MATRICULATION DIVISION BIOLOGY · 2020. 7. 18. · 1.3 Biology 1 Course Learning Outcome At the end...

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

Malaysia National Library

Biology Laboratory Manual

Semester I & II

DB014 & DB024

Fourth Edition

eISBN 978-983-2604-47-1

iii

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.

iv

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


v

CONTENTS

Page

Foreword iv

Content v

Learning Outcomes vii

Introduction

x

SEMESTER I

Experiment Title

1 Basic Techniques In Microscopy

1

2 Animal Tissues

4

3 Plant Tissues

11

4 Cell Division – Mitosis

17

5 Pollen Germination 18

6 Plant Growth 20

vi

SEMESTER II

Experiment Title

7 Diversity of Bacteria

22

8 Plant Diversity: Gymnosperm

24

9 Animal Diversity: Invertebrates and Vertebrates

27

10 Chloroplast in Aquatic Plant 36

11 Homeostasis

39

12 Coordination

45

References 53

Acknowledgements 54

vii

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)

viii

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)

ix

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.

x

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.

xi

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.

xii

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.

xiii

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

xiv

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

xv

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).

xvi

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

xvii

Table 1 Components of microscope and their functions

Component Function

1a. Ocular lens or eyepiece lens:

This lens is found at the top of the

microscope. It normally has a

magnification power of 10x or 15x.

1b. Ocular control knob.

1a. Magnifies the real image and

converts it to a virtual image to be

viewed by user’s eyes.

1b. To adjust and compensate the

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:

You will hear clicking sounds when the

objective lens is in its correct position

above the specimen. You may practise

this by rotating and changing the

objective lenses.

The structure to which the objective

lenses are mounted. By gently rotating

the nosepiece, you may choose the

objective lens you want and correctly

place it over the specimen.

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):

Coarse specimen field = 5 mm

b. Low-power objective lens (10x):

Small specimen field = 2 mm

c. High-power objective lens (40x):

Very small specimen field = 0.5 mm

d. Oil immersion objective lens (100x):

Immersion oil is placed in the space

a. Used to scan the specimen before

identifying the specific part to be

viewed further.

b. Used to view major part of the

specimen.

c. Used to view specific part of the

specimen.

xviii

between the specimen and objective lens

to reduce the light refraction from the

specimen.

Very small specimen field = 0.2 mm

d. Used to view microorganisms such

as bacteria and microstructures in the

cells.

5a. Stage:

The horizontal surface which has a hole

in the centre to allow light from below

to focus on the specimen.

5b. Slide clips.

5c. Slide adjustment knob

a. The horizontal surface on which a

specimen is placed.

b. The stage is usually equipped with

slide clips to hold the slide in place.

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:

Condenser is located immediately under

the stage

6b. Condenser control knob.

6c. Condenser lens knob.

a. Used to focus and deliver light to the

specimen.

b. Used to adjust the condenser.

c. Used to focus the light.

7. Iris diaphragm:

An adjustable light barrier of iris type

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.

Controls the amount of light entering

and leaving the condenser.

8a. Off/on switch.

8b. Light control knob.

Please ensure that either of the switches

is OFF or MINIMUM, respectively

before you use the microscope.

The source of light is a tungsten bulb

located at the base of a microscope.

xix

9. Body arm

The metal part used to carry a

microscope.

10. Base

The heavy cast metal part used as the

base and for support.

11. Coarse adjustment knob:

Use this knob only when using low-

power objective lens. Rotate this knob

carefully and observe what happens.

Does the stage or the body tube move?

Used to bring specimen into focus by

moving the stage to the specimen.

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.

xx

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.

xxi

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,

xxii

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.

xxiii

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.

xxiv

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

xxv

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).

BIOLOGY 1

DB014

DB014 Lab Manual

Updated: 20/05/2020 1

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

Apparatus

Compound microscope

Materials

‘e’ prepared slide

Cross threads prepared slide (3 colours)

Transparent ruler (10 mm size)

DB014 Lab Manual

Updated: 20/05/2020 2

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


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

formula below.

Actual magnification =

2. Calculate the actual magnification in Table 1.1.

Table 1.1 Actual magnification of a specimen

Actual magnification

Magnification

power of ocular

lens

Magnification power of objective lens

4x 10x 40x 100x

10x

Magnification power

of objective lens

Magnification

power

of ocular lens

x

a

g

n

i

f

i

c

a

t

i

o

n

p

o

w

e

r

o

f

o

b

j

e

c

DB014 Lab Manual

Updated: 20/05/2020 3

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

DB014 Lab Manual

Updated: 20/05/2020 4

EXPERIMENT 2: ANIMAL TISSUES





Learning Outcomes:


i. Identify the animal tissues under the light compound

microscope.

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

connective tissue



2 hours 0

Introduction

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

DB014 Lab Manual

Updated: 20/05/2020 5

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

DB014 Lab Manual

Updated: 20/05/2020 6


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

DB014 Lab Manual

Updated: 20/05/2020 7

Figure 2.2 Simple cuboidal epithelium

Figure 2.3 Simple columnar epithelium

Figure 2.4 The types of blood cells

erythrocytes

white blood cell

DB014 Lab Manual

Updated: 20/05/2020 8

Figure 2.5 Longitudinal section of a smooth muscle

DB014 Lab Manual

Updated: 20/05/2020 9

Figure 2.6 Motor neuron

DB014 Lab Manual

Updated: 20/05/2020 10

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

DB014 Lab Manual

Updated: 20/05/2020 11

EXPERIMENT 3: PLANT TISSUES





Learning Outcomes:


i. Identify the plant tissue under the light compound microscope

ii. Identify different types of ground and vascular tissue.



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

DB014 Lab Manual

Updated: 20/05/2020 12

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)

DB014 Lab Manual

Updated: 20/05/2020 13

Figure 3.2 Cross section of monocot root

Figure 3.3 Structure and distribution of vascular bundles in

cross section of dicot root

DB014 Lab Manual

Updated: 20/05/2020 14

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.

DB014 Lab Manual

Updated: 20/05/2020 15

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

http://www.bio.miami.edu/

DB014 Lab Manual

Updated: 20/05/2020 16

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

DB014 Lab Manual

Updated: 20/05/2020 17

EXPERIMENT 4: CELL DIVISION - MITOSIS


microscopy, tissues, genetics information, pollen germination and plant growth by

applying manipulative skills.


Learning Outcomes:

At the end of this lesson, students should be able to identify different stages

of mitosis.



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


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.

DB014 Lab Manual

Updated: 20/05/2020 18

Questions:

1. Describe the four stages of mitosis.

2. Where does mitosis actively take place in plants?

DB014 Lab Manual

Updated: 20/05/2020 18

EXPERIMENT 5: POLLEN GERMINATION





Learning Outcomes:


i. Identify the various shapes and structures of pollen grains.

ii. Explain the reproductive structure of flowering plant.



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

DB014 Lab Manual

Updated: 20/05/2020 19

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?

DB014 Lab Manual

Updated: 20/05/2020 20

EXPERIMENT 6: PLANT GROWTH





Learning Outcomes:


i. Observe and record the growth of a bean sprout.

ii. Measure growth parameter (height of plant, length of root and length of

leaf).



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

DB014 Lab Manual

Updated: 20/05/2020 21


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 6 7

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.

BIOLOGY 2

DB024

DB024 Lab Manual

Updated: 20/05/2020 22

EXPERIMENT 7: DIVERSITY OF BACTERIA


diversity of living things, chloroplast in aquatic plant, homeostasis and

coordination by applying manipulative skills.


Learning Outcomes:


i. Identify different shapes of bacteria.

ii. Apply the use of oil immersion with high magnification

(oil immersion lens)



2 hours 0

Introduction

Bacteria can be identified based on shapes, position of flagella and Gram

staining. The three most common shapes are spheres, rods and spirals

Apparatus

Compound microscopes with 100x objectives lens

Materials

Prepared slides of different types of bacteria

Immersion oil

Lens tissue papers

Methylated spirit (only for specific use)

DB024 Lab Manual

Updated: 20/05/2020 23


1. Observe the prepared slide under the microscope.

(Caution: Use immersion oil only for 100x objective lens).

2. Draw your observation.

(Caution: Draw only the bacteria and not artifacts such as air

bubbles, dust, fibre, etc.)

REMINDER: Make sure the oil immersion lens is properly

cleaned at the end of experiment (refer to section

INTRODUCTION TO MICROSCOPY)

Questions:

1. Describe various shapes of bacteria.

2. What are the precautions taken when handling immersion oil?

DB024 Lab Manual

Updated: 20/05/2020 24

EXPERIMENT 8: PLANT DIVERSITY - GYMNOSPERMS





Learning Outcomes:

At the end of this lesson, students should be able to identify morphological

reproductive structure of gymnosperm.



2 hours 0

Introduction

Gymnosperms

Gymnosperms are plants that produce naked seeds on scaly structure, named

strobilus or cones. Its sporophyte generation produces 2 types of spores:

microspores that will develop into male gametophytes and megaspores that

develop into female gametophytes which is nutritionally dependent on living

sporophyte. The four main phyla of gymnosperms are Cycadophyta,

Ginkgophyta, Coniferophyta and Gnetophyta.

Experiment 8.1: Diversity of Gymnosperm

Apparatus

Compound microscope

Dissecting microscope

Razor blade

Tile

DB024 Lab Manual

Updated: 20/05/2020 25

Materials

Fresh specimens:

Cycas sp. (Phylum Cycadophyta)

Male and female cones of Cycas sp.

Gnetum sp. (Phylum Gnetophyta)

Male and female cones of Gnetum sp.

Pinus sp. (Phylum Coniferophyta)

Male and female cones of Pinus sp.

Prepared slides:

Megaspores of Cycas sp. cones

Microspores of Cycas sp. cones

Megaspores of Gnetum sp. cones

Microspores of Gnetum sp. cones

Megaspores of Pinus sp. cones

Microspores of Pinus sp. cones


1. Observe the given specimens.

2. Draw and label the specimens to show their morphological

differences.

3. Examine the prepared slides of megaspores and microspores. Draw

and label the megaporophyll, megasporangium, microsporophyll and

microsporangium.

DB024 Lab Manual

Updated: 20/05/2020 26

Questions:

1. State the unique characteristics of gymnosperms.

2. Cycadophyta is primitive gymnosperms that evolved from ferns.

Which characteristic of the fern is still maintained in the Cycadophyta?

3. In Pinus sp., male cone matured earlier than female cone. Describe

how this species ensures the success of pollination?

4. Gnetophyta is considered as a linkage between gymnosperms and

angiosperms based on a few characteristics found in Gnetophyta. State

the characteristics.

DB024 Lab Manual

Updated: 20/05/2020 27

EXPERIMENT 9: ANIMAL DIVERSITY – INVERTEBRATES AND

VERTEBRATES





Learning Outcomes:

At the end of this lesson, students should be able to identify unique

characteristics of invertebrates and vertebrates.



2 hours 0

Introduction

Animal classification is a matter of sorting out their similarities and

differences, and then placing them into groups. Among the most basic criteria

by which animals are sorted is whether or not they possess a backbone

(vertebral column). More than 90% of animal species found in the world

belonged to invertebrates.

Invertebrates

Invertebrates are multicellular animals without vertebral column. Many

invertebrates such as jellyfish or worm have a fluid-filled hydrostatic skeleton

that function as a support mechanism. Other invertebrates such as insects and

crustaceans possess hard outer shell for protection. There are different phyla

of invertebrates. The most common phyla of invertebrates include the

Annelida, Arthropoda, Mollusca and Echinodermata.

Vertebrates

Vertebrates are classified in the phylum Chordata. Members of the subphylum

Vertebrata comprise of animal with backbones and spinal cord.

Approximately 64,000 species of vertebrates have currently been described.

Vertebrata is the largest subphylum of Chordata, and contains many familiar

DB024 Lab Manual

Updated: 20/05/2020 28

groups of large land animals. Vertebrates are the animals from the groups of

jawless fishes, bony fishes, sharks and rays, amphibians, reptiles, mammals,

and birds. All vertebrates have the following characteristics during the

development stage:

(i) notochord

(ii) dorsal tubular nerve cord

(iii) pharyngeal slits present in early embryonic development

(iv) post anal tail

The above characteristics may undergo changes or diminish during the

embryonic development.

Experiment 9.1: Invertebrates

Materials

Preserved or live specimens:

Phylum Cnidaria: - Hydra (Obelia sp.)

Phylum Platyhelminthes: - Planaria (Dugesia sp.)

Phylum Nematoda: - Roundworm (Ascaris lumbricoides)

Phylum Annelida: - Earthworm (Pheretima sp.)

Phylum Mollusca: - Garden snail (Achatina sp.)

Phylum Arthropoda: - Horseshoe crab (Tachypleus sp.)

- Grasshopper (Valanga sp.)

- Giant Freshwater Prawn (Macrobrachium sp.)

- Millipede (Julus sp.)

Phylum Echinodermata: - Star fish (Asterias sp.)

Procedures and observation

1. Observe the morphological characteristics of the given specimens.

2. Draw and label the unique characteristics of your specimens.

3. Refer to Appendix 9.1 to assist your investigation.

DB024 Lab Manual

Updated: 20/05/2020 29

Questions:

1. State the unique characteristics of invertebrates.

2. Name the two forms of polymorphism in cnidarian.

3. List some adaptations possessed by Platyhelminthes as endoparasites.

4. What is meant by metameric segmentation in annelids and state its

advantages.

5. What is the function of clitellum in oligochaetes?

6. What is the function of radula in molluscs?

7. What is the function of the water vascular system in a star fish?

DB024 Lab Manual

Updated: 20/05/2020 30

Experiment 9.2: Vertebrates

Materials

Preserved or live specimens:

Class Chondrichthyes (cartilaginous fishes): - Stingray (Raja sp.)

Class Osteichthyes (bony fishes): - Scad (Selar sp.)

Class Amphibia: - Frog (Rana sp.)

Class Reptilia: - Snake (Boa sp.)

Class Aves: - Pigeon (Columba sp.)

Class Mammalia: - Rat (Rattus sp.)


1. You are given specimens from different classes within the subphylum

Vertebrata.

2. Observe the morphological characteristics of the given specimens.

3. List the characteristics of each specimen.

4. Refer to Appendix 9.2 to assist your investigation.

Questions:

1. State the unique characteristics of vertebrates.

2. List the common orders in Mammalia and give an example in each

order.

3. Compare the morphological characteristics of a shark and scad.

4. List the adaptations that enable amphibians to adapt to terrestrial life.

5. Compare the morphological characteristics of Mammalia and Aves.

DB024 Lab Manual

Updated: 20/05/2020 31

Appendix 9.1: Characteristics of selected phyla in invertebrates

1. Phylum Porifera

Example: Leucosolenia sp. (Sponge)

Characteristics:

Multicellular animal; simple body structure; asymmetry body; the cells

are arranged loosely without real tissue; the body consists of two

layers of cells-pinacoderm on the external surface and choanoderm on

the internal; the choanoderm consists of flagellated collar cells, folded

body walls and numerous pores; the body walls are supported by a

skeleton from calcium, spicules from silica or spongin fibers; the body

cavity is called spongocoel; there is a large body opening, osculum, at

one end; undifferentiated nervous system.

2. Phylum Cnidaria

Example: Obelia sp. (Hydra), Aurelia sp. (jellyfish)

Characteristics:

Lower stage multicellular animal; radial symmetry; diploblastic (two

germ layers); nematocyst; nervous system consists of network of nerve

cells, tentacles around the manubrium (mouth), alternation of

generation between polyp and medusa stage.

3. Phylum Platyhelminthes

Example: Dugesia sp. (planarian), Taenia sp. (tapeworm)

Characteristics:

Flatworm; Most members are parasitic except class Turbellaria.

Parasitic representative lacks both respiratory and circulatory systems,

simple digestive system or none at all; excretory system is called the

‘flame’ cell; the mouth on the ventral side; the eyes on the dorsal side.

4. Phylum Nematoda

Example: Ascaris sp. (roundworm), Brugia sp. (roundworm)

Characteristics :

Non-segmented, pseudocoelomates. Complete digestive tract. The

muscles of nematode are all longitudinal. Body covered with tough

cuticles.

DB024 Lab Manual

Updated: 20/05/2020 32

5. Phylum Annelida

Example: Pheretima sp. (earthworm), Nereis sp. (ragworm)

Characteristics:

Segmented body; the segments are normally separated by

compartments which are called septum; the chaetae on the body are

used to help in locomotion.

6. Phylum Mollusca

Example: Achatina sp. (garden snail), Anadara sp. (cockle)

Characteristics:

Has a large muscular body, visceral organ present; has radula; a mantle

whose function is to secrete shell.

7. Phylum Arthropoda

Characteristics:

Segmented animals; mostly terrestrial; exoskeleton from chitin; paired

jointed appendages; tripoblastic and bilateral symmetry; respiratory

system consists of the tracheal system which opens through spiracles;

aquatic Arthropods respire through external gills; the sensory and

nervous systems are concentrated at the head region.

Example:

(a) Class Crustacea - crab

(b) Class Chilopoda - centipede

(c) Class Arachnida - spider

(d) Class Merostomata - horseshoe crab

(e) Class Insecta - grasshopper

(f) Class Diplopoda - millipede

8. Phylum Echinodermata

Example: Asterias sp. (star fish)

Characteristics:

Triploblastic; locomotion using tube feet; skeleton from calcium

carbonate plates; simple digestive system; part of the coelom is

modified to become water vascular system.

DB024 Lab Manual

Updated: 20/05/2020 33

Appendix 9.2: Characteristics of selected classes in vertebrates

1. Class: Chondrichthyes (cartilaginous fishes)

Example: shark, ray fish

Characteristics:

Flexible endoskeletons made of cartilage rather than bone; clasper –

slender, grooved section present in mature male; caudal fin is

heterocercal; sharp teeth evolved from the jagged scales that cover the

abrasive skin; sensory pore in front of the mouth; non-movable

rudimentary eye lids: a pair of spiracles; gill slits; cloaca and lateral line

system.

2. Class: Osteichthyes (bony fishes)

Example: scad, gold fish, tilapia

Characteristics:

Bony skeleton with many vertebrae; operculum – a lateral bony flap;

caudal fin is homocercal; teeth in jaws; unpaired median fins; paired

pelvic and pectoral fins.

3. Class: Amphibia

Examples: Bufo sp.(toad), Rana sp. (frog)

Characteristics:

Non-scaly, soft and moist glandular skin which also functions as a

respiratory surface, tetrapods, thin webbed feet for paddling and

crawling, double nostrils which open up into the mouth cavity and lead

to the lungs; wide mouth with small teeth; closed circulatory system, the

heart is divided into three chambers.

4. Class: Reptilia

Example: snake, lizard, tortoise, crocodile

Characteristics:

Hard, dry, horny scales shed periodically; 2 pairs of legs with five

fingers; lungs for gaseous exchange; three-chambered heart (crocodile –

four-chambered heart).

DB024 Lab Manual

Updated: 20/05/2020 34

5. Class: Aves

Example: pigeon, eagle, duck

Characteristics:

Moved on two feet; forelimbs, with three clawed fingers which can be

adapted to form wings; body and wings covered with feathers;

adjustable hind limbs for walking, swimming or perching; keratin beaks,

with great variety of shapes suitable for different diets; and lay eggs.

Adaptations to fly:

(a) Wings

(b) Hollow bones which are strong and light

(c) Effective respiratory and circulatory systems provide the cells

with enough oxygen to permit a high metabolic rate for

tremendous muscular activity to fly

(d) Complete digestive system

(e) Well-developed nervous system

Other characteristics:

(a) legs covered with scales

(b) no teeth

(c) four-chambered heart

(d) efficient lungs with air sacs

(e) no urinary bladder

6. Class: Mammalia

Example: human, rat, cat, whale, platypus

Major characteristics:

Hair which insulates and protects the body; mammary glands, produces

milk for young; possess three small bones (malleus, incus, stapes) in the

middle ear.

Other characteristics:

(a) Skull with two occipital condyle

(b) Upper and lower jaws with differentiated teeth

(c) Outer ear lobes and movable eye lids

(d) Four limbs for terrestrial adaptation

(e) Lungs for breathing and the presence of voice box

DB024 Lab Manual

Updated: 20/05/2020 35

(f) Four chambered heart

(g) Exothermic

(h) The presence of urinary bladder

(i) Internal fertilization, where the fertilized egg developed in the

female uterus. The embryonic membrane consists of amnion,

chorion and allantois.

DB024 Lab Manual

Updated: 20/05/2020 36

EXPERIMENT 10: CHLOROPLAST IN AQUATIC PLANT





Learning Outcomes:


i. Prepare the slide of an Elodea leaf.

ii. Examine the internal structure of an Elodea leaf: the cell membrane,

cell wall and chloroplasts



2 hours 0

Introduction.

Elodea is an excellent plant for our studies of photosynthesis and cells

because the leaves are only a few cells thick so they will be easy for us to

observe under the microscope to look at cells and cell parts.

Figure 10.1 Elodea sp.

DB024 Lab Manual

Updated: 20/05/2020 37

Figure 10.2 Cells of Elodea sp.

Figure 10.2 shows leaf cells of an Elodea sp. These living plant cells are

viewed by light microscopy.

With a microscope we can see the chloroplasts move around inside living

cells. Chloroplasts and the other living parts of the cells are found near the

cell wall, but not usually in the center of the cell. Since they contain

chlorophyll, which are green, chloroplasts can be seen without staining and

are clearly visible within living plant cells. However, viewing the internal

structure of a chloroplast requires the magnification of an electron

microscope.

Apparatus

Microscope

Slide

cover slip

Material

A sprig of Elodea sp.

DB024 Lab Manual

Updated: 20/05/2020 38


1. Obtain an Elodea sp. leaf from the main plant, and place it on a clean

slide.

2. Place one drop of fresh water onto the leaf, and carefully place the cover

slip on top of the leaf.

3. Place the slide under the microscope and observe under low power and

medium power.

4. Turn the microscope to high power and draw what you see. Label the

chloroplasts (the green objects), the cell wall and the cell membrane.

Questions:

1. Why would we use an aquatic plant (Elodea sp) for this experiment

instead of a terrestrial plant?

2. Where do aquatic plant get the CO2 for photosynthesis?

3. What organelle in plants responsible for photosynthesis ?

4. What is the pigment found in this organelle that absorbs light to power

photosynthesis?

5. If a plant were under water and was photosynthesizing, what gas would

be visibly bubbling from the plant?

6. Why do chloroplast move in Elodea?

DB024 Lab Manual

Updated: 20/05/2020 39

EXPERIMENT 11: HOMEOSTASIS





Learning Outcomes:

At the end of this lesson, students should be able to detect the effects of

exercise on the body temperature and pulse rate in a human body system.



2 hours 0

Introduction

Homeostasis is to maintain constant internal environment of the living

organism through physiological regulation. Various organs are responsible

for regulating different environmental variables. In human, heart rate and

temperature need to be regulated within certain set ranges. Hypothalamus for

example plays important roles in regulating body temperature through

negative feedback mechanism. When body temperature increases,

thermoreceptors will detect this changes and cause physiological adjustment

to bring the temperature back to normal range. Medulla oblongata likewise

plays the similar functions to regulate body pulses.

Apparatus

Clinical thermometer

Digital pulse monitor

Height scale

Stop watch

Weighing scale

Precaution to students: Do not attempt this activity if you have a health

problem or if you are recovering from an illness.

DB024 Lab Manual

Updated: 20/05/2020 40


1. Class will be divided into groups for conducting this exercise. Before

the exercise, students who undertake this exercise should fill vital

parameters in Table 11.1. Use Appendix 11.1 to assist you in

calculating of body mass index (BMI).

Table 11.1 Vital parameters taken before exercise for each student

Student’s

Name Gender

Height

(m)

Body

weight

(kg)

BMI

Average

initial

pulse rate

Average

initial body

temperature

2. Sit down comfortably on a chair and take 5 minutes to settle, locate

your pulse, and count the initial number of pulses per minute. Take

three readings and record the average in Table 11.1.

3. Measure your initial body temperature. Take three readings and record

the average in Table 11.1.

4. For the first exercise, each student will walk around in class for 5

minutes. Immediately measure the average pulse rate and average body

temperature and record in Table 11.2.

5. For the second exercise, each student will run on the spot for 10

minutes (intermediate intensity). Immediately measure the average

pulse rate and average body temperature and record in Table 11.2.

6. While resting, take the average pulse rate and average body

temperature for every 10 minutes interval in half an hour (three times)

and record in Table 11.2.

7. Analyse on class basis the changes in pulse rate and temperature after

different exercises according to the gender and BMI and record them in

Tables 11.3 and 11.4.

DB024 Lab Manual

Updated: 20/05/2020 41

Table 11.2 Vital parameters taken after exercises for each student

Pulse rate

(beats/minute)

Body temperature

(0C)

Initial

(From Table 11.1)

After walking

(first exercise)

After running

(second exercise)

Recovery

period

After 10

minutes

After 20

minutes

After 30

minutes

DB024 Lab Manual

Updated: 20/05/2020 42

Table 11.3 Results of the effect of exercises on the pulse rate

(for the whole class)

Gender BMI No. of

students

Pulse rate (beats/minute)

Initial

After

walking

(first

exercise)

After

running

(second

exercise)

Recovery period

After 10

minutes

After 20

minutes

After 30

minutes

Male

< 18.5

18.5 – 24.9

25 – 29.9

> 30

Female

< 18.5

18.5 – 24.9

25 – 29.9

> 30

DB024 Lab Manual

Updated: 20/05/2020 43

Table 11.4 Results of the effect of exercises on the body temperature

(for the whole class)

Gender BMI No. of

students

Body temperature (OC)

Initial

After

walking

(first

exercise)

After

running

(second

exercise)

Recovery period

After 10

minutes After 20

minutes After 30

minutes

Male

< 18.5

18.5 – 24.9

25 – 29.9

> 30

Female

< 18.5

18.5 – 24.9

25 – 29.9

> 30

DB024 Lab Manual

Updated: 20/05/2020 44

Questions

1. What is the significance of homeostasis in human body?

2. What are the normal ranges of human body temperature and pulse rate?

3. Name two other examples of vital parameters in human that are

regulated by negative feedback mechanism.

Appendix 11.1

BMI Categories:

• Underweight = < 18.5

• Normal weight = 18.5 – 24.9

• Overweight = 25 – 29.9

• Obesity = > 30

Source: Adapted from WHO, 1995; WHO, 2000 and WHO 2004.

Body weight (kg)

(Height)2 (m) Body mass index (BMI) =

DB024 Lab Manual

Updated: 20/05/2020 45

EXPERIMENT 12: COORDINATION





Learning Outcomes:


i. Identify the receptor of taste buds in tongue.

ii. Display reflex action of nervous system.



2 hours 0

Introduction

Living organisms depend on the information obtained from the environment

for their survival. Most of the information about the environment detected by

the special sense organs called the receptor organ. These receptors receive

stimuli such as light, sound, temperature, taste and smell in the environment.

Although stimuli and sensory organs are different, the sequences of events

that occur in the receiving of stimuli are the same (see diagram). External

stimuli are received by the sense organs such as ears, eyes, nose, skin and

tongue. From sensory organs, the external stimuli are received by one or

more sensory neurons that become impulses. Sensory neurons have many of

the same character, but different from one organ to another organ that is

sensitive to a specific stimulus. Example: there are sensory neuron sensitive

to light and sensory neurons that are sensitive to vibration. Central nervous

system will translate these impulses and the information obtained is used by

the organism to react the stimulus.

Stimulus → Receptor organ → Neuron → Impulses transmit → CNS

DB024 Lab Manual

Updated: 20/05/2020 46

In human there are two types of reflex, which are:tendon reflex and organ

reflex. Tendon reflex consists of patella, achilles andplantar reflex. Reflex

organs consist of pupil accommodation, ciliospinal and swallow reflex. All

the reflexes are to coordinate our body protection.

Experiment 12.1: Effector and Receptor

All experiments must be done in pairs.

Apparatus

100 ml beaker/ paper cup

Cotton bud

Petri dish

Materials

Bitter gourd extract

Lemon juice

Salt solution

Sucrose solution

Warm water


1. Rinse mouth thoroughly with tap water.

2. Dip cotton bud into sucrose solution and ask your partner to touch on

the areas of the tongue shown in Figure 12.1.

DB024 Lab Manual

Updated: 20/05/2020 47

a – tip

b – side-front (left and right)

c – middle-front

d – middle-back

e – side-back (left and right)

f – below-middle-front

Figure 12.1: Sensitive parts of a tongue

3. Redraw Figure 12.1 and indicate the following indicators beside

labels a to f.

Too sweet - +++

Sweet - ++

Slightly sweet - +

Not sweet - 0

4. Repeat procedures 1 to 3 with the following solutions:

a) Salt water

b) Lemon juice

c) Bitter gourd extract* (10%- 10g in 90 mL water)

* Chop and mash the bitter gourd.

5. Record the results for each solution:

Too salty/ sour/ bitter - +++

Salty / sour/ bitter - ++

Slightly salty/ sour/ bitter - +

Not salty/ sour/ bitter - 0

DB024 Lab Manual

Updated: 20/05/2020 48

Results

Yourself

TASTE TASTE AREA

a b c d e f

Sweet

Salty

Sour

Bitter

Your Partner

TASTE TASTE AREA

a b c d e f

Sweet

Salty

Sour

Bitter

Questions

1. State whether same or different receptor give different taste stimulus.

Explain your answer.

DB024 Lab Manual

Updated: 20/05/2020 49

Experiment 12.2: Reflex

Apparatus

Torch light

Wooden Ruler/Reflex hammer


A. Tendon Reflex

i) Patella reflex

1. Sit on the table and let your legs freely hanging and

dangling over the edge of the table.

2. Close your eyes, your partner will knock on your knee

tendon by using wooden ruler from the side of the leg.

3. Observe knee jerking.

Figure 12.2A Tendon Reflex

DB024 Lab Manual

Updated: 20/05/2020 50

ii) Achilles reflex

1. Ask your partner to stand on the knee down on one

knee on a stool.

2. One of the legs should be bent downward stretching

the gastrocnemius tendon.

3. Your partner surprisingly knocks on your

gastrocnemius tendon using wooden ruler from the

side of the leg.

4. Observe the other foot jerking (Figure 12.2B)

5. Record your observations in Table 12.2A.

Figure 12.2B Achilles reflex

DB024 Lab Manual

Updated: 20/05/2020 51

Table 12.2A: Observation of tendon reflex

Types of reflex Observation

Patella

Achilles

B. Organ Reflex.

i) Pupil accommodation reflex

1. Ask your partner to focus an object at a distance.

2. Observe your partner’s pupil.

3. Ask your partner to focus on a very close object.

4. Observe your partner’s pupil.

5. Flash your partner’s eyes starting from the side of the

eyes using a torch light.

6. Observe your partner’s pupil (Figure 12.2C)

7. Record your observations in Table 12.2B.

Figure 12.2C Pupil accommodation reflex when expose to light

DB024 Lab Manual

Updated: 20/05/2020 52

Table 12.2B Observation of organ reflex

Types of reflex Observation

Pupil

i) Distance object

______________________________________

ii) Close object

______________________________________

iii) Exposure to light

a) With flashlight

______________________________________

b) Without flashlight

_______________________________________

Questions:

1. What are the functions of reflex?

2. Explain the flow of reflexion process.

3. List the types of neurons that are involved in this experiment.

53

REFERENCES

Campbell, N. A., Reece, J. B., Urry, L. A., Cain, M. L., Wassermen, S. A.,

Minorsky, P. V. & Jackson, R. B. (2018). Biology. (11th Ed.). Pearson

Benjamin Cummings. USA.

Lawrence, E. (2016). Henderson’s Dictionary of Biological Terms (16th Ed.),

Prentice Hall.

Solomon, E. P., Berg, L. R. & Martin, D. W. (2018). Biology. (11th Ed.).

Nelson Education, Ltd, Canada.

Morgan J. G & Carter M. E. B & Stout (2015). Investigating Biology:

Laboratory Manual (8rd Edition), Pearson Education Limited.

www.bio.miami.edu

www.crochetspot.com

www.k-state.edu

www.math.arizona.edu

www.news.makemeheal.com

www.pc.maricopa.edu

www.quia.com

www.sci.waikato.ac.nz

www.sfsu.edu

www.sharewhy.com

www.sols.unlv.edu

www.stolaf.edu

www.users.rowan.edu

www.vcbio.science.ru.nl

www.wikispace.psu.edu

54

AKNOWLEDGEMENT

The Matriculation Division wishes to extend heartfelt thanks and utmost

gratitude to the following individuals who have actively participated in

marking the review a success. We are grateful for the support and guidance

provided by those involved, namely:

Dr. Hajah Rosnarizah binti Abdul Halim Director of Matriculation Division,

Ministry of Education

Dr. Shah Jahan bin Assanarkutty Deputy Director, Academic Sector,


Mohd Junaidi bin Abd Aziz Senior Chief Assistant Director,

Academic Sector,


Mdm. Salbiah binti Mohd. Som Selangor Matriculation College

Mr. Abdul Aziz bin Abdul Kadir Malacca Matriculation College

Mdm. Nizaha binti Zulkifli Negeri Sembilan Matriculation College

Mdm. Lena Maizura binti Basaruddin Perak Matriculation College

Mdm. Rudziah binti Umar Selangor Matriculation College

Mdm. Rohaiza binti Rozali Assistant Director, Academic Sector,


Mr. Ruslan bin Achok Assistant Director, Academic Sector,


MATRICULATION DIVISION BIOLOGY · 2020. 7. 18. · 1.3 Biology 1 Course Learning Outcome At the end...

Documents

Transcript of MATRICULATION DIVISION BIOLOGY · 2020. 7. 18. · 1.3 Biology 1 Course Learning Outcome At the end...