Republic of the Philippines

President Ramon Magsaysay State University

(Formerly Ramon Magsaysay Technological University)
Iba, Zambales, Philippines
Tel/Fax No.: (047) 811-1683

College/Department College of Arts and Sciences

Course Code CASNS 1a
Course Title General Botany
Place of the Course in the Program Major (Core) Course
Semester & Academic Year Second Semester, AY 2020-2021

Laboratory Module 1
THE MICROSCOPE
Introduction
The world of Biology is everywhere. Microscopic organisms are teeming in areas that we
may not expect them to be. However, our naked eyes – a normal eye with regular vision unaided by
any tools – cannot see objects that are smaller than 0.1 millimeter in size. Because of this, the world
was surprised to learn that smaller organisms such as the Euglena and Paramecium may be seen
thriving in a community as small as a drop of water. In this module, you will learn about the tool
that advanced the studies in science especially in Biology.

Intended Learning Outcomes

At the end of this laboratory module, the student should be able to:
1. Acquire the knowledge about the history of a microscope;
2. Determine the different parts and functions of the microscope;
3. Calculate magnification; and
4. Understand the proper handling and care for a compound microscope.

Discussion
A. Microscope – Its Word Origin
A microscope is described as "optical instrument which by means of a lens or lenses
magnifies and renders visible minute objects or details of visible bodies”. Galileo submitted to the
Accademia dei Lencei in 1624 and called it “occhiolino” or “litte eye”. The term microscope was
coined by Giovanni Faber in 1625 as an analogy to telescope.
B. Timeline of a Microscope

 14th century: spectacles first made in Italy

 1590: Two Dutch spectacle-makers and father-and-son team, Hans and Zacharias Janssen,
create the first microscope.
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  1667: Robert Hooke's famous "Micrographia" is published, which outlines Hooke's various
studies using the microscope.
 1675: Enter Anton van Leeuwenhoek, who used a microscope with one lens to observe insects
and other specimen. Leeuwenhoek was the first to observe bacteria. 18th century: As
technology improved, microscopy became more popular among scientists. Part of this was
due to the discovery that combining two types of glass reduced the chromatic effect.
 1830: Joseph Jackson Lister discovers that using weak lenses together at various distances
provided clear magnification.
 1878: A mathematical theory linking resolution to light wavelength is invented by Ernst
Abbe.
 1903: Richard Zsigmondy invents the ultramicroscope, which allows for observation of
specimens below the wavelength of light.
 1932: Transparent biological materials are studied for the first-time using Frits Xernike's
invention of the phase-contrast microscope.
 1938: Just six years after the invention of the phase contrast microscope comes the electron
microscope, developed by Ernst Ruska, who realized that using electrons in microscopy
enhanced resolution.
 1981: 3-D specimen images possible with the invention of the scanning tunneling microscope
by Gerd Binnig and Heinrich Rohrer.

C. Parts of a Microscope
A typical microscope is constructed of several very important pieces that you will need to be
aware of during general use.

 Eyepiece: These lenses, also known as ocular lenses, are typically 10x, but also come in 5x,
15x and 20x varieties. The eyepiece lens is what you actually look through to see your
specimen. The interocular distance is adjustable so that you can keep both eyes open when
looking into the microscope.

 Nosepiece: Also called a revolving nosepiece or turret, this structure holds the objective
lenses. Rotating this allows the objective lens to be switched.

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 Objective lenses: The lens closest to the object being viewed is known as the objective lens.
Each objective lens has a different magnification including scanning lens (10x), mid-power
lens (40x), and high-power lens (100x).

 Stage: This area is the main, flat plate that holds the slides for observation.

 Diaphragm: Most useful at higher powers, the diaphragm is typically a five-hole disc
housed under the stage to adjust the amount of light passing through the stage opening. Each
opening is a different diameter controlling how much light passes through to help properly
illuminate the specimen while also increasing contrast and resolution.

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 Base: The base is the bottom support structure of the microscope. To carry a microscope,
grip the arm with one hand and place the other hand underneath the base.

 Body: The body refers to the main section of the microscope minus the stand (base) or any
illuminators. The body components generally include the eyepiece and objective lenses but
not the focusing block.

 Arm: The arm is the area of the microscope between the tube and the base. To carry a
microscope, grip the arm with one hand and place the other hand underneath the base.

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 On/Off Switch: Controls the power to the microscope.

 Coarse Focus: The larger of two adjustment knobs that moves the objective lenses closer or
farther away from the specimen in large steps.

 Fine Focus: The smaller of two adjustment knobs moves the objective lenses closer or
farther away from the specimen in very small steps. It can be used to fine tune the focus on
various parts of a specimen after first using the Coarse Focus to get close.

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  Stage Adjustment Knob: The mechanical stage allows for adjustment of the slide position
forward, backward and left to right.

D. Microscope Lenses
One of the most important parts of the microscope are the lenses used to actually view the
specimen. The microscope uses the eyepiece lens, or the lens you look through, to present the
image of the specimen. Eyepiece lenses are typically 10X, but may also come in 5X, 15X or 20X.
The power is generally found labeled on the eyepiece itself.

The lens nearest the object being viewed is called the objective lens. These lenses can come in a
range of powers such as 4X ( Scanning ), 10x ( Low Power ), 40X ( High Power ) or 100X ( Oil
Immersion Objective ). Some lenses, such as the 100X lens, are specifically designed to be used
with immersion oil.

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The power of each lens can be found labeled on the side of the tube housing. Additionally, each lens
has a specific, color-coded band that denotes its resolution: red for 4X, yellow for 10X, blue for
40X and white for 100X.

E. Immersion Oil Lenses

Most lenses are used ‘dry’ or without oil. Some
lenses use a special type of oil to increase their
resolution. Oil is typically used with higher powered
lens such as 100X. These lenses will typically have
special markings on the side of the housing to indicate it
is designed to be used with oil.

When used with oil, the lens is lowered until it just contacts a small drop of oil placed on the
specimen slide. The oil serves as a bridge between the lens and the glass plate containing the
specimen helping to focus the light and increase the resolution. Once oil is added to a slide, the
objective lens should not be switched back to a non-oil lens.

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 Normally, some of the light is lost due to refraction when using the lens without immersion
oil. The addition of immersion oil helps to refocus the light as it passes from the slide to the lens.
This provides an image with better resolution and contrast.

F. The systems of a microscope

The compound microscope has two systems of lenses for greater magnification, 1) the
ocular, or eyepiece lens that one investigates and 2) the objective lens, or the lens closest to the
object. Before purchasing or using a microscope, it is important to know the functions of each part.

G. Magnification
Magnification means the numbers of times an object is enlarged by a lens system or
enlarged or reduced in the drawing or illustration.
In the microscope, magnification is achieved by a system of curved lenses of the ocular and
objectives. The ocular further magnifies the image formed by the objective so that the image seen
by the eye has a magnification equal to the product of the magnification of the two lens systems.
This is known as the linear magnification.
When you don’t know the magnification of the eyepiece and objective used:
Magnification = (Image size) / (Actual size)
 Image size = size on the paper
 Actual size = size an object really is

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When you know the magnification of the eyepiece and objective used:

H. Microscope Care
Microscope is a very sensitive instrument and costs a lot of money. A compound
microscope is what usually being use in the schools and university by science-major students.
Proper handling and care for microscopes are advised to students to avoid accidents and to avoid the
units to become dysfunctional in the long run.

1. When moving and setting up a microscope, proper care should be given to how the device is
held. Remove the microscope from the cabinet by gripping the arm firmly with one hand
while placing the other hand underneath the base of the device for support. Do not lift on the
stage. Do not remove the dust cover until the microscope is placed and positioned on the
table where it will be used.

2. Never touch the lenses with your fingers. Oil from your hands can smudge the lens and in
some cases even leave scratches. If you must clean the lens, use only the appropriate lens
tissues to avoid damaging the lens.

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 3. When your work with the microscope is finished, remember to rotate the nosepiece back to
the lowest power objective. Lower the stage to its lowest position. Make sure the power cord
is neatly wrapped up and replace the dust cover before returning the microscope to the
cabinet.

4. When you finished using the microscope:

a. Make sure that the stage manipulator bar is not extended out to the left.
b. Unplug the microscope and coil the cord beside the microscope.
c. Put the dust cover back on the microscope.
d. Carefully slide the microscope back into place.

Suggested Readings/Videos
1. History and Importance of Microscope - https://www.youtube.com/watch?v=uAUQ_1SH-
RY
2. Microscope for Beginners - https://www.youtube.com/watch?v=eZX9U15F5Q8
3. Microscope Setup and Use - https://www.youtube.com/watch?v=SUo2fHZaZCU
4. Magnification Calculation - https://www.youtube.com/watch?v=FdaLMkoHF2o
Activity
1. Draw the compound microscope and label its parts.
2. Print the image below (do not adjust the format of this document to avoid discrepancy in the
size of the photo). Answer the questions in your worksheet about calculating magnification.

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Name: ____________________________ Score: _____
Course and Year: ___________________

Laboratory Worksheet No. 1

THE MICROSCOPE

The Compound Microscope

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 I. Answer the following problem:

The actual size of the fly’s eye is 1,000 μm. What is the magnification?

a. Length of eye is ____________mm.

b. _______ mm x 1000 = μm
c. Image size = __________ μm
d. Magnification = Image / Actual
i. Magnification = ________ μm / ________ μm
ii. Magnification = _____________
iii. The picture shows the eye magnified (zoomed in) by __________x.

Reflection
1. What do you think is the importance of microscope in Botany?
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