AS201

BACHELOR OF SCIENCE (HONS) BIOLOGY

CELL BIOLOGY BIO 411
LABORATORY REPORT 1
MADAM SARINA BINTI HASHIM

NAME NO MATRIC CLASS

MUHAMMAD IFFAT HANIF BIN MOHD RUDIN 2022899874 AS2011B

NUR FARRISYA BINTI MOHAMAD ZIN 2022478792 AS2011B

NURUL AIHAN BINTI AHMAD HILMI 2022697432 AS2011B

SHARIFAH NUR ALYA INSYIRAH BINTI SYED ADLAN 2022697594 AS2011B

OBJECTIVES

1. To become familiar with compound and dissecting microscopes, their parts and
functions.
2. To acquire the skill of calibrating prior to measuring field of view and cell size using
the ocular and stage micrometer.
3. To examine prepared slides of connective tissues.

INTRODUCTION

All cells' physical maintenance is managed by the extracellular matrix (ECM). However,
the concept that the ECM has a passive role to play in cellular activity has been refuted. It is now
acknowledged to partake in several biological activities, such as cell division, proliferation, and
migration. The ECM is a prominent component of connective tissues, where it has been compared
to the "glue" that holds cells together. It is the non-cellular component of tissues. While the
structural and essential building block of life is the cell. Cell biology is the study of cells, including
their fundamental makeup and organelle functions. Cells were initially found by a biologist, Robert
Hooke.
The objectives of this experiment, "Exploring and Viewing: Cell and ECM," are to
familiarize ourselves with compound and dissecting microscopes, as well as their components
and uses. Second, learn how to calibrate an ocular and stage micrometer before measuring the
field of view and cell size, and third, look at connective tissue preparation slides. A compound
light microscope can be used to magnify the sixth prepared slides, and a dissecting microscope
can be used to see other specimens including leaves, pond water, and insects.

MATERIALS
1. Compound and dissecting microscope
2. Specimens:
● Leaf
● Pond water
● Insects
● Prepared slides
3. Others:
● Immersion oil
 ● Stage and ocular micrometer
● Distilled water
● Labels
● Glass slides and cover slides
● Tissue papers
● Forceps
● Razor blades
● Dropper bottles
● Pasteur pipettes
● Petri dishes

4. Prepared slides of
● Elastic tissue
● Lily anther mature pollen
● Skin thin human
● Human blood smear
● Mammal intestines injected sections
● Hyaline cartilage
● Compact bone
● Chick embryo 27-29 hr OR chick embryo 20-22 hr

PROCEDURES

Task 1: Basic Microscopy

1. The dissecting and compound microscope were carefully obtained from the display
cabinets.
2. The revolving nosepiece was turned so that the lowest power objective lens is “clicked”
into position.
3. The slide was placed on the stage and fasten it with the stage clips.
4. The objective lens and the stage from the side were looked at and the coarse focus knob
was turned so that the objective lens moves downward.
5. The eyepiece was looked at and the illuminator and diagrams were adjusted for the
greatest amount of light.
 6. The coarse adjustment was slowly turned so that the objective lens goes up. The
adjustment continued until the image came into focus.
7. The microscope slide was moved around so that the image is the center of the field of
view and the mirror, illuminator or diaphragm were readjusted for the clearest image.
8. After that, we were able to change to the next objective with only minimal use for the
focusing adjustment. Steps 4 through 7 were repeated with the highest power objective
lens in place.
9. The glass part of the lenses could not be touched with the fingers. Only special lens paper
was used to clean the lenses(Kim wipes).
10. When finished, the tube was raised, the low power lens was clicked into position and the
slide was removed.

Task 2: Calibration for sample measurement

1. With the stage micrometer in focus, the eyepiece was rotated until the ocular micrometer
was aligned to the stage micrometer.
2. The scale was scanned along it, and the point was found where the ocular micrometer
scale was directly aligned to the scale of the stage micrometer.

RESULT

TASK 1
COMPOUND MICROSCOPE
Part Name Function

A Body Tube To reflect light up to the viewer's eye

B Nose Piece Allows for quick change of objectives

C Objective lens Usually there are three or four objective lenses found in
microscope which are 10x, 40x, and 100x power

D Stage Clips Used to keep the slide in place

E Diaphragm Used to vary the amount of light passing through

the slide

F Light Source Sends light up through the diaphragm and through

t the slide for viewing

G Eye Piece Lens that presents at the top to see objects under study

H Arm Used to safely transport microscope

I Stage The flat platform that used to place slides for

. observation

J Coarse Adjustment Knob Used to make large changes in focus

K Fine Adjustment Knob Used for small adjustments of focus

L Base Provide basal support for microscope

M Stage control Move the slide

Table 1
 Comparison and contrast between the functions of a compound microscope and dissecting
microscope

Compound Microscope Dissecting Microscope

● Widely used in research industry

● Made of several lenses that are put together
● Optical microscopes where they use visible light

To view very small objects that could not be seen To inspect larger objects such as minerals and
with naked eyes insects

Use transmitted light illumination (light is passed Use incident light illumination (light is reflected
through the sample) off the surface of the object)

Have much higher optical resolution with Have lower optical resolution power where the
magnification ranging from about 40x to 1000x magnification typically ranges between 6x to
50x

Provides two-dimensional image Provides three-dimensional image

Have a single optical path Have a different optical path

Table 2

TASK 2

40 div = 10 μm (4x magnification)

100 div = 10 μm
1 div = 100/10
= 0.1 μm
 14
14 div = x 10 μm
40

= 3.5 μm

26
26 div = x 10 μm
100
= 2.6 μm

40
40 div = x 1 μm
40

= 1 μm
 TASK 3

Compact Bone cell

Compact bone
400x total magnification
Cell size: 17.25 μm

Calculation of the cell size:

69
= x 10 μm
40

= 17.25 μm
DISCUSSION

TASK 1

A microscope is a machine that generates enlarged images of small objects. The

microscope also allows an extremely close look at minute structures at a scale fitting for
examination and analysis. A microscope's magnifying power is a dimensionless ratio that
expresses the number of times the object being examined appears to be enlarged. Compound
microscopes are commonly used for laboratories. It has thirteen parts and its function. All their
parts and functions have been classified in table 1. While in table 2 , it is shown the differences
and similarity between compound microscope and dissecting compound.The standard objective
lenses that were fixed on turret microscope are 4x, 10x, 40, and 100x. The 4x, 10x, 40x and 100x
are collectively known as dry lenses. The 100x objective is known as the oil immersion lens where
it requires the use of immersion oil. There are some rules that need to be followed when using an
oil immersion lens. Some of them never use an oil immersion lens without the oil and never get
oil on any other lenses. Oil immersion is very crucial to eliminate any air gaps and light loss due
to refraction which is the bending of light as light passes through the slide to air to the objective
lens. When using a microscope, it is important to follow all the procedure. When focusing the high-
power objective lens, never use the coarse adjustment because it can break the slide or damage
the lens. When finished, either raise or lower the tube or stage, and after that click the low power
lens position and remove the slide. The total magnification of an image is calculated by multiplying
the magnification of the objective lens by the magnification of the ocular lens. For example,
magnification of an objective lens is 40x and magnification of the ocular lens is 10x. So, the total
magnification will be 400x magnification.

TASK 2

The eyepiece graticule is an important part of the eyepiece that allows for measurements.
It's a disc made of glass marked with a scale from 0 to 100 (μm). Depending on the type of
eyepiece, the eyepiece graticule may be fitted onto the eyepiece or be purchased separately and
attached onto the eyepiece for measurements. During calibration, it's important that each
individual lens be calibrated separately starting with the lowest power objective. This makes it
possible to have distinct calibration power for each.
 During the calibration process, the stage micrometer has to be aligned with the eyepiece
graticule scale followed by taking the readings. Readings obtained from the scales are then used
for the purposes of calculating the calibration factor. The process involves aligning the zero
point/lines on both scales first. Once the two points have been aligned properly, then the one
calibrating has to scan through the scale lines to find a point where the lines of both scales align.
It's important to ensure that the starting point of the two scales align properly before scanning to
find the next lines. This ensures that the right readings are obtained and recorded for calculating
the calibration factor. To calculate the relationship between the two points that have aligned, the
following formula is used:

Number of units = number of divisions on stage micrometer divided by the number of

divisions on the eyepiece.

When making measurements, it's particularly important that the right units of
measurements are used. Here, 1 mm is equal to 1000 micrometers (μm). The figures obtained
from the calculations should be converted to micrometers during measurements. For instance,
the diameter of the specimen here may be 14, 26 and 40 divisions. This number does not really
represent any specific units. In order to determine the length of the specimen, these units should
be multiplied to the conversion factor in order to get the measurements in micrometers. This
makes it possible to tell the actual length or width of the specimen being observed.

TASK 3

The extracellular matrix (ECM) is a non-cellular component found in all tissues and organs
that not only provides physical scaffolding for cellular constituents but also initiates critical
biochemical and biomechanical cues required for tissue morphogenesis, differentiation, and
homeostasis. The importance of the ECM is vividly demonstrated by the wide range of syndromes
caused by genetic abnormalities in ECM proteins, which can range from minor to severe. The
ECM, also known as connective tissue, is made up of three major classes of biomolecules. First,
there are structural proteins such as collagen and elastin. Second, there are specialized proteins
such as fibrillin, fibronectin, and laminin, and finally, there are proteoglycans, which are composed
of a protein core to which long chains of repeating disaccharide units known as
glycosaminoglycans (GAGs) are attached, forming extremely complex high molecular weight
components of the ECM.
 The dense bone tissue on the outside of the bone is known as compact bone. The
periosteum is a membrane that covers the entire bone and is where muscles and tendons attach.
The human body relies on compact bones for structural support and movement. Compact bone
is strong, forming protection and mechanical lever systems that facilitate movement. Compact
bone is composed of cylindrically shaped osteons that are linked together. Lamellae, lacunae,
and osteocytes are arranged around a central canal to form the osteon. Lamellae are the
extracellular matrix that surrounds the cells and provides hardness and rigidity to compact bone.
This layer matrix is composed of both organic and inorganic components. Tensile strength is
provided by collagen, while compressive strength is provided by hydroxyapatite crystals. lacunae
are gaps or empty spaces where there are osteocytes within it and lamellae. The central canal,
also known as the Haversian canal, is surrounded by concentric lamellae. Each central canal runs
parallel to the bone's long axis. It contains blood vessels, lymph vessels, and nerves. It also
contains a trace of connective tissue. Perforating canals, also known as Volkmann's canals,
intersect central canals at right angles. Perforating canals connect the blood vessels of the central
canal to the periosteum's blood supply. As observation, the extracellular matrix (ECM) is a
complex dynamic bio-environment with precise mechanical and biochemical properties. In
compact bone, ECMs regulate cell adhesion, proliferation, and responses to growth factors in
bone, as well as differentiation and, ultimately, the functional characteristics of mature bone. Bone
ECM can stimulate the formation of new bone by osteoblast-lineage cells such as MSCs,
osteoblasts, and osteocytes, as well as bone absorption by osteoclasts. The osteoinductive,
osteoconductive, and osteogenic potential of ECM-based scaffolds has received increasing
attention as bone regenerative medicine has advanced.

CONCLUSION

The hypothesis for this experiment is acceptable. It can be seen as compound

microscopes are very helpful to students in observing slides with its function and parts. For
example, a fine adjustment knob is used for small adjustments on focus. Students were able to
handle the microscope correctly and followed many precautions to protect the microscope for a
better observation. While using the oil immersion lens, students know that they should never use
it without the oil. This experiment shows that it is important to calibrate ocular micrometers with a
stage micrometer before making accurate measurements with a microscope. A stage micrometer
is simply a microscope slide with a scale etched on the surface. Their micrometer scale is 2 mm
long, with at least part of it etched with 0.01 mm divisions. By this experiment, students had
observed the extracellular matrix of compact bone. It is shown that ECMs regulate cell adhesion,
proliferation, and responses to growth factors in bone, as well as differentiation and ultimately the
functional properties of mature bone in compact bone.

References

1. Mokobi, F. (2022, September 17). Parts of a microscope with functions and labeled
diagram. Microbe Notes. https://microbenotes.com/parts-of-a-microscope/
2. Anderson. (2022, November 1). Microscope Stage Micrometer and Measurements.
MicroscopeMaster. Retrieved January 1, 2023, from
https://www.microscopemaster.com/microscope-stage-micrometer-and-
measurements.html
3. Micrometer Calibration | NY Microscope Co. (n.d.). Micrometer Calibration | NY
Microscope Co. Retrieved January 1, 2023, from
https://microscopeinternational.com/micrometer-calibration/
4. Lin, X., Patil, S., Gao, Y. G., & Qian, A. (2020, May 6). The Bone Extracellular Matrix in
Bone Formation and Regeneration. Frontiers. Retrieved January 4, 2023, from
https://www.frontiersin.org/articles/10.3389/fphar.2020.00757/full
 AS201
BACHELOR OF SCIENCE (HONS) BIOLOGY
CELL BIOLOGY BIO 411
LABORATORY REPORT 2
MADAM SARINA BINTI HASHIM

NAME NO MATRIC CLASS

MUHAMMAD IFFAT HANIF BIN MOHD RUDIN 2022899874 AS2011B

NUR FARRISYA BINTI MOHAMAD ZIN 2022478792 AS2011B

NURUL AIHAN BINTI AHMAD HILMI 2022697432 AS2011B

SHARIFAH NUR ALYA INSYIRAH BINTI SYED ADLAN 2022697594 AS2011B

OBJECTIVES

1. To observe and distinguish specialized cells in prokaryotic and eukaryotic organisms.

2. To study sperm morphology.
3. To become knowledgeable in performing staining techniques and its effects on cell
structure visualization.
4. To become familiarized with various stages of mitosis.
5. To become familiarized with the method of cell counting and determination of cell
concentration.

INTRODUCTION

New cells are formed in the body through cell division and then mature and become
specialized through cell differentiation. Cell differentiation results in a diverse range of cells that
become functionally specialized. The function they will perform is determined by their structure
and organization. This experiment will introduce students to the various types of plant cells. In
addition, students will be able to describe sperm morphology and understand the method for
determining sperm cell concentration.

In eukaryotic cells, the two types of cell division are meiosis and mitosis. These divisions
are for reproductive cells and somatic cells, respectively. Meiosis is a type of cell division in which
each daughter cell produced has half the number of chromosomes as the parent cell. Mitosis is a
type of division in which the parent cell divides into two new daughter cells that have the same
number of chromosomes as the parent cell.

MATERIAL & APPARATUS

1. Compound microscope
2. Slides and coverslips
3. Distilled water
4. Petri dishes
5. Immersion oil
6. 95% ethanol/methanol
7. Methylene blue
8. Giemsa stain
9. Coplin jars
 10. Pasteur pipettes
11. Clean cotton
12. Blood cell epithelium
13. Scalpels
14. Mouse sperm
15. Carnoy fixative
16. 18% hydrochloric acid
17. Onion bulb with growing roots
18. 2% toluidine blue/aceto-carmine/aceto-orcein
19. Cutting tiles
20. Razor blades
21. Forceps
22. Toothpicks

PROCEDURE

Task 1: Giemsa-stained blood smears

1. A finger was cleansed and swabbed with 70% alcohol. With a new sterile lancet, a
fingertip was punctured by pressing hard the lancet against the fingertip.
2. A drop of blood was placed on a clean glass slide. The blood was smeared and let it air
dry.
3. The dried smears have been fixed in absolute alcohol (ethanol or methanol) for 20
minutes.
4. The smear has been checked under the microscope.
5. The slide was stained for 20 minutes with Giemsa Stain. Then, the slide was rinsed with
distilled water at room temperature. Drain off the water and leave the slide to dry.
6. The slide has been observed under the microscope.
7. In order to stain Cheek cells, a small drop of distilled water has been placed in the
middle of a clean glass slide.
8. Cheek cells have been harvested by using a flat end of a toothpick to gently scrape the
inner side of cheeks.
9. The end of the toothpick was stirred in a drop of distilled water and throw away the
toothpick.
10. A smear has been made by using the same technique used to make a blood smear.
 11. Steps 3-6 were repeated with the cheek cell slide.
12. Observations has been recorded.

Task 2: Determining sperm concentration

1. A lab mouse (Mus musculus) has been euthanized prior to performing Task 2. The
reproductive organs of the mouse have been sketched and identified.
2. The testes of the mouse are excised and placed in a Petri dish. Lateral cuts were made
to the membrane of the epididymis in order to collect the sperms.
3. Sperms that have been collected were put onto a glass slide and covered with coverslip.
4. Microscopes were used to indicate the morphology of the sperm.
5. Observation has been recorded.

Task 3: Observation of mitosis in onion root apical meristems

1. 2cm from the cap of the root was measured and cut the root tip. The root was placed in
18% hydrochloric acid for 4 minutes.
2. It was transferred to the Carnoy fixative with chloroform (1:1) for 4 minutes.
3. The 2cm root tip was placed on a slide and trim away all but 1-2 mm of the tip was kept.
4. The tip was covered with 2-3 drops of aceto-orcein/acetocarmine/2% toluidine blue and
heated for 2 minutes.
5. The stain was bloated away, adding a drop of water, covered with a coverslip and
applying pressure to the coverslip with a pencil eraser until the cells in the tip spread out
as a single layer.
6. The prepared slide was prepared using a compound microscope. The low power
objective was used to identify thin layers of cells, and then using the high power
objective to observe mitotic stages in individual cells.
7. Observation has been recorded and labeled the stages of mitosis.
RESULT

TASK 1

Red blood cell

Red blood cells

400x total magnification

TASK 2

Sperm cell

Sperm cells
400x total magnification
TASK 3

Anaphase

Metaphase
Prophase

Onion root apical meristems

400x total magnification

DISCUSSION

In Task 1, blood cells are examined under a microscope, and the data indicates several
components in the blood. Blood is a liquid that flows through circulatory system arteries and
transports nutrients and oxygen to various cells. Blood cells mainly consist of red blood cells
(RBCs), white blood cells WBCs and platelets cells (Connie Rye, 2016). These cells are made of
bone marrow. They develop from stem cells into RBCs, WBCs, and platelets during their lives. In
addition, there are three types of WBC, which are lymphocytes, monocytes, and granulocytes,
and the main types of granulocytes are neutrophils, eosinophils, and basophils (Dean, 2005).

Erythrocytes, often known as RBCs, are specialized cells that travel throughout the body
and deliver oxygen to the tissues. The red blood cells are biconcave and small and do not have
mitochondria or a nucleus (Dean, 2005). These qualities enable RBCs to carry oxygen in their
intended function. Because there is no nucleus, the size and shape enhance the surface area-to-
volume ratio and provides more room for hemoglobin, which is essential for carrying oxygen. The
lack of mitochondria also prevents the RBCs from using the oxygen that they are transporting,
maximizing the oxygen delivered to the tissues (Connie Rye, 2016). WBCs, sometimes referred
to as leukocytes, vary from RBCs in that they play a larger role in the immune system. They lack
hemoglobin but have nuclei. Granulocytes and agranulocytes are the two basic types of white
blood cells. Neutrophils, eosinophils, and basophils are granulocytes, and when stained and seen
under a microscope, they have granules in their cytoplasm. Agranulocytes, which include
lymphocytes and monocytes, do not have granules in their cytoplasm. These kinds are crucial to
the defense. Some WBCs are involved in engulfing and breaking down pathogens, while others
recognise specific microorganisms and initiate immune responses against them (Connie Rye,
2016). Finally, the cell fragments known as platelets or thrombocytes are in charge of causing
blood to coagulate. Blood vessel lining damage causes platelets to be drawn to the site and form
a sticky clog.

One type of cell division known as mitosis occurs when a single cell splits into two
genetically identical progeny. Making sure that each daughter cell receives a complete set of
chromosomes is the goal of mitosis. As a result, during mitosis, the duplicated chromosomes are
meticulously divided up into neatly arranged phases. Interphase, prophase, metaphase,
anaphase, and telophase are the phases of mitosis. The phases are followed by cytokinesis,
which is the process of dividing the contents of the cell to make new cells (Connie Rye, Biology,
2016). The interphase is the initial stage. The cell is expanding in size and duplicates its DNA and
all of its internal components during this phase to prepare for division. The nuclear envelope is
still around the nucleus at this stage. The material around the nucleus is DNA in the form of
chromatin, and it does not stain well (Morrow, 2021). Then, during prophase, the nuclear envelope
begins to break down, the nucleolus disappears, and all of the chromosomes begin to gather
towards the cell's center, creating the conditions for chromosomal division. The mitotic spindle
begins to take shape. Their function is to organize the chromosomes and move them around
during mitosis (Connie Rye, Biology, 2016). As the cell prepares to split, the chromosomes align
at the metaphase plate and are pulled to either side by spindle fibers. The chromosomes are
subsequently separately pulled to the opposite side of the cell by shorter spindle fibers. This phase
is called anaphase. The nuclear envelope is then produced and the process of cytokinesis
overlaps in telophase, the last step. For plant cells, the cell wall is formed in the middle of the cell,
separating it into two daughter cells (Connie Rye, Biology, 2016).

In the experiment, two staining techniques are used to stain the samples. First off, Giemsa
stain is a staining method used to evaluate blood on both thin and thick smears. It was initially
developed to display malarial parasites in blood smears, but it is now utilized in histology for
normal blood smear testing. Methylene blue, azure, and eosin are the ingredients of the
differential stain. Giemsa stain has various applications in microbiology and pathology, such as
obtaining differential white blood cell counts, to differentiate nuclear and cytoplasmic morphology
of the blood cells like RBCs and WBCs (Rijal, 2019). Second, Aceto-carmine stain, a DNA-specific
stain, is employed to help in the visualization of various mitotic phases present in the cells of onion
root tips. A saturated carmine solution in 45% acetic acid makes up the stain. This staining
technique is usually applied for staining chromosomes of plant species, such as staining the
chromosomes of sugar beet (Tsuchiya, 1979).

CONCLUSION

In this experiment, the type of specialised cells that are being examined under a
microscope is determined. Since red blood cells lack a nucleus and are smaller than white blood
cells, but white blood cells have a nucleus and seem larger than red blood cells, it is easy to
distinguish between erythrocytes and leukocytes. Blood smears and the Giemsa staining
technique, which makes the cells more visible under the microscope, can be used to observe
these cells. We may become familiar with the laboratory process for calculating sperm
concentration using a hemocytometer by carrying out the experiment and counting the sperm
concentration using the correct formula. By treating the root tip-sample with acetocarmine to
increase contrast and colour the nucleic acid within, it may also be utilised to find actively dividing
tissues in plants. We can distinguish the mitotic phases of interphase, prophase, metaphase,
anaphase, and telophase in plants. At a 1000x magnification, all of the results can capture and
label every aspect of the cell's architecture.
REFERENCES

1. Connie Rye, R. W. (2016). Biology. Houston: OpenStax.

2. Connie Rye, R. W. (2016, October 21). Components of the Blood. Retrieved from
OpenStax: https://openstax.org/books/biology/pages/40-2-components-of-the-blood
3. Dean, L. (2005). Blood Groups and Red Cell Antigens. Retrieved from NCBI:
https://www.ncbi.nlm.nih.gov/books/NBK2263/
4. Morrow, M. (2021, May 28). Interphase, Mitosis, and Cytokinesis. Retrieved from
LibreTexts:
https://bio.libretexts.org/Bookshelves/Botany/Book%3A_A_Photographic_Atlas_for_Bota
ny_(Morrow)/10%3A_Cells_and_Tissues/10.03%3A_Cell_Division/10.3.01%3A_Interph
ase_Mitosis_and_Cytokinesis
5. Rijal, N. (2019, July 13). Giemsa Stain: Principle, Procedure, Results. Retrieved from
Microbe Online: https://microbeonline.com/giemsa-stain-principle-procedure-and-results/
6. Tsuchiya, T. N. (1979). Acetocarmine squash method for observing sugar beet
chromosomes. Euphytica, 249-256
 AS201
BACHELOR OF SCIENCE (HONS) BIOLOGY
CELL BIOLOGY BIO 411
LABORATORY REPORT 3
MADAM SARINA BINTI HASHIM

NAME NO MATRIC CLASS

MUHAMMAD IFFAT HANIF BIN MOHD RUDIN 2022899874 AS2011B

NUR FARRISYA BINTI MOHAMAD ZIN 2022478792 AS2011B

NURUL AIHAN BINTI AHMAD HILMI 2022697432 AS2011B

SHARIFAH NUR ALYA INSYIRAH BINTI SYED 2022697594 AS2011B

ADLAN
OBJECTIVES

1. To study the process of osmosis in plant cell

2. To determine the effect of surface area to volume ratio on osmosis
3. To determine the tonicity of the surface solutions as hypotonic, isotonic and
hypertonic based on experimental observations.

INTRODUCTION

Molecules interact and move in new directions due to kinetic energy, a source of energy
stored in cells. This contact results in diffusion. The random movement of molecules from a higher
concentration area to a lower concentration area is known as diffusion. Diffusion is a sort of
osmosis. This occurs when water diffuses through a membrane that is selectively permeable from
one area with a greater water potential to another with a lower water potential. The amount of
water's free energy in a solution is measured as water potential. Water potential predicts the
movement of water into or out of plant cells. Since sucrose is a hypertonic solution, meaning that
it has a greater number of particles dissolved in the solution than a hypotonic solution like water
where there are less particles dissolved, the difference in mass of potato will increase as
concentration of sucrose increases. When the potato is placed in a concentration of sucrose
where the water potential is greater than that of the potato, particles will move from the potato to
the sucrose and there will be a loss of mass, whereas if it is placed in a concentration where they
have the same water potential, the solution is known as isotonic.

MATERIALS
1. Potato
2. Onions scales
3. Glass slide
4. Scalpels
5. Digital balance
6. String ruler
7. Sucrose solutions of varying molarity: 0.01M, 0.02M, 0.05M, 0.10M, 0.30M and
0.50M
8. Tissue paper
 9. Test tubes
10. Petri dish
11. Forceps
12. Laboratory cork borer

PROCEDURE

TASK 1

1. Potato cylinder has been cut out by using the laboratory cork borer in order to produce
uniform shape
2. Potato cylinder was divided into 15 cutting for six plates
3. The initial mass of each plate was weighed on digital balance.
4. Seven petri dishes were labeled and filled with sucrose solution which are 0.05M, 0.02M,
0.01M, 0.1M, 0.3M, 0.5M and distilled water.
5. All the 15 cuttings were immersed into the sucrose solution.
6. After an hour, all the potato from each plate was dried with tissue paper
7. Mass after potatoes was weighed
8. The result was recorded.

TASK 2

1. 7 slides for observation were prepared by removing the onion epidermis from the onion
scale. The onion epidermis chosen was the transparent membrane lining the inner side
of the onion scale.
2. The epidermis was removed by using forceps and the onion epidermis was laid flat on a
glass slide.
3. The slides were labeled with 0.05M, 0.02M, 0.01M,0.3M, 0.5M and distilled water to
represent the molarity of the sucrose solution.
4. A drop of the sucrose solution with the respective molarity was placed on the slide of the
onion epidermis. The slide was observed immediately.
5. The steps for the consecutive molarity of sucrose and distilled water were repeated.
6. Observation by photographing was recorded.
RESULT

TASK 1
Control parameter: Time taken(hour)
Measured parameter(s): The change in weight of potato cylinders (grams)
Variables measured: Concentration solution of sucrose (M)

Concentration solution of 0.01 0.02 0.05 0.10 0.30 0.50

sucrose (M)

Weight of the Initial 3.6 3.3 3.3 3.5 3.4 3.1

cell potatoes
Final 3.9 3.4 3.3 3.0 2.9 2.6
(gram)

Difference 0.3 0.1 0.0 -0.5 -0.5 -0.5

TASK 2

Nucleus

Onion Epidermis
100x total magnification
Treatment: 0.01M sucrose solution
 Nucleus

Figure 2: Onion Epidermis

100x total magnification
Treatment: 0.50M sucrose solution

Cell wall Nucleus

Figure 3: Onion Epidermis stain

100x total magnification
Treatment: 0.01M sucrose solution
DISCUSSION

Movement of a solvent, such as water, into a solution with a higher solute concentration
across a semipermeable membrane, such as that found in a live cell, tends to equal the
concentrations of solute on the two sides of the membrane. In this experiment, the potatoes were
soaked in a petri dish full of sucrose solution. As we know, sucrose solution is an extremely weak
base, but sucrose solution can be hypertonic, hypotonic or isotonic it depends on the
concentration of the sucrose solution.

The tonicity process includes, which is somewhat similar to the osmotic pressure process
but flows in the opposite direction. To put it more precisely, the concentration of dissolved solutes
in solution and the cell's membrane permeability are the two main variables that affect the
direction of the diffusion process. Tonicity is the ability of the solution to cause the water to flow
in or out of the cell through osmosis. The terms hypertonic, hypotonic, and isotonic are all used
to describe tonicity. Because the solute concentration is much higher outside the cell than inside
it, in a hypertonic solution, the water flows out of the cell and the cell contracts. In a hypotonic
solution, the water enters the cell and causes it to swell since the concentration of the solutes is
considerably lower than within the cell. Last but not least, there will be no movement of water into
the cell because both solute concentration and the cell’s volume are already stable, called isotonic
solution. From the experiment, when the potatoes were placed into concentration solution of
sucrose 0.01M and 0.02M, the potatoes become turgid because of the solution is a hypotonic
solution, which means the water concentration of the solution is way greater than inside the
potatoes, as the sucrose concentration inside the potatoes is so high. So, the water flows into the
potatoes and makes the potatoes increase in size. When the potatoes were placed into the
concentration solution of sucrose 0.10M,0.30M and 0.50M, the potatoes shrink because the
sucrose concentration is the hypertonic solution. For 0.05M concentration solution of sucrose, the
volume of the cell will remain constant and there won't be any net water inflow or outflow. The
solution is isotonic to the cell if the concentration of solutes within and outside the cell are equal,
and the solutes cannot pass the membrane.
CONCLUSION

This last outcome is a result of osmosis. Osmosis is a special kind of diffusion; it refers to
the diffusion of water. Osmosis behaves differently depending on the solution a cell is in. Water
will always move in the direction that will achieve dynamic equilibrium –an equal balance of
solution on both sides of the membrane. For example, in a hypertonic solution, or one that has a
high concentration of solute, water will move out of the cell, causing it to shrivel up. The opposite
occurs in a hypotonic solution; water will rush from a diluted solution across a membrane, causing
cells to swell and sometimes burst. If it does not burst, it will eventually reach dynamic equilibrium.
As the results show, the potato chores did grow in low sucrose concentration and did shrink in
high sucrose concentration. This also gives another fact that if the potatoes were to be measured
in weight, we would then see that the mass would decrease in a high sucrose concentration and
decrease in a low sucrose concentration/distilled water.

REFERENCES

1. Britannica, T. Editors of Encyclopedia (2022, September 30). osmosis. Encyclopedia

Britannica. https://www.britannica.com/science/osmosis
2. Merriam-Webster. (n.d.). Osmosis. In Merriam-Webster.com dictionary. Retrieved
December 30, 2022, from https://www.merriam-webster.com/dictionary/osmosis
3. Tonicity: hypertonic, isotonic & hypotonic solutions (article) | Khan Academy. (n.d.).
Khan Academy. Retrieved December 30, 2022, from
https://www.khanacademy.org/science/ap-biology/cell-structure-and-
function/mechanisms-of-transport-tonicity-and-osmoregulation/a/osmosis
4. Asghari. (2017, October 1). The effect of osmosis on potatoes in different
concentrations of sucrose solutions. Athenology. Retrieved January 1, 2023, from
https://www.athenology.com/post/2017/09/23/the-effect-of-osmosis-on-potatoes-in-
different-concentrations-of-sucrose-solutions