LUANAR – NRC | Biology Practical 2024 | Mark Peter

Observation of Mounted Slides using Microscope

Introduction:

Microscopy is a technique used to view objects too small to be seen with the naked
eye. Microscopy, the practice of using microscopes to view samples, is a fundamental
technique in biology. It allows for the examination of cells, tissues, and microorganisms,
providing insights into their structure and function. Proper use of the microscope is
crucial for obtaining clear and accurate images. Microscopes are essential instruments in
the field of biology, enabling scientists and students to observe objects that are too
small to be seen with the naked eye. The most commonly used microscope in
educational settings is the compound light microscope. This type of microscope uses a
combination of lenses to magnify the image of a specimen, allowing detailed
observation of its structure.

A compound light microscope consists of several key parts: the eyepiece (ocular lens),
objective lenses, stage, light source, diaphragm, and focusing knobs. The eyepiece
typically magnifies the image 10 times (10x), while the objective lenses can vary in
magnification, commonly ranging from 4x to 100x. The total magnification is calculated
by multiplying the magnification of the eyepiece by that of the objective lens.

Mounted slides are pre-prepared slides that have a specimen affixed to them, ready for
observation. There are two main types of mounted slides, permanent and temporary
slides. Permanent slides are prepared using a process that preserves the specimen
indefinitely. They are often used for educational purposes and can include samples such
as plant tissues, animal cells, and microorganisms. Permanent slides are sealed with a
coverslip to protect the specimen and ensure clarity during viewing. Temporary slides
are prepared for short-term use and are not sealed permanently. They are often used
for quick observations and are typically discarded after use. Temporary slides are useful
for observing live specimens or for quick demonstrations.

The purpose of this practical is to introduce students to the use of the compound light
microscope and to develop their skills in observing and analysing biological specimens

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using pre-prepared mounted slides. By engaging in this practical, students will learn how
to handle and operate a microscope, understand the principles of magnification and
resolution, and gain experience in scientific observation and documentation. The
practical aims to enhance students' understanding of the structure and function of the
compound light microscope, provide hands-on experience in using the microscope to
observe biological specimens, develop students' ability to make detailed observations
and record their findings accurately and foster an appreciation for the microscopic
world and its significance in biological studies.

Objectives

By the end of this practical, students should be able to:

i. Identify and describe the parts of a compound light microscope.

ii. Properly handle and use a microscope.
iii. Observe and record observations of the specimen under different magnifications.
iv. Calculate the total magnification used during observations.
v. Develop a scientific report based on their observations.

Materials

• Compound light microscope

• Prepared slides (e.g., onion epidermis, cheek cells etc)
• Lens paper
• Lab notebook

Methods

1) Carefully remove the microscope from its storage case and place it on a flat,
stable surface such as a lab bench or table.
2) If your microscope has an electric light source, plug it into a power outlet. If it
uses a mirror, ensure there is adequate ambient light.
3) Identify microscope parts.
a. Eyepiece (Ocular Lens): The lens you look through, usually with a
magnification of 10x.

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 b. Objective Lenses: Located on the rotating nosepiece, these lenses range in
magnification (e.g., 4x, 10x, 40x, 100x).
c. Stage: The flat platform where you place your slides. It has clips to hold
the slide in place.
d. Light Source: Either a built-in lamp or a mirror that reflects external light.
e. Diaphragm: Adjusts the amount of light that reaches the specimen.
f. Coarse and Fine Focus Knobs: Used to bring the specimen into focus. The
coarse focus knob moves the stage up and down significantly, while the
fine focus knob makes smaller adjustments.
4) Place the prepared slide on the stage and secure it with the stage clips.
5) Rotate the nosepiece to select the lowest magnification objective lens (usually
4x).
6) Look through the eyepiece and use the coarse focus knob to bring the specimen
into general focus. Use the fine focus knob to sharpen the image.
7) Adjust the diaphragm to control the amount of light passing through the
specimen. Too much light can wash out the image, while too little can make it
hard to see details.
8) Once the specimen is in focus at low magnification, rotate the nosepiece to a
higher magnification objective lens (e.g., 10x, 40x). Use the fine focus knob to
adjust the focus as needed.
9) Multiply the magnification of the eyepiece lens by the magnification of the
objective lens. For example, if the eyepiece lens is 10x and the objective lens is
40x, the total magnification is (10 times 40 = 400) times.
10) Make detailed notes of what you observe at each magnification level. Include
descriptions of the specimen's structure, any visible cells or tissues, and any
notable features.
11) Draw what you see through the microscope, labelling key parts of the specimen.
12) Turn off the light source and unplug the microscope if necessary.
13) Use lens paper to clean the slides and coverslips.
14) Cover the microscope with a dust cover and store it in a safe place.

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 Appendix 1

Part Function
Ocular lens (Eyepiece) The lens at the top of the microscope that you look through,
is usually 10x or 15x power.
Diopter adjustment Adjusts the focus to compensate for differences in vision
between the user's two eyes.

Head Houses the optical elements of the unit, including the

eyepiece and objective lenses.
Nosepiece The rotating part that holds the objective lenses and allows
the user to change magnification levels.

Objective lens Primary optical lenses ranging from 4x to 100x

magnification.
Stage clip Clips on the stage that hold the slide in place
Aperture Hole in the stage through which light passes to illuminate the
specimen
Stage A flat platform where the slide is placed, with clips to hold
the slide in place.

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 Diaphragm Controls the amount of light that reaches the specimen.

Condenser Lens system below the stage that focuses light onto the
specimen
Illuminator (Light Source) A light source for the microscope, providing illumination for
the specimen
Arm (Carrying Handle) Connects the head to the base and is used to carry the
microscope
Mechanical stage Stage that can be moved precisely with knobs for accurate
positioning of the slide
Coarse adjustment Larger adjustment knob that moves the stage up and down
to bring the specimen into focus
Fine adjustment Smaller adjustment knob that allows for finetuning the focus,
especially at higher magnifications.

Stage controls Knobs that move the mechanical stage left/right and
forward/backward to position the slide

Base The bottom part of the microscope that provides stability

and houses the illuminator
Brightness adjustment Control that adjusts the intensity of the light source to
enhance the visibility of the specimen
Light switch Switch that turns the illuminator on and off.

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