ANSWER SCHEME

EXPERIMENT 1: BASIC TECHNIQUES IN MICROSCOPY

1. Title BASIC TECHNIQUES IN MICROSCOPY

2. Objectives i. To obtain accurate images
ii. To determine the depth of field
iii. To determine the field of view
iv. To calculate the actual magnification
v. To apply the use of oil immersion with high magnification (oil
immersion lens)

3. Apparatus Compound light microscope

4. Materials ‘e’ prepared slide
Cross threads prepared slide (3 colours eg: yellow, red and blue)
Transparent ruler (10 mm size) or graph paper prepared slide
Prepared slide of bacteria
Lens tissue papers
Immersion oil
Methylated spirit (only for specific use)
5. Introduction Microscope can be used to magnify an object, determining the size of an
object and observing fine details of an object.

In compound microscopes, the actual magnification is calculated as the

magnification of objective lens multiplied by the magnification power of
the ocular lens.

The image that seen under microscopes was inverted because it goes
through two lens systems, and because of the reflection of light rays. The
two lenses it goes through are the ocular lens and the objective lens.

The depth of field is a measure of the thickness of a plane of focus. As the

magnification increases, the depth of field decreases. At low
magnification we might be able to see the entire volume of a specimen,
but when increase the magnification we may only be able to see one
surface of the specimen.

The field of view is inversely proportional to the magnification of the

objective lens. The specimen appears larger with a higher magnification
because a smaller area of the object is spread out to cover the field of
view of eye.

Microscopes magnify an object's appearance by bending light. Higher

magnification means the light is bent more. At a certain point, the light is
bent so much that it can't make it through the objective lens. At that
point – usually around 100x for standard lab microscopes we need to put
a drop of oil between the specimen and the objective lens. The oil
"unbends" the light to stretch out the working distance and make it
possible to image at high magnifications.
6. Procedures 1.1.1 Images under the microscope
 1. ‘e’ prepared slide was observed using 4x objective lens and the
observation was draw in the paper.
2. The position of ‘e’ was determined.

1.1.2 The depth of field

1. The position of the thread on the slide was observed with naked
eyes and the color of thread were identified.
2. The cross threads were observed under the microscope using 4x
and 10x objective lens.
3. The depth of field was determined when the power of objective
lens increases.

1.1.3 The field of view

1. A transparent ruler was placed on the stage.
2. The transparent ruler was observed using the 4x, 10x and 40x
objective lenses.
3. The observation was draw in the lab report.

Exercise 1.3: Oil Immersion Objective Lens

1. The prepared slide of bacteria was observed under the
microscope.
2. The observation was draw.

7. Result Exercise 1.1: Images, Depth of Field and Field of View of the Microscope

i. Image under the microscope

● Drawing of letter ‘e’

No. Criteria Marks

1. Drawing ‘e’ letter must be inverted 1
2. Title of the drawing and state the actual 1
magnification (40x)
Total Marks 2

Actual magnification power = 40x

Title: prepared slide of letter ‘e’

ii. Depth of Fields
● Draw table for location of thread
Position of Naked eyes Under
thread microscope : 4x
and 10x

Top Position of thread under the

Middle microscope and naked eye
Bottom should be the same

The depth of field decreases when the power of objectives lens increase
= [1 marks]

iii. Field of View

Drawing of transparent ruler at 40x actual magnification

No. Criteria Marks

1. Drawing of transparent ruler 1
2. Title of the drawing and state the actual 1
magnification (40x)
Total marks 2

Answer the question on page 4: Procedure 3 and 4

No. Procedure Marks
3. The diameter of field of view for the 4x 1
objective lens is 3.5 mm or 3.5 x 103 µm.
The diameter of field of view for the 10x
objective lens is 1.5 mm or 1.5 x 103 µm. 1

The diameter of field of view for the 40x 1

objective lens is 0.375 mm or 3.75 x 102
µm.

*calculation (for lecturer references):

* 1mm = 1 x 103 µm
DOF at low magnification = High mag
power
DOF at high magnification Low mag
power

1.5mm = 40x10
x 10x10

x = 0.375 mm

4. Cork seen under 40x objective lens

= 20 cell horizontal x 10 cell vertical
 Under 40x objective lens
Horizontally,
20 cell = 3.75x102µm
1 cell = 3.75x102µm/ 20 cell
1 cell = 0.19 x 102 µm

Vertically, 1
10 cell = 3.75x102µm
1 cell = 3.75x102µm/10 cell
1cell = 0.38 x102µm

Size of a cork cell 1

= 0.19 x 102 µm x 0.38 x102µm
= 7.22 µm2

1
Total marks 6

Exercise 1.2: Magnification

● Complete Table 1.1 on page 3
Actual Magnification
Magnification Magnification power of objective
power of lens
ocular lens 4x 10x 40x 100x

10x 40x 100x 400x 1000x

Exercise 1.3: Oil immersion objective lens

● Drawing of prepared slide using oil immersion objective lenses

No. Criteria Marks
1. Drawing seen under microscope 1
2. Title of drawing and state the actual 1
magnification (1000x)
Total marks 2

Actual magnification: 1000 x

 Prepared slide for Gram stain – Bacillus subtilis

Actual magnification: 1000 x

Prepared slide for Gram stain – Neisseria subflava

8. Discussion ● Power of objectives lens

● The lowest objective lens is 4x objective lens which is use to scan
the whole image of the specimen
● Next, for objective lens 10x and 40x which is use to focus specific
section of image of the specimen
● 100x objective lens also known as oil immersion objective lens
which is use to observe the specimen at the highest resolution
and usually use to observe microorganism such as bacteria and
parazoa

● Describe image seen under the microscope

- Larger
- Inverted
- Virtual

● State the relationship between depth of field, magnification power

and image seen under the microscope
- As the magnification power increase,
- the depth of field is smaller/ narrower
- the less image can be focused vertically

● State the relationship between field of view, magnification power and

image seen under the microscope
- As the magnification power increase,
- the field of view is smaller/ narrower
- the less image can be focused

● State the formula for calculating the magnification power.

 Actual magnification = magnification power of objective lens x
magnification power of ocular lens
9.
Conclusions 1. Images seen under the microscope are bigger than the objects/
clear/ inverted.
2. The higher the magnification power the smaller/ narrower field of
view and depth of field
3. By using the formula given, we are able to calculate the actual
magnification
4. The prepared slide of bacteria can be observed by using oil
immersion objective lens (100x).

10. References At least THREE references written according to APA system:

Author. (Year). Title of Book/journal. Publisher. Country/place where it’s

published.

Excluding lecture notes and manual book.

11. Answers on Part A
Questions No. Answer Marks
1. A 1
2. A 1
3. A 1
4. A 1
5. D 1
6. A 1
7. B 1
D 1

Part B.
1. Lens tissue //alcohol 70% // xylene // diethyl 1
ether
2. Left 1
3. By moving the slide downward 1

Part C
1. Ocular lens and objective lens 2
2. Light intensity 1

Total Marks=14 marks

Maximum marks= 10 marks
Total
EXPERIMENT 2: PLANT TISSUES

NO. MANIPULATIVE SKILLS MARKS

1. Ability to follow instruction / teamwork 5
- Use light compound microscope according to safety rule.
2. Ability to handle apparatus 5
- Broken apparatus: 0%
- Able to conduct experiment following the correct procedure.
3. Ability to handle microscope 5
- Able to display specimen clearly under low and high magnification
- Able to focus and observe plant tissues under light compound
microscope.
4. Cleanliness of workplace 5
- No messy table
Total 20

NO. CONTENT MARKS

1. Title 1
2. Objectives 1
3. Apparatus 1
4. Materials 1
5. Procedures 2
6. Introduction 8
 This section introduces theory, background and motivation in the
context of the experiment.
7. Result / Observation 9 x 5 = 45
Drawing must have:
Drawing and labelling 1) Title
Under 4x objective lens: 2) Scientific
- Monocot Stem drawing
- Monocot Root 3) Actual
- Dicot Stem Magnification
- Dicot Root 4) Labelling
(minimum 2)
Under 40x objective lens: Max: 5 marks each
- Epidermis diagram
- Parenchyma
- Collenchyma
- Sclerenchyma
- Xylem, Phloem and Cambium

8. Discussion and Questions 15

Question 1
Monocot Stem Dicot Stem 1
1(a) Vascular bundle Vascular bundle 1
scattered arranged in ring form
No cambium Cambium exist between
phloem and xylem 1

Monocot Root Dicot Root 1

1(b) Obvious endodermis Xylem and phloem
and pericycle surrounded by
endodermis
Xylem and phloem is Xylem is arranged like a
arranged alternately star shape
Any 2

Question 2
(Based on structure)
Parenchyma
Any 2
● Primary cell walls that are thin and flexible
● Lack of secondary cell walls
● Large central vacuoles
● Living cell with protoplast
● Loosely arranged between cells

Any 2
Collenchyma
● Thicker primary wall but unevenly thickened walls
● The thickening at the corners of the cell
● Chemical composition in cell wall: cellulose,
hemicellulose and pectin (no lignin)
● Living cells with protoplast
Any 2
 ● Arrangement of cells are more closely packed / with little
or no intercellular space
● Cell more elongated

Sclerenchyma Any 2
● Walls are evenly thickened
● Lignin rich in the cell wall
● Very hard
● Impermeable to water due to presence of lignin 1
● Arrangement of cell is very closely packed

(Based on function)
Parenchyma
● Cell division before matured
● Store of mineral include starch grain, oil droplets, water
and salts in dicotyledone
● Photosynthesis (some cells contain chloroplast)
● Support
● Gas exchange (has intercellular space from the loosely
packed of cell)

Collenchyma
● Support
● Becomes meristemetic & produces cork cambium in
dicotyledon stem that undergoes secondary thickening

Sclerenchyma
● Support and strenghten plant tissues

9. Conclusion 3
1. List down the types of plant tissues
2. Monocot and dicot stem and root can be distinguished
based on the distributions of tissues.

10. References 3

i.At least THREE references written according to APA system:

Author. (Year). Title of Book/journal. Publisher. Country/place

where it’s published.

Excluding lecture notes and manual book.

Total 80
EXPERIMENT 3: TRANSPORT ACROSS MEMBRANES

NO. MANIPULATIVE SKILLS MARKS

1. Ability to follow instruction / teamwork 10
- preparing potato strips & sucrose solutions.
2. Ability to handle apparatus 10
- Broken apparatus: 0%
- Able to conduct experiment following the correct procedure.
3. Cleanliness of workplace 5
- No messy table
Total 25

NO. CONTENT MARKS

1. Title 1
2. Objectives 1
3. Apparatus 1
4. Materials 1
5. Procedures 3
6. Introduction 10

This section introduces cells as a selectively permeable membrane. (pg 14 manual

lab)

7. Result 25

Exercise 3.1: Determination of molarities of sucrose solution

Molarity 0.1M 0.2 M 0.3 M 0.4 M 0.5 M 2x5=10m

Volume of
1.0 M 2 4 6 8 10
sucrose (ml) 5m
 Volume of 18 16 14 12 10
distilled
water (ml) (2m) (2m) (2m) (2m) (2m)

Students should show a calculation how to get the result in the table by using 5m
M1V1 = M2V2
eg :1.0M x V1 = 0.1M x 20ml
V1 = 0.1M x 20ml/1.0M
= 2ml
volume distilled water=total volume solution - V1
= 20ml - 2 ml = 18ml

Draw a standard graph of water potential against molarities of sucrose solution.

Osmotic potential,atm 5m

(1m)
(1m) - Tittle of graph (1m)
- Scale (1m)

Molarity mol dm-3 (1m)

- drawings with correct graph, scale and labelling

Draw the graph of the change in weight of potato strips against the molarities of
the sucrose solutions.

Weight of potato strip,gm (1m)

- Title of graph (1m)
- Scale (1m)

(1m)

Molarity, mol dm-3 (1m)

- drawings with correct graph, scale and labelling

8. Discussion 25

• Compare experimental results with each other

• Explain errors

Students manually kept on touching the potatoes with hands which could
have defected the sugar distribution across the potatoes surface and altering
the osmosis results.
 More than one potato was required in the experiment to get the 15 potato
strips, thus us having several different sucrose sources and by the fact that
each potato has different levels of nutrition, the results might have a
deficiency inaccuracy in this area.

• Comment on trends shown by a graph

Graph change in weight of potato strips against the molarities of the sucrose
solutions are inversely proportional which positive value of change in weight
potato strips at 0.1-0.3M of sucrose solution, zero value change of weight
potato strips at 0.3-0.4M and negative value change of of weight potatostrips
at 0.4-0.5M

• Explain the relationship between changes in weight of potato strips

and the molarities of sucrose solution (includes the direction of water
movement and condition of cell)

The sucrose solution from 0.1 M to 0.3 M are hypotonic to potato cell sap.
Water diffuses into the cells by osmosis and the potato strips become turgid
and longer.

The sucrose solution from 0.4M to 0.5M are hypertonic to potato cell sap.
Water diffuses out of the cells by osmosis and the potato strips become
flaccid and shorter as the concentration sucrose solution are increases.

The concentration sucrose solution is isotonic from 0.3M to 0.4M (obtained

from the graph) at which no change in weight of potato strips. This is because
isotonic solution where no net water movement into and out of the cells

9. Conclusion 5
State whether the aim of the experiment has been achieved or not, summarise the
key features of the method used, and summarise the most important results.

10. References 3
i.At least THREE references written according to APA system:

Author. (Year). Title of Book/journal. Publisher. Country/place where it’s

published.

Excluding lecture notes and manual book.

Total 75
EXPERIMENT 4: CELL DIVISION – MITOSIS

NO. MANIPULATIVE SKILLS MARKS

1. Ability to follow safety rule 4
- Use light compound microscope according to safety rule.
- Able to keep the workplace clean.
2. Ability to find and display specimen under the microscope 4
- Able to display specimen clearly under low and high magnification
3. Ability to handle experiment 4
- Ability to prepare slide of onion root tip.
- Ability to use the correct materials and apparatus.
4. Ability to Focus 4
- Able to focus specimen under light compound microscope.

5. Ability to report result 4

- Able to draw the four stages of mitosis
Total 20

NO. CONTENT MARKS

1. Title 1
2. Objectives 1
3. Apparatus 1
4. Materials 1
5. Procedures 3
6. Introduction 10

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 and shoot apex.

7. Result Total: 15
 Diagram: 1
Title: 1
Label: 1
Magnification:
1

MAX: 3 marks

Diagram: 1
Title: 1
Label: 1
Magnification:
1

Total: 4 marks
Prophase
Actual magnification: 400x

Diagram: 1
Title: 1
Label: 1
Magnification:
1

Total: 4 marks
 Diagram: 1
Title: 1
Label: 1
Magnification:
1

Total: 4 marks

Metaphase
Actual magnification: 400x

Anaphase
Actual magnification: 400x
 Telophase
Actual magnification: 400x

8. Discussion 20

(Exercise 4.1: Prepared slides of mitosis) 10

E.g.;
● All four stages of mitosis can be observed from the prepared slides.
10
(Exercise 4.2: Preparation of onion root tips slide)
E.g.;
● During prophase , chromosomes become shorten and thicken.
● During metaphase, chromosomes arrange on equatorial plate /
metaphase plate.
● During anaphase, chromosomes as a sister chromatids split and
move to opposite poles of the cells.
● During telophase, chromosomes arrive at the opposite poles and
become decondensed.
● Metaphase was the most frequent stage observed since it is the
longest stage in a cell cycle.
● Sometimes, the stages of mitosis cannot be observed because the
region cut is not the root tip area.
 ● For better observation, eraser can be used to press the root tips in
order to spread out the cell.
● The four stages of mitosis cannot be clearly observing due to the
limitation by the quality of microscope used in this experiment.

9. Conclusion 5

An onion root tips slide was prepared. The four stages of mitosis which
were prophase, metaphase, anaphase and telophase were identified and
drawn. The metaphase stage was more frequently observed compared to
other stages.

10. References 3

i.At least THREE references written according to APA system:

Author. (Year). Title of Book/journal. Publisher. Country/place where it’s

published.

● http//: www.nature/stages of mitosis.com

Excluding lecture notes and manual book.

11. Answers on Questions 20
3
1. Why is the root tip placed in acetic alcohol?
As a fixative. The function of acetic alcohol is to stabilise the structures
and prevent chemical and structural changes whilst the material is 3
undergoing staining and mounting.

2. What is the purpose of using HCl in this experiment? 2

HCl is used to destroy the substances that unite the cells (usually
pectin), but it does not destroy the cell walls. The hydrochloric acid also 10
has the ability to stop the process of mitosis so that its phases can be
observe without possibilities of the cells moving into another stages.

3. What is the stage in the mitosis that is frequently observed? Why?

Metaphase. Because the stage is the longest. 2

4. Describe the chromosome behavior at each stage in mitosis.

Prophase – Chromosomes condense become shorten and thicken.
Metaphase – Chromosomes arrange on equatorial plate / metaphase
plate.
Anaphase – Chromosomes as a sister chromatids split and move to
opposite poles of the cells.
Telophase – Chromosomes arrive at the opposite poles and become
decondensed.

5. Where does mitosis actively take place in plants?

Root tips and shoot apex.

Total 80
EXPERIMENT 5: INHERITANCE

NO. MANIPULATIVE SKILLS MARKS

1. Ability to follow instruction / teamwork 5
- Technique of blood sampling
2. Ability to handle apparatus 10
- Broken apparatus: 0%
- Able to conduct experiment following the correct procedure.
3. Cleanliness of workplace 5
- No messy table
Total 20

NO. CONTENT MARKS

1. Title 1
2. Objectives 1
3. Apparatus 1
4. Materials 1
5. Procedures 3
6. Introduction 10

This section introduces theory, background and motivation in the context of the
experiment.
7. Result 15

- Determination of own genotype 2

- Calculation of the observed and expected frequencies of every genotype
in the class.

Observed frequency: observed / total students 5

Expected frequency: Mendel’s ratio x total students 5
*at least ONE calculation is shown

- Determination own blood group 1

- Complete the table of possible genotypes and its frequency 2

8. Discussion 20

Interpret the results and explain the significance of finding.

Interpretation you have to:
• compare your results with accepted values
• compare your experimental results with each other
• explain errors
• comment on the suitability of the method used in the experiment.
 Discussing results should need to point out the trends and explain why they are as
they are. Ask whether
● The results agree with the model/theory? Why not?
● What experimental limitations were there that might have affected the
accuracy of your data?
● Were results obtain expected?

Precautions taken while handling the experiment

● Discuss the safety measures should be taken and why
● Is there any other safety requirement needed to be considered?

Marks are given accordingly to the ability to interpret and discuss about the result
as well as the other aspects such as precautionary action.

9. Conclusions 5

State type of inheritance controlled by single gene. 1

State the highest and the lowest frequency of phenotype in the class. 2
State the highest and the lowest percentage of blood group in the class. 2

10. References 3

v.At least THREE references written according to APA system:

Author. (Year). Title of Book/journal. Publisher. Country/place where it’s
published.

Excluding lecture notes and manual book.

11. Answers on Questions 20

5.1

1. Carrier means that individuals inherited heterozygous genotype contains 2

recessive alleles BUT hidden by dominant allele.

2. If left-handed was recessive, that student inherited recessive allele from 2

both his parents.

3. Expected frequency is 3/16. Tongue rolling = 3/4, Attached earlobe = 1/4. 2

4. Expected frequency is (1/4)6 . 2

5.2

1. To allow more blood flowing to the fingertip. 1

2. To avoid blood contamination from the others in order to prevent 1

infectious diseases.

3. The first drop of blood is diluted with the alcohol wipe that was put over 2
the tip of your finger. Therefore, to get an accurate test result, the first drop needs
to be discarded as to get fresh blood.

2
4. A different toothpick prevents the contents of slide 1 from crossing to
slide 2, which could invalidate. Avoiding contamination from one slide to the
other. 3

5. Yes. Blood group O is the universal donor. It can be donated to people

with type A, B or AB and O. This is due to the absence of antigen A and B on the
red blood cells of people with blood type O. Therefore, when a person having type
A, B or AB blood receives type O blood, there wouldn't be any agglutination
reaction between the antibody A present in type A blood (commonly known as
anti-A) or antibody B present in type B blood (anti-B) or antibody A and B
present in type AB blood with the blood cells from the person with type O blood.

6. Each biological parent donates one of their two ABO alleles to their
child. A mother who is blood type O can only pass an O allele to her daughter. 3
A father who is blood type AB could pass either an A or a B allele to his daughter.
This couple could have children of either blood type A (O from mother and A
form father) or blood type B (O from mother and B from father). Having a
daughter with blood type O is impossible. Thus , in this case it is clear that the
babies were swapped in the nursery.
Total 80
EXPERIMENT 6: BASIC TECHNIQUES IN ISOLATING DNA

NO. MANIPULATIVE SKILLS MARKS

1. Ability to follow instruction / teamwork 10
2. Ability to handle apparatus 10
- Broken apparatus: 0%
- Able to conduct experiment following the correct procedure.
3. Cleanliness of workplace 10
- No messy table
Total 30

NO. CONTENT MARKS

1. Title 1
2. Objectives 1
3. Apparatus 1
4. Materials 1
5. Procedures 3
6. Introduction 10

Each chromosome is a single thread-like structure made up of long molecules of

DNA combined with histone protein. The DNA molecule is made up of many
sections called genes. Shortly before cell divisions occurs, each DNA molecules
replicates itself. So one thread of chromosome becomes two identical chromatids.
As the two chromatid are identical, they will have identical genes. These identical
genes are known as allele. In this experiment, you will rupture fruit cells, thus
releasing their content such as proteins, DNA, RNA, lipids, ribosomes and various
small molecules. DNA is then suspended by alcohol as supernatant layer.

The purity of DNA will require further steps. After the isolation of nucleic acids,
the solution is still contaminated with proteins which can be removed. To check
the success of the removal, a purity determination is performed, which is based on
the different absorption characteristics of the proteins and the nucleic acids using
UV spectrophotometer.
7. RESULT 15
Draw and label THREE layers of solution in the boiling tube.
- Top layer: Cold alcohol
- Middle layer: supernatant containing DNA filament
- Bottom layer: Sieved liquid
8. Discussion 20
- on structure and importance of DNA
DNA thread found in supernatant layer because the molecular weight of DNA is
lighter than other molecules/organelles.

The DNA that can be observed in this experiment is not pure DNA (using naked
eyes.)

Structure of DNA
- DNA building block is a nucleotide: pentose sugar deoxyribose, a
phosphate and nitrogenous bases
- Nucleotide are linked by covalent bonds to form an alternating
sugar-phosphate backbone in one polynucleotide
- The 3’ carbon of one sugar is bonded to the 5’ phosphate of adjacent
sugar to form a phosphodiester bond.
- DNA molecule consisting of two polynucleotide chains arranged in a
coiled double helix
- The two polynucleotide chains must run in opposite direction that are said
to be antiparallel to each other
Importance of DNA
- Carrying the hereditary material
9. Conclusions 5
- DNA of the fruit isolated in the form of DNA filament 1
- DNA filament found suspended in the supernatant layer between alcohol 2
and sieved liquid 2
- DNA filament isolated from the fruit’s cells content such as proteins,
RNA, lipids, ribosomes and various molecules
10. References 3

v.At least THREE references written according to APA system:

Author. (Year). Title of Book/journal. Publisher. Country/place where it’s

published.

Excluding lecture notes and manual book.

11. Answers of Questions 10

1. (a) To emulsify fat

To create a polar environment to dissolve DNA
(b) To precipitate DNA
(c) To use the heat:
- to soften the tissue and allow the medium/solution to penetrate
- to denature enzyme and protein
2. To lysed (break down) the fruit cell wall

Total 70