LAB MANUAL ALL

MATRICULATION
DIVISION

BIOLOGY

LABORATORY MANUAL
SEMESTER I & II
DB014 & DB024

FOURTH EDITION

MATRICULATION DIVISION
MINISTRY OF EDUCATION MALAYSIA

BIOLOGY

LABORATORY MANUAL
SEMESTER I & II
DB014 & DB024

MINISTRY OF EDUCATION MALAYSIA
MATRICULATION PROGRAMME
FOURTH EDITION

First Printing, 2011 (First Edition)
Second Printing, 2015 (Second Edition)
Third Printing, 2018 (Third Edition)
Fourth Printing, 2020 (Fourth Edition)
Copyright © 2020 Matriculation Division
Ministry of Education Malaysia

ALL RIGHTS RESERVED. No part of this publication may be reproduced
or transmitted in any form or by any means, electronic or mechanical,
including photocopying, recording or any information storage and retrieval
system, without the prior written permission from the Director of
Matriculation Division, Ministry of Education Malaysia.

Published in Malaysia by

Matriculation Division
Ministry of Education Malaysia,
Level 6 – 7, Block E15,
Government Complex Parcel E,
Federal Government Administrative Centre,
62604 Putrajaya,
MALAYSIA.
Tel : 603-88844083
Fax : 603-88844028
Website : http://www.moe.gov.my/v/BM

Printed in Malaysia by

Malaysia National Library
Biology Laboratory Manual
Semester I & II
DB014 & DB024
Fourth Edition

ISBN 978-983-2604-41-9

NATIONAL EDUCATION PHILOSOPHY
Education in Malaysia is an on-going effort towards further
developing the potential of individuals in a holistic and
integrated manner, so as to produce individuals who are
intellectually, spiritually and physically balanced and
harmonious based on a firm belief in and devotion to God.
Such an effort is designed to produce Malaysia citizens who
are knowledgeable and competent, who posses high moral
standards and who are responsible and capable of achieving a
high level of personal well-being as well as being able to
contribute to the betterment of the family, society and the
nation at large.

NATIONAL SCIENCE EDUCATION PHILOSOPHY

In consonance with the National Education Philosophy, science
education on Malaysia nurtures a science and technology
culture by focusing on the development of individuals who are
competitive, dynamic, robust and resilient and able to master
scientific knowledge and technological competency.

iii

FOREWORD

I am delighted to write the foreword for the Laboratory Manual,
which aimed to equip students with knowledge, skills, and the
ability to be competitive undergraduates.

This Laboratory Manual is written in such a way to emphasise
students’ practical skills and their ability to read and understand
instructions, making assumptions, apply learnt skills and react
effectively in a safe environment. Science process skills such as
making accurate observations, taking measurement in correct
manner, using appropriate measuring apparatus, inferring,
hypothesizing, predicting, interpreting data, and controlling
variables are further developed during practical session. The
processes are incorporated to help students to enhance their
Higher Order Thinking Skills such as analytical, critical and
creative thinking skills. These skills are crucial to prepare students
to face upcoming challenges in the 21st century era.

The manipulative skills such as handling the instruments, setting
up the apparatus correctly and drawing the diagrams can be
advanced through practical session. The laboratory experiments
are designed to encourage students to have enquiry mind. It
requires students to participate actively in the science process
skills before, during and after the experiment by preparing the pre-
report, making observations, analysing the results and in the
science process skills before, during, after the experiment by
preparing the pre-report, making observations, analysing the
results and drawing conclusions.

It is my hope and expectation that this manual will provide an
effective learning experience and referenced resource for all
students to equip themselves with the skills needed to fulfil the
prerequisite requirements in the first-year undergraduate studies.

Dr. HAJAH ROSNARIZAH BINTI ABDUL HALIM
Director
Matriculation Division

iv

CONTENTS Page

Foreword iv
Content v
Learning Outcomes vii
Introduction x

SEMESTER I 1
4
Experiment Title 11
1 Basic Techniques In Microscopy 17
2 Animal Tissues 18
3 Plant Tissues 20
4 Cell Division – Mitosis
5 Pollen Germination
6 Plant Growth

v

SEMESTER II 22
24
Experiment Title 27
7 Diversity of Bacteria 36
8 Plant Diversity: Gymnosperm 39
9 Animal Diversity: Invertebrates and Vertebrates 45
10 Chloroplast in Aquatic Plant
11 Homeostasis 53
12 Coordination 54

References
Acknowledgements

vi

1.0 Learning Outcomes

1.1 Matriculation Science Programme Educational Objectives

Upon a year of graduation from the programme, graduates are:

1. Knowledgeable and technically competent in science
disciplines in-line with higher educational institution
requirement.

2. Able to communicate competently and collaborate
effectively in group work to compete in higher education
environment.

3. Able to solve scientific and mathematical problems
innovatively and creatively.

4. Able to engage in life-long learning with strong
commitment to continue the acquisition of new knowledge
and skills.

1.2 Matriculation Science Programme Learning Outcomes

At the end of the programme, students should be able to:

1. Acquire knowledge of science and mathematics
fundamental in higher level education.
(PEO 1, MQF LOD 1)

2. Demonstrate manipulative skills in laboratory work.
(PEO 1, MQF LOD 2)

3. Communicate competently and collaborate effectively in
group work with skills needed for admission in higher
education institutions.
(PEO 2, MQF LOD 5)

4. Apply logical, analytical and critical thinking in scientific
studies and problem solving.
(PEO 3, MQF LOD 6)

5. Independently seek and share information related to science
and mathematics.
(PEO 4, MQF LOD 7)

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1.3 Biology 1 Course Learning Outcome
At the end of the course, student should be able to:

1. Explain the main concepts and theories in cells, biomolecules,
inheritance, genetics and biological development. (C2, PLO
1, MQF LOD 1)

2. Conduct basic biology laboratory work on microscopy,
tissues, genetics information, pollen germination and plant
growth by applying manipulative skills. (P3, PLO 2,
MQF LOD 2)

3. Solve basic problems related to cells, biomolecules,
inheritance, genetics and biological development.
(C3, PLO 4, CTPS1, MQF LOD 6)

1.4 Biology 2 Course Learning Outcome
At the end of the course, student should be able to:
1. Explain the basic concepts and theories in transport system
processes, mechanisms for adaptations in living things,
ecological and environmental issues in biology. (C2, PLO 1,
MQF LOD 1)

2. Conduct basic biology laboratory work on diversity of
living things, chloroplast in aquatic plant, homeostasis
and coordination by applying manipulative skills. (P3,
PLO 2, MQF LOD 2)

3. Solve basic problems related to transport system
processes, mechanisms for adaptations in living things,
ecological and environmental issues in biology. (C3,
PLO 4, CTPS 3, MQF LOD 6)

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1.5 Biology Practical Learning Outcomes
Biology experiment is to give the students a better
understanding of the concepts of Biology through experiments.
The aims of the experiments in this course are to be able to:
• know and practice the necessary safety precautions to be
taken.
• use the correct techniques of handling apparatus.
• plan, understand and carry out the experiment as
instructed.
• observe, measure and record data consistency, accuracy
and units of the physical quantities.
• define, analyse data and information in order to evaluate
and deduce conclusions from the experiments.
• discuss data and information logically and critically.
• analyse and draw conclusions from biological data.
• develop solution to biological problems.
• acquire scientific skills in measuring, recording and
analysing data as well as to determine the uncertainties
(error) in various physical quantities obtained in the
experiments.
• understand the limitations to the accuracy of observations
and measurements.

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INTRODUCTION

A. General Guidelines

Laboratory Regulation

1. Always wear laboratory coats and covered shoes in the lab.
2. Do not eat or drink in the laboratory.
3. Use the apparatus and materials wisely.
4. Do not throw rubbish and residues into the sink. Wrapped and

throw them into the dustbin provided.
5. At the end of the experiment, students must

a) clean the apparatus using the detergent provided.
b) soak the apparatus in acidic solution containing mild

hydrochloric acid.
c) wash the sink and work station.
d) make sure that all the tables are clean and neat.
e) place the materials and apparatus in their respective places.

Sectioning and Staining Plant Tissues

1. Sectioning of plant tissues or parts must be made and stained before
they are examined under the microscope.

2. Use sharp blade or microtome to make a thin slice of the specimen.
3. Clean the blades with water and dry them using tissue paper after

being used.

Preparation for Experiment

1. You are advised to read the manual before carrying out the
experiment. You are also advised to make additional references
about the topic.

2. Prepare a rough layout of the experiment that consists of tables,
graphs and space for drawing.

3. Identify the equipments and materials that are going to be used in
the experiment. This will maximise the time used for experiment.

4. Follow strictly the instructions in the manual.
5. Record only what you observe in the experiment.

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Laboratory Report and Evaluation

1. Report should contain the following:-

Title
Objective(s)
Introduction (hypothesis/variable/problem statement)
Procedures (in passive voice, past tense, in reporting style)
Observation (tables, graphs, data, drawing)
Analysis / Discussion regarding tables, graphs, data or drawings
Conclusion
Questions
References

2. Reports must be handwritten or typed.
3. Diagrams should be drawn on the blank sheet using a 2B pencil.

All diagrams must be labelled.
4. Metric system must be used in writing numerical data.
5. Data can be presented in the form of graphs, tables, flow charts

or diagrams. Give suitable titles to the graph, table, flow charts
and diagrams.
6. Record the following on the front page of the report.

College’s name:
Student’s name:
Matriculation number:
Practicum group:
Title:
Date:
Tutor’s / lecturer’s name:

7. Submit your report to your lecturer at the end of the practical
session.
The report and the attendance for each lab will be evaluated and
included in the assessment.

Scientific Drawing

1. Diagrams drawn must be based on the observation of the specimen
and not copied from books.

2. All parts of the specimens observed must be drawn using the right
scale.

3. An overall drawing or plan drawing must be made to show the parts
where the drawings are made.

4. Show clear orientation of the specimen so that the position and the
relationship with other organs can be determined.

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5. Use a sharp 2B pencil to draw thin, clear and continuous lines.
6. Drawing must not be coloured or shaded to differentiate the

systems from tissues. For this purpose, students are allowed to use
various patterns to differentiate systems.
7. Label all your drawings. All labels must be written on the right and
left side of the diagram. Do not write the labels on or in the diagram.
Labels must be written horizontally. Straight line must be used to
connect the structure.
8. Magnification used in the drawing from observation under the
microscope must be mentioned; e.g.: 40x or 100x actual
magnification.
Caring for Plants and Animals
1. Water the plants every day. Make sure the soil is damp and wet.
2. Clean the animal cages every day. Make sure the cages are in good
condition.
3. Feed the animal daily.

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B. Introduction to Microscopy

The discovery of microscope started a new era in biology since for the
first time man was able to observe cells, the basic units of life.

The optical properties of lenses have been known for the last 300 years
B.C., but these knowledge were not used to the fullest until the
seventeenth century when Antonio Van Leeuwenhoek (1632-1732), a
Dutch, and his colleagues discovered a simple workable microscope.
With the discovery of the simple microscope, many people were able to
observe minute living organisms in great details. One of them was
Robert Hooke who in 1665 gave the first extensive description of his
experience in observing cork tissue using the simple microscope. This
marked to the beginning of the study of cells. Below is the excerpt from
the journal Micrographia by Hooke of what he observed from the cork
tissue under the microscope:

“… I could exceedingly plainly perceive it to be all perforated and
porous….. these pores, or cells, ….. were indeed the first
microscopical pores I ever saw, and perhaps, that were ever seen,
for I had not met with any writer or person, who has made any
mention of them before this”.

Although the description by Hooke about the cork tissue might sound
hilarious, you may have described them in the same way had you lived
in the seventeen century when the concept of cell as the fundamental
unit of life was something unknown.

1. What is a Microscope?

Microscopes are precision instruments, and therefore need to be handled
carefully. Many people think that microscopes can only be used to
observe objects in higher magnification. If a microscope can only be
used to observe a magnified image, then its usage is limited. In fact,
microscope can be used to magnify an object, determining the size of an
object and observing fine details of an object, all of which are not
discernible to our naked eyes. Therefore, before one can properly use a
microscope, first he has to be familiar with the microscope and be able
to identify the components of the microscope and their functions.

With the advancement of technology in microscopy, many high-quality
microscopes have been designed for many specific uses. Nowadays,
many microscopes are of the compound types which use two sets of
lenses. The first set of lens constitutes the objective lens which supplies
the initial real magnified image. The second set of lens constitutes the
ocular lens which magnifies further the image formed by the first set of

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lens and converts the real image into virtual image which is in turn
viewed by the user’s eyes. In compound microscopes, the actual

magnification is calculated as the magnification of objective lens

multiplied by the magnification power of the ocular lens.

Today there are many types of light microscope, for example the phase-
contrast microscope that allows user to view living cells or specimens
without the use of stains to increase the contrast. Contrast is based on
the differential absorption of light by parts of the specimen. There are
compound microscopes, which employ ultraviolet light as the source of
light, making it possible to view specimens that emit fluorescence. Such
microscopes are now commonly used in diagnostics laboratories and
research. There are also other compound microscopes which use either
dark field or light field. Another type of microscope is compound
microscope with inverted objective, called inverted microscope, which
is used to observe living cell cultures.

A microscope is not only capable of producing the image of an object
but also capable of distinguishing between two adjacent points on the
object. This capacity is termed as the resolving power of the lenses or
the resolving power of the microscope. The higher the resolution of the
microscope, the higher is the ability to distinguish details of the object.
Microscope quality depends upon the capacity to resolve, not magnify,
objects. Magnification without resolving power, however, is not
worthless in the field of microscopy.

The resolving power of a light microscope depends upon the wavelength
of light (colour) being used, and not on a value called the numerical
aperture (N. A) of the lens system used. The numerical aperture is
derived from a mathematical expression that relates the light transferred
to the specimen by the condenser to the light received by the objective
lens. This relationship is given by the following expression:

Resolving power = Shortest diameter of the
observed structure

= Wavelength ()
Numerical Aperture (N.A)

Thus, the resolving power is increased by reducing the wavelength of
the light used. The shorter the wavelength used, the shorter will the
diameter of the structure being observed, or in other words, the resolving
power is increased. The resolving power cannot be increased
substantially because the light spectrum is narrow (500 nm). However,
we can increase the resolving power by increasing the numerical
aperture in the lens system of the microscope. When the specimen is

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illuminated with light from direct or oblique direction, the relationship
is given as follows:

Resolving power = Wavelength ()
2 x N.A

where

λ - wavelength of light
N.A - Numerical Aperture

The condenser located below the mechanical stage or slide holder can
transfer oblique and direct light sources to the specimen and this can
approximately double the numerical aperture (N.A). Thus, the resolving
power can be increased. Therefore, the condenser has to be properly
focused to achieve high resolving power.

Light enters the specimen, and some of it will be refracted as it goes
through the air. This light will not enter the objective lens. By placing
oil of immersion in the space between specimen and objective lens, we
can reduce the light refraction and increase the amount of light entering
the objective lens resulting in a brighter and clearer image. The oil of
immersion used should have the same refractive index (R.I) as the glass
to reduce refraction.

After understanding some principles of microscopy, we need to identify
the components of the microscope and know their functions. The
microscope to be used in the laboratory is the bright field light
compound. The diagram of the microscope is shown in Figure 1.
Familiarise yourself with a microscope and its functions before using it
(refer Table 1).

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Figure 1: Compound light microscope

Each objective lens will generate an image in a specific field of view.
The size (diameter) of the field of view depends on the type of
objective lens used. As the magnifying power of the objective lens
increases, the size of the field of view decreases, and the working
distance, the distance between the slide and the objective lens, also
decreases. When the specimen field of view is wide, more light will
enter the objective lens, so it is important to regulate the amount of light.
Figure 2 shows the relationship between objective lens, fields of view
and working distance for each of the objective lens.

Figure 2 Comparison of working distance at three different objective magnification

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Table 1 Components of microscope and their functions

Component Function

1a. Ocular lens or eyepiece lens: 1a. Magnifies the real image and
This lens is found at the top of the converts it to a virtual image to be
microscope. It normally has a viewed by user’s eyes.
magnification power of 10x or 15x.
1b. To adjust and compensate the
1b. Ocular control knob. differences in binocular image of the
eyes.

2. Body tube. Body tube is the hollow housing which
supports the ocular lenses at the top and
connects them with the objective lenses
below it.

3. Rotating nosepiece: The structure to which the objective
You will hear clicking sounds when the lenses are mounted. By gently rotating
objective lens is in its correct position the nosepiece, you may choose the
above the specimen. You may practise objective lens you want and correctly
this by rotating and changing the place it over the specimen.
objective lenses.

4. Objective lenses:
Normally there are 3-4 objective lenses
mounted on the nosepiece, and these can
be rotated and changed as you require.

a. Scanning objective lens (4x): a. Used to scan the specimen before
Coarse specimen field = 5 mm identifying the specific part to be
viewed further.

b. Low-power objective lens (10x): b. Used to view major part of the
Small specimen field = 2 mm specimen.

c. High-power objective lens (40x): c. Used to view specific part of the
Very small specimen field = 0.5 mm specimen.

d. Oil immersion objective lens (100x):
Immersion oil is placed in the space

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between the specimen and objective lens d. Used to view microorganisms such

to reduce the light refraction from the as bacteria and microstructures in the

specimen. cells.

Very small specimen field = 0.2 mm

5a. Stage: a. The horizontal surface on which a
The horizontal surface which has a hole specimen is placed.
in the centre to allow light from below
to focus on the specimen.

5b. Slide clips. b. The stage is usually equipped with
slide clips to hold the slide in place.

5c. Slide adjustment knob c. Two knobs are used to move the slide
to the left, right, forward or
backward. Move these knobs to
learn how the slide is moved into
position.

6a. Condenser: a. Used to focus and deliver light to the
Condenser is located immediately under specimen.
the stage
b. Used to adjust the condenser.
6b. Condenser control knob. c. Used to focus the light.
6c. Condenser lens knob.

7. Iris diaphragm: Controls the amount of light entering
An adjustable light barrier of iris type and leaving the condenser.
built into condenser. The size of the
diaphragm is controlled by rotating the
knob either to the left or right. Rotate
the knob to the left and to the right and
observe what happens.

8a. Off/on switch. The source of light is a tungsten bulb
located at the base of a microscope.
8b. Light control knob.
Please ensure that either of the switches
is OFF or MINIMUM, respectively
before you use the microscope.

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9. Body arm The metal part used to carry a
10. Base microscope.

The heavy cast metal part used as the
base and for support.

11. Coarse adjustment knob: Used to bring specimen into focus by
Use this knob only when using low- moving the stage to the specimen.
power objective lens. Rotate this knob
carefully and observe what happens.
Does the stage or the body tube move?

12. Fine focus adjustment knob Used to bring specimen into focus while
using high-power or oil immersion
objective lenses.

Now that you have become familiar with the component parts of the
microscope, you can proceed to use the microscope. Check the
microscope to ensure that it is in good working conditions.

2. Setting Up of a Light Microscope

a) Plug the microscope to a power source. Before switching on the
plug, check that the light switch is OFF or the light control knob
is set at MINIMUM

b) Switch on the power. Turn on the light control knob or adjust
the light diaphragm to deliver the light to the specimen field (but
not too much light). To focus the condenser, do the following:
i) Take a prepared slide and place it on the stage.
ii) Rotate the nosepiece and put the coarse objective lens
into position above the specimen.
iii) Move the stage upwards by rotating the coarse adjustment
knob until it stops completely.
iv) While looking through the oculars, move the stage
downwards using the fine adjustment knob until the
specimen is in focus.
v) To focus the condenser, you need to bring the specimen
and the condenser into focus in the same plane. Close
down the iris diaphragm and reduce the amount of light.

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3. Focusing a Specimen

a) Place a prepared slide on the stage (for this exercise you may
use any of the prepared slides available in the lab). Move the
slide so that the specimen is placed in the centre and under the
objective lens.

b) First you need to ensure that either the scanning objective lens
(4x) or low-power objective lens (10x) is placed above the
specimen.

c) While looking at the slide from the side, move the stage upwards
until it stops completely. Use the coarse adjustment knob to do
this.

d) Now observe the specimen through the ocular lens. The
specimen will appear blur because it is still not focused. To
focus the specimen, gently move the stage downwards until the
specimen comes into sharp focus and clear. Use the fine
adjustment knob to do this.

Look through the ocular lens with both eyes. You may see the
image differently between your right and left eyes. Do the
following to adjust the ocular lenses for the differences between
your eyes. Determine which ocular lens is adjustable. Close the
eye over that lens and bring the specimen into sharp focus for
the open eye (right eye). Open the other eye (left eye) and close
the first eye (right eye). If the specimen is still not in sharp focus,
turn the adjustable ocular control knob (1b) until the specimen
is in focus. You may now look with your eyes through both
ocular lenses.

e) After the specimen has been focused by the low-power objective
lens, rotate the nosepiece to change to the high-power objective
lens (40x). You will hear a clicking sound when the objective
lens comes into its correct position right above the specimen.
The microscope used should be of PARFOCAL type, that is
once a specimen has been focused using a particular objective
lens, it will stay focused for the other objective lenses. Using
this microscope, you do not need to refocus the specimen when
you change the objective lens. You just need to adjust the fine
focus adjustment knob.

f) If the field of view is dark or too bright, adjust the amount of
light by using the light control or diaphragm knob.

g) When you have finished using the microscope, rotate the
nosepiece to place the coarse objective lens (4x) back in position
over the centre of the stage. Remove the last slide and clean the
stage if necessary.

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4. Using Oil Immersion Objective Lenses

The oil immersion objective lens is used when you want to observe a
specimen at the highest resolution with the light microscope or when the
resolution of other objective lens is not sharp and clear enough. The
objective lens is usually used to observe microorganisms such as
bacteria and protozoa or to observe microorganelles in the cell. Before
using the objective lens, the specimen has to be fixed and stained to
increase its contrast.

a) Follow steps (a) to (d) in procedure (3).
b) Rotate the nosepiece to bring the high-power objective lens (40x)

half way as shown in Figure 3.
c) While holding the nosepiece in this position, apply a single small

droplet of immersion oil to the illuminated spot on the slide.

Figure 3 Using oil immersion objective lenses

d) Rotate the nosepiece again to move the high-power objective lens
into position until you hear a clicking sound. The objective lens
is now right above the specimen and will be immersed in the oil.

e) Open up the iris diaphragm to increase the amount of light.
f) While looking at the specimen through the ocular lens, use the

fine adjustment knob until the specimen comes into sharp focus
and become clear. If you have any problems, consult the
instructor / tutor.
g) When you have finished using the oil immersion objective lens,
do the following steps:
i) Carefully move the stage downwards.
ii) Clean the oil immersion objective lens by gently wiping it

with clean lens tissue. If the objective lens is still dirty,

xxi

clean it with a little amount of xylene and rub it gently with
clean, dry lens tissue.
iii) Remove the slide off the stage.
iv) Gently rotate the nosepiece again to place the low-power
objective lens back in position over the centre of the stage.
v) If oil is found on the stage, wipe the oil off with lens tissue
and with some alcohol.

5. Storage of Microscopes

When you have finished using the microscope, do the following to store
the microscope.

a) Check that you have not left a slide on the stage.
b) Check that the stage is clean without any trace of water or dust on

it. If there is any water on the stage, wipe it off with dry tissue. If
it is oil, wipe it off with dry tissue with some alcohol.
c) If you use oil immersion objective lens, gently wipe it with clean
lens tissue.
d) Check that the scanning objective lens (4x) is placed back in
position over the centre of the stage.
e) Turn off the light switch or close down the iris diaphragm to
reduce the amount of light to a minimum and then switch off the
power.
f) Tie up the power chord below the body arm.
g) Ensure the slide clips are placed on the stage and they are not
protruding.
h) HOLD THE MICROSCOPE WITH BOTH HANDS, that is
hold the body arm of the microscope with one hand and the base
of the microscope with the other hand.

6. The Dissecting Microscope (Stereoscopic Microscope)

The dissecting microscope (Figure 4) is used for observations at low
magnification in binocular view (involving 2 ocular lenses) or in three
dimensions. Specimens are often viewed in a fresh state and need not be
placed on a slide. The microscope is ideal for dissection of a small
specimen. The procedures of using a dissecting microscope are basically
similar to the procedures for a light microscope; however, it is simpler
to use than a light microscope.

1. Place a specimen on the specimen plate at the base (5).
2. Illuminate the specimen, by switching on the light source.

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a) The eyepiece lenses need to be adjusted to suit your eyes and
to ensure that the image remains clear at different
magnifications. Turn the adjustment knob (2) to position ‘O’

b) Adjust ocular 1 so that the two oculars fit well with both eyes.
c) Turn the magnification control knob (4) to select the

magnification to 4x.
d) Look through both ocular lens and focus the specimen by

turning the focus knob (8).
e) Change the magnification to 0.8x by turning the control knob

(4).
f) Observe the image with your right eye and focus using the

adjustment knob located on the right ocular until the image
becomes clear.
The microscope has now been adjusted to suit your eyes so that you can
take advantage of the stereoscopic effect.
3. Look through the oculars with both eyes. Focus the image by
turning the focus knob (8). Specimen as high as 20 mm may be
focused using the adjustment knob.
4. For specimen higher than 20 mm, the microscope may be focused
by moving its body (3) upwards. This is done by turning the body
screw (7) loose and moving the body upwards or downwards along
the stand (6) as far as the stop screw (9). Tighten the body screw
(7) when the body is at the right position.
5. The stop screw (9) prevents the body of the microscope from
crashing on to the specimen plate at the base.

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Figure 4 A dissecting microscope

7. Electron Microscope

This microscope makes use of the electron beams instead of light source.
Electron beams have very short wavelength of approximately 0.005 mm,
and therefore theoretically, the microscope can resolve objects as small
as 0.0025 nm in diameter. The resolution of an electron microscope is
usually 1 to 1.2 nm. With electron microscope, magnifications up to
250,000 are commonly obtained with biological materials. The shorter
wavelengths of electrons are said to have greater resolving power than
those of light microscope. There are two types of electron microscope,
namely the transmission electron microscope and the scanning electron
microscope.

In transmission electron microscope, the electron beams are used instead
of light source. An image will be formed on a photographic film screen.
The microscope uses an electromagnetic lens as a condenser and the
electron source is focused by the condenser lens through the specimen.
The image is then magnified by the objective lens and the projector lens.
An image taken from the electron microscope is called a transmission
electron micrograph. In transmission electron microscope (TEM), only
very thin sections of specimen of < 30 nm are used for microscopic
observation. They are placed on a copper grid used for support.
Electrons cannot be seen with the human eye, so the image is made

xxiv

visible by shinning the electrons on to a fluorescent screen. This will
only produce black-and-white pictures. The electron microscope can be
used only for dead tissue materials because they are viewed in vacuum.
In scanning electron microscope, specimens are coated with a heavy
metal such as gold. Electron beams will not be focused through the
specimen, and when the electron beams collide with the specimen, some
electrons will be absorbed while some are deflected or scattered. Those
parts of the specimen which are denser will absorb more electrons and
will appear darker in the final pictures. Density differences are due to
differences in the contour of the coated surfaces of the specimen. The
image produced will be in three dimensions, and the pictures are called
scanning electron micrograph (SEM).

xxv

BIOLOGY 1
DB014

DB014 Lab Manual

EXPERIMENT 1: BASIC TECHNIQUES IN MICROSCOPY

Course Learning Objective: Conduct basic biology laboratory work on
microscopy, tissues, genetics information, pollen germination and plant
growth by applying manipulative skills.
(P3, CLO 2, PLO 2, MQF LOD 2)

Learning Outcomes:
At the end of this lesson, students should be able to:
i. Handle microscope properly
ii. Obtain accurate images.
iii. Calculate the actual magnification.

Student Learning Time (SLT):

Face-to-face Non face-to-face

2 hours 0

Experiment 1.1: Handling Microscope Properly

Before doing the following experiments, you must read and understand
the basic techniques of using a microscope. Refer to Introduction to
Microscopy.

Apparatus

Compound microscope

Experiment 1.2: To obtain accurate images 1

Apparatus

Compound microscope

Materials
‘e’ prepared slide
Cross threads prepared slide (3 colours)
Transparent ruler (10 mm size)
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DB014 Lab Manual

Procedures and Observation

Images under the microscope

1. Observe the ‘e’, cross threads and transparent ruler prepared slide
using the 4x objective lens.

2. What do you observe using the 4x objective lens? Draw what you
have observed.

3. What do you observe using the 10x objective lens? Draw what you
have observed.

Experiment 1.3: Magnification

Procedures and Observation

1. Determine the actual magnification of a specimen by using the
formula below.

Magnification power x Magnification
Actual magnification = of objective lens a power
g of ocular lens

2. Calculate the actual magnification in Table 1.1. n
i

Table 1.1 Actual magnification of a sfpecimen
i

Actual magnification c
a

Magnification Magnification power of obtjective lens
power of ocular i

lens 4x 10x 40xo 100x
n

10x p
o
w
e
r

Updated: 20/05/2020 o
f2

o
b
j
e
c

DB014 Lab Manual

Questions

Part A: Handling microscope properly.

1. A microscope is called a compound microscope when it consists
of more than one set of …………………………………

2. Condenser and iris diaphragm are useful to coordinate
…………………………………………...

3. Based on laboratory practices, what do you use to clean the
microscope lenses?

4. While observing a moving microorganism under a microscope,
you found that the organism has moved out of the field of view
to the right. In order to keep observing the microorganism, which
direction do you move your slide (right/left)?

5. How do you adjust the slide when the specimen is out of the field
of view to the top?

Part B: To obtain accurate image

1. Which is the shortest objective lens?
2. Which objective lens should you use when you begin to focus a

specimen?
3. Which objective lens should be in position before you store a

microscope?
4. Which objective lens will still remain in focus when placed at the

longest working distance from the specimen?
5. When using an ocular lens with 10x magnification power, which

objective lens should be used to obtain the following actual
magnification?
(a) 100 times of its diameter
(b) 1000 times of its diameter

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DB014 Lab Manual

EXPERIMENT 2: ANIMAL TISSUES

Course Learning Objective: Conduct basic biology laboratory work on
microscopy, tissues, genetics information, pollen germination and plant
growth by applying manipulative skills.
(P3, CLO 2, PLO 2, MQF LOD 2)

Learning Outcomes:
At the end of this lesson, students should be able to:
i. Identify the animal tissues under the light compound

microscope.
ii. Identify the structure of epithelial tissue, muscle tissue and

connective tissue

Student Learning Time (SLT):

Face-to-face Non face-to-face

2 hours 0

Introduction 4

Four main types of animal tissues are epithelial tissue, muscle tissue,
connective tissue and nerve tissue.

Epithelial tissues form the surface lining of other tissues either inside
or outside the body. Epithelial tissues consist of epithelial cells which
are closely arranged between the cells. Epithelial tissues have
specialized functions for growth, protection, excretion and absorption.

The main function of muscle tissues is for contraction. Based on
structures, muscle tissues can be divided into three types: striated
muscle, smooth muscle and cardiac muscle. Physiologically, muscle
tissues can be divided into voluntary muscle and involuntary muscle.

Bone tissue, cartilage tissue and blood tissue are grouped into
connective tissues. The functions of the connective tissues are for
support, connecting tissues and organ, transport and defence.
Connective tissues consist of three components: cell, ground substance
(substance which is deposited in between the connective tissues and

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DB014 Lab Manual

serves to hold the cells together) and fibre. The fibre together with the
connective tissue is embedded in the matrix.

The main function of nerve cells or neuron is to conduct nerve
impulses. Apart from neuron, the nerve tissue consists of neuroglia
which gives internal support.

Apparatus

Compound microscope

Materials

Prepared slides of epithelial tissues
a) Simple squamous
b) Simple cuboidal
c) Simple columnar

Prepared slides of nerve tissue
a) Motor neuron

Prepared slides of muscle tissues
a) Smooth

Prepared slides of connective tissue
a) Blood

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DB014 Lab Manual

Procedures and Observation

1. Examine the slides given using either 10x or 40x objective lens.
2. Draw and label the structure and organization of each of the

following tissues. Use the figures provided to assist you in your
cell investigation.

(i) Epithelial tissues
a) Simple squamous epithelium
b) Simple cuboidal epithelium
c) Simple columnar epithelium

(ii) Nerve tissue
a) Motor neuron

(iii) Muscle tissues
a) Smooth

(iv) Connective tissues
a) Blood

Figure 2.1 Simple squamous epithelium 6
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DB014 Lab Manual

Figure 2.2 Simple cuboidal epithelium

Figure 2.3 Simple columnar epithelium

erythrocytes

white blood cell

Figure 2.4 The types of blood cells 7
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Figure 2.5 Longitudinal section of a smooth muscle

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DB014 Lab Manual

Figure 2.6 Motor neuron 9
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DB014 Lab Manual

Questions:

Describe the types, functions and distributions of tissues below:

a) Epithelial tissues
- Simple squamous epithelium
- Simple cuboidal epithelium
- Simple columnar epithelium

b) Nerve tissue
- Motor neuron

c) Muscle tissues
- Smooth

d) Connective tissues
- Blood

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DB014 Lab Manual

EXPERIMENT 3: PLANT TISSUES

Course Learning Objective: Conduct basic biology laboratory work on
microscopy, tissues, genetics information, pollen germination and plant
growth by applying manipulative skills.
(P3, CLO 2, PLO 2, MQF LOD 2)

Learning Outcomes:
At the end of this lesson, students should be able to:
i. Identify the plant tissue under the light compound microscope
ii. Identify different types of ground and vascular tissue.

Student Learning Time (SLT):

Face-to-face Non face-to-face

2 hours 0

Introduction

Permanent tissues consist of mature cells that have specialized
structure and functions. Permanent tissues can be divided into three:
dermal tissues (epidermal and peridermal), ground tissues
(parenchyma, sclerenchyma, collenchyma and endodermis) and
vascular tissues (xylem and phloem).

Apparatus

Compound microscope

Materials

Prepared slides of cross sections of monocot/dicot stem and root

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DB014 Lab Manual

Experiment 3.1: Structures of plant tissues in plants

1. Examine the prepared slides of any types of plant tissues
(root/ stem)

2. Identify the distribution of the tissues (parenchyma,
collenchyma, sclerenchyma, phloem and xylem).

3. Draw the arrangement of the tissues. Note the differences in the
following characteristics: cell size, shape, wall thickness and
stained parts.

4. Select a section and observe it under 40x objective lens. Draw
and label the section showing the different tissues (parenchyma,
collenchyma, sclerenchyma, phloem and xylem). Use the figures
provided to assist you in your tissue investigation.

Figure 3.1 Structure and distribution of vascular bundles
incross section of monocot root. (Adapted from
www.sci.waikato.ac.nz)

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DB014 Lab Manual
Figure 3.2 Cross section of monocot root

Figure 3.3 Structure and distribution of vascular bundles in
cross section of dicot root

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DB014 Lab Manual

Figure 3.4 Cross section of dicot root.
(Adapted from www.sci.waikato.ac.nz)

Figure 3.5 Structure and distribution of vascular bundles
in cross section of monocot stem.

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DB014 Lab Manual

Figure 3.6 Structure and distribution of vascular bundles in
cross section of dicot stem.
(Adapted from www.bio.miami.edu)

Monocot Dicot

Figure 3.7 Structure of different plant tissues in cross
section of monocot and dicot stem

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DB014 Lab Manual

Questions:

1. Describe the types, functions and distributions of tissues below:

a) Ground Tissues
- Parenchyma cells
- Collenchyma cells
- Sclerenchyma cells

b) Vascular Tissues
- Xylem tissue
- Phloem tissue

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DB014 Lab Manual

EXPERIMENT 4: CELL DIVISION - MITOSIS

Course Learning Objective: Conduct basic biology laboratory work on
microscopy, tissues, genetics information, pollen germination and plant growth by
applying manipulative skills.
(P3, CLO 2, PLO 2, MQF LOD 2)

Learning Outcomes:
At the end of this lesson, students should be able to identify different stages
of mitosis.

Student Learning Time (SLT):

Face-to-face Non face-to-face

2 hours 0

Introduction

In most tissues, new cells are formed as a result of mitosis. If the
chromosomes of such cells are selectively stained with a dye such as
aceto-orcein, stages in mitosis can be observed. An example of a tissue that
undergoes mitosis is the meristematic tissue. This tissue is located in the cell
division zone of the apical meristem at the root tip.

Apparatus

Compound microscope
Prepared slides of various stages of mitosis

Procedures and Observation

Experiment 4.1: Prepared slides of mitosis

1. Examine the prepared slides of various stages of mitosis.
2. Draw and label the stages of mitosis observed.

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DB014 Lab Manual

Questions:
1. Describe the four stages of mitosis.
2. Where does mitosis actively take place in plants?

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DB014 Lab Manual

EXPERIMENT 5: POLLEN GERMINATION

Course Learning Objective: Conduct basic biology laboratory work on
microscopy, tissues, genetics information, pollen germination and plant growth by
applying manipulative skills.
(P3, CLO 2, PLO 2, MQF LOD 2)

Learning Outcomes:
At the end of this lesson, students should be able to:
i. Identify the various shapes and structures of pollen grains.
ii. Explain the reproductive structure of flowering plant.

Student Learning Time (SLT):

Face-to-face Non face-to-face

2 hours 0

Introduction

The growth of a pollen tube is a fascinating phenomenon. Pollen grains are
morphologically simple, small structures which contain two nuclei when
released from the anther at anthesis. When a viable pollen grain lands on the
stigma of a compatible flower, it produces a tube several hundred to several
thousand micrometers long in which the pollen nuclei travel to the ovary of
the flower. The process of tube formation is a relatively uncomplicated
example of growth and development. Pollen germination represents a short,
yet very critical event in a series of steps leading to the double fertilization of
the ovule.

Experiment 5.1: Pollen germination

Apparatus

Compound microscope
Cavity well slides and cover slips
Dropper
Dissecting microscope
Dissecting needle
Scalpel/ Razor blade/ lancet

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DB014 Lab Manual

Material
Hibiscus and Spider Lily flower

Procedures and Observations

1. Add 1-3 drops of water on a cavity well slide and transfer pollen
grains onto it.

2. Cover with a cover slip.
3. Observe the pollen grains under a compound microscope using 40x

objective lens.
4. Draw and label your observation.
5. Remove the sepals and petals from hibiscus flower by gently pulling

them off the receptacle.
6. Locate the stamen, each of which consists of a thin filament with a

pollen-filled anther of the tips. Note the number of stamen.
7. Locate the pistil. The stigma at the top of the pistil is often sticky.

The style is a long, narrow structure that leads from the stigma to the
ovary.
8. Observe the stamen and pistils of the hibiscus flower with dissecting
microscope.
9. Draw and label the male and female reproductive system of hibiscus
flower.

Questions:

1. Compare the hibiscus and spider lily pollen grain structure.
2. Why stigma at the top of the pistil is often sticky?
3. Why pollen is important in fertilization?

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DB014 Lab Manual

EXPERIMENT 6: PLANT GROWTH

Course Learning Objective: Conduct basic biology laboratory work on
microscopy, tissues, genetics information, pollen germination and plant growth by
applying manipulative skills.
(P3, CLO 2, PLO 2, MQF LOD 2)

Learning Outcomes:
At the end of this lesson, students should be able to:
i. Observe and record the growth of a bean sprout.
ii. Measure growth parameter (height of plant, length of root and length of

leaf).

Student Learning Time (SLT):

Face-to-face Non face-to-face

2 hours 0

Introduction

Growth is a fundamental characteristic of all living organism. Growth is
always associated with an increase in size. However, increase in size is not
the sole factor to define growth. There are stages in organism development
where there is an increase in cell numbers but not body size. Growth and
development goes together in tandem. Development can also be described as
an increase in complexity. For metazoan, the growth can be divided into
three phases; cell division, cell enlargement, and cell differentiation. Growth
can be measured by plotting parameters such as length, height, mass, surface
area, volume and number against time. Sigmoid curve is a typical growth
pattern for many organisms.

Apparatus

Cotton wool/ filter paper
Graph paper
Plastic container (eg. Mineral bottle, plastic cup etc.)
Ruler
Thread

Materials

Mung beans 20
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DB014 Lab Manual

Procedures and Observation

1. Place the cotton wool / filter paper into the plastic container.
2. Pour adequate water into the container.
3. Place the 15 mung bean seeds on the cotton wool.
4. Observe what happens to the mung beans for 8 days. Water them

daily.
5. The observation includes the length of root, the height of plant, and

the length of the leaf.
6. Record your observation in Table 6.1.

Table 6.1 Mean length of root, the height of plant, and the length of

the leaf

Days 0 1 2 3 4 5 67

Mean length of

root/ hypocotyls

(mm)

Mean height of

plant (mm)

Mean length of

leaves (mm)

7. Plot the growth curves of the bean sprouts in a single graph.

Questions:

1. Based on the graph drawn, what is the shape of the growth curve?
Explain why.

2. Which of the parameter shows the most rapid growth? Explain your
answer.

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BIOLOGY 2
DB024