focus up and down until the metric markings on the ruler are clear.
4. Align the ruler to measure the diameter of the FOV. The space between each marking on
the ruler should represent a 1-mm interval. Record in Table 3.3 the diameter of FOV under
a scanner magnification.
5. Calculate and record in Table 3.3 the diameter of FOV under an LPO, HPO, and OIO
magnification. Show your computations on the space provided.
6. Calculate and record in Table 3.3 the area of FOV under a scanner, LPO, HPO, and OIO
magnification. Show your computations on the space provided.

Answer Sheet 02
THE MICROSCOPE

Group No.: __________ Section: __________ Score: __________


A. Parts and Functions of a Compound Light Microscope

Figure 3.1. Compound Light Microscope.

Table 3.1. Parts and functions of a compound light microscope.


See Classification
numbered (Optical,
Function
parts in Illuminating,
Figure 3.1 Mechanical)
[1]

[2]

[3]

[4]

[5]
[6]

[7]

[8]

[9]

[10]

[11]

[12]

[13]

[14]

B. Using a Compound Light Microscope

1. Orientation of the image

Letter “e” as seen by the Letter “e” under the


naked eye scanning objective

2. Direction of movement of the image


Direction of Slide Direction of Image
a. To your LEFT
b. To your RIGHT
c. AWAY from you
d. TOWARDS you

C. Determining the Total Magnification and Size of Field of View


Table 2.2 Total magnification and size of field of view.
FOV
Objective Objective Ocular Total FOV Area
Diameter
Lens Magnification Magnification Magnification (mm2)
(mm)
Scanner
LPO
HPO
OIO

Computation for Total Magnification Computation for FOV Diameter and Area
Scanner

LPO

HPO

OIO

REVIEW QUESTIONS
1. Identify whether the following statements pertaining to the proper care of a microscope are
true or false.
a. Always carry the microscope using both hands.
b. Always carry the microscope in an upright position.
c. Start with the highest power objective when focusing a microscope.
d. Put away the microscope with the lowest power objective in position.
e. Always adjust the knob in a downward direction when focusing the objective.

2. In a properly aligned microscope, objectives are said to be parfocal. What does it mean?
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3. When should the high power objective be used? How about the low-power objective?
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