HALIC UNIVERSITY

FACULTY OF ARTS AND SCIENCES

DEPARTMENT OF MOLECULAR BIOLOGY AND GENETICS

MBG104
GENERAL BIOLOGY LABORATORY-II

INSTRUCTOR
Doc. Dr. Sahra KIRMUSAOĞLU

EXPERIMENT II
THE PROPERTIES AND INVESTIGATION OF THE CELL
MEMBRANE (PART II)

ASSISTANTS
Res. Assist. Anıl CEBECİ
Res. Assist. Hatice KURNAZ
Res. Assist. Şafak ŞENER

Experiment Date: 25.03.2022

Name of the Student

Hatice Sena YENİGÜN

GROUP MEMBERS
Helin Dila TARHAN
Nesibe DİLMAÇ
Bilge Naz YILMAZ

Submission Date of The Report: 01.04.2022

AIM
All living things, from cells to large organisms, want to maintain their homeostasis in order to
survive. In this experiment, the changes created by the external environment concentration
differences (tonicity) in plant and animal cells will be observed.

INTRODUCTION
We have to take into account the concentration of the solution in a cell or a membrane-
separated environment, that is, the surrounding solution, and the permeability of the
membrane. Briefly, tonicity, which is a ratio of the concentration percent of two solutions
separated by a semipermeable membrane, is the ability of the solution around the cell to cause
it to gain or lose water. In other words, the ratio of the concentrations of solute molecules in
membrane-separated solutions to each other is called tonicity.
If the solute concentration is too high to pass through the membrane in the solution outside
the cell, the water inside the cell will tend to leave the cell, or vice versa.
Conditions and environments where the concentration ratio is equal in the intramembrane and
extramembrane media are called isotonic (iso means "same"). For example, if a cell, such as
an animal cell, is in an isotonic environment, there is no clear water passage through the
plasma membrane, only water diffuses through the membrane, and this transition continues at
equal speed in both directions. The volume of the animal cell is stable as long as it is in this
environment.
The environment with a higher concentration of the extracellular environment than inside the
cell is called hypertonic (hyper is "more"; here it is expressed that the amount of solute
that cannot pass through the membrane is high). For example, if we put an animal cell in a
hypertonic environment, we will witness that the cell loses water and shrinks, and if not
intervened immediately, it may even result in death. As an example, let's consider the
creatures that live in any lake. If the salinity of the water in the lake becomes hypertonic
compared to the cells of living things, the cells may shrink and eventually death may occur. In
another example, if we assume that we put the plant cell in a hypertonic environment, the
plasma membrane shrinks to separate from the cell wall, and this is called plasmolysis. It is a
condition that causes the plants to wilt.
In another medium, the concentration outside the membrane is less dense than the
concentration inside the membrane, and this environment is called hypotonic (hypo means
"less"). If we put an animal cell in a hypotonic environment, water molecules will enter the
cell faster than the concentration in the environment. This will cause the animal cell to first
inflate and then burst, like a water balloon, and this event is called lysis (the disintegration of

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blood cells by swelling is called hemolysis). In a hypotonic environment, a plant cell does not
disintegrate due to its cell wall, but the central vacuole grows and swells and the cell wall
exerts pressure on the cell wall due to this swelling, this pressure is called turgor pressure.
Turgor pressure is a factor that causes plants to stand upright.

MATERIALS

Chemicals
0.9% NaCl solution
10% NaCl solution
0.35% NaCl solution
40% sucrose solution
Distilled Water
Biological Material
Human blood sample
Elodea

Laboratory equipment
Pliers
Glass pen
Pasteur pipette
Slide
Coverslip
Micropipette
Capped test tube
Written paper

Devices
Light microscope

METHODS
A. PLASMOLYSIS IN ELODEA CELLS
 2 slides are taken and numbered.
 A sample of elodea is placed on each of the 2 slides.
 1 drop of distilled water is dropped on slide 1 and covered with a coverslip and
examined under a light microscope under x4, x10 and x40 lenses.
 1 drop of 40% sucrose solution is dripped onto slide 2 and covered with a
coverslip and examined under a light microscope under x4, x10 and x40 lenses.

B. TONICITY IN RED BLOOD CELLS

 3 capped test tubes are numbered 1,2,3 with caps.
 5mL of 0.9% NaCl solution is put into the first tube with the help of a micro
pipette.

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  5mL of 10% NaCl solution is put into the second tube with the help of a micro
pipette.
 5mL of 0.35% NaCl solution is put into the third tube with the help of a micro
pipette.
 Add 5 drops of human blood sample to each test tube using a micro pipette.
 Cover the test tubes and mix gently and slowly.
 A slide is taken and divided into 3 equal sections with a permanent marker and
numbered 1,2,3.
 A drop from the first tube on the first area on the slide, a drop from the second
tube on the number 2 area, a drop from the third tube on the number 3 area is
covered with a coverslip.
 The prepared slide is examined under a light microscope with x10 and x40
lenses.

RESULTS
EXPERIMENT I
PLASMOLYSIS IN ELODEA CELLS

Elodea with distilled water Elodea with 40% sucrose solution

X40 x40

QUESTIONS
1. Compare the central vacuoles and plasma membranes of Elodea leaf cells.
2. What are the differences you observed?
3. Was the 40% sucrose solution hypertonic, isotonic or hypotonic compared to the
cytoplasm?
ANSWERS
1) The structure that acts as a barrier between the cell and organelles and their
environment is called the plasma membrane. It consists of lipids, proteins and
carbohydrates. They provide cell-cell communication. Fluid-filled sacs surrounded by
a single-layered membrane are called vacuoles. A central vacuole is a type of vacuole
found in plant cells but not in animal cells. The central vacuole serves to hold water in

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 the plant cells, and the water it holds passes through the plant, increasing the turgor
pressure within the plant. That is, water acts on the cell wall as pressure.
The tonoplast surrounds the central vacuole. The only difference between tonoplast
and plasma membrane is that tonoplast surrounds the central vacuole of the plant cell
while plasma membrane is the membrane that surrounds the cytoplasm of a cell.
2) In the isotonic medium, the cell wall and cell wall were together, and the central
vacuole was of average size. In the hypertonic environment, the central vacuole
became too small to be observed and the plasma membrane moved away from the cell
wall. In the hypotonic environment, the central vacuole enlarged to almost the entire
volume in the center of the cell, and the plasma membrane swelled to exert pressure
on the cell wall.
3) 40% sucrose solution is hypertonic compared to cytoplasm.
EXPERIMENT II
TONICITY IN RED BLOOD CELLS

0.9% NaCl and blood 10% NaCl and blood 0.35% NaCl and blood
x40 x40 x40

Table 1. The effects of solute concentration on red blood cells

Number of Tube Content Legibility of the Tonicity of Solution
Text
1 5mL of 0.9% NaCl - Isotonic
and 5 drops of blood
2 5mL of 10% NaCl and - Hypertonic
5 drops of blood
3 5mL of 0.35% NaCl + Hypotonic
and 5 drops of blood

QUESTIONS
1. Which test tubes have healthy cells? Explain the reason.
2. Explain what happens when Elodea cells and red blood cells are placed in a hypotonic
environment.
3. Summarize the tonicity event using examples of Elodea and red blood cells.

ANSWERS

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 1) Cells in tube 1 are healthy, providing an isotonic environment for the blood cell. Because
the rate of entry and exit of water molecules in the plasma membrane is equal.
2) In a hypotonic environment, red blood cells swell and break down (hemolysis).
However, elodea cells with a cell wall do not lyse, the cell swells and the plasma
membrane exerts pressure on the cell wall (turgor pressure).
3) The ratio of the concentrations of solute molecules in solutions separated by a membrane
is called tonicity. For example, in the blood cell experiment, in the tube containing 10%
NaCl solution and 5 drops of blood, the solution had a lower solute concentration from
inside the cell. For this reason, the solution was called hypotonic with respect to the cell.

DISCUSSION
There is nothing to argue about this experiment. In environments with different tonicity ratios,
plant and animal cells were examined and it was observed that in hypertonic environment,
elodea cells, in which blood cells shrank, undergo plasmolysis, and in hypotonic environment,
elodea cells, in which blood cells undergo hemolysis, exert turgor pressure on the cell wall of
the plasma membrane, where the central vacuole grows.

REFERENCES
1) Kırmusaoğlu,S.Genel Biyoloji Laboratuvar Klavuzu.Ankara,Hipokrat Yayınları,2018.
2) Audesirk, T., Audesirk, G., & Byers, B. E. (2014). Biology: Life on earth with physiology.
Harlow: Pearson Education Limited.