Department of Plant Sciences/ University of Colombo

BT1009 CELL BIOLOGY PRACTICALS - 2023

BT1009 CELL BIOLOGY PRACTICALS

(Laboratory Sessions)

Practical 01
Permeability of Cell Membranes and Movement of Water molecules Across Cell Membranes

Objectives:
1. To study the effect of different external solute concentrations on osmosis in animal cells
(RBCs) and plant cells (Rhoeo sp.).
2. To study the effects of various chemicals on permeability control of plant cells.

Experiment 1A
The effect of different external solute concentrations on red blood cells

Red blood cells are rather opaque discs which transmit very little light when suspended on a liquid.
When the cells haemolyse, the suspension will become transparent. This characteristic allows one to
determine the tonicity of various solutions to red blood cells and direction and rate of osmosis. If a water
suspension is made of red blood cells and the test tube containing the suspension is placed against this
page, the printing will become legible after a certain time. At that time (when the print is legible)
approximately 75% of the cells will have haemolysed.

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 Department of Plant Sciences/ University of Colombo
BT1009 CELL BIOLOGY PRACTICALS - 2023

Steps

[Important: Wear gloves when handling blood]

1. Use test tubes (labelled experiment 1A) containing 5 ml of each of the following solutions,
1) 0.043 M NaCI 2) 0.155 M NaCI 3) 0.860 M NaCI

2. Working with only one test tube at a time, add two drops of blood, mix well by inverting the
tube several times and record the time to the nearest second.

3. Place the tube against a printed page and record the time it takes for the printing to become
legible (clear enough to read). Use the second hand of your watch, the timer, or the wall clock
to record the haemolysis time in each tube. If haemolysis does not occur within one minute,
proceed to the next tube. Be sure, however, that you record the time at which you added blood
to the previous tube.

Results & Observations

Haemolysis time for red blood cells in different NaCl concentrations.

Solution Haemolysis time (s) Net movement of water

1) 0.043 M NaCl
2) 0.155 M NaCl
3) 0.860 M NaCl

a. Which concentration of NaCl is hypotonic to the red blood cell? ……………………………………….

b. When a drop of blood diluted with 0.155 M NaCl (A) and 0.860 M NaCl (B) was observed
under the light microscope they appeared as follows,

A - 0.155 M NaCl B - 0.860 M NaCl

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 Department of Plant Sciences/ University of Colombo
BT1009 CELL BIOLOGY PRACTICALS - 2023

Which concentration of NaCl is:

a. Most nearly isotonic to the red blood cell?................................................

b. Hypertonic to the red blood cell? ...............................................................

Experiment 1B
The effect of varying external solute concentrations in plant cells

Plant vacuoles may take up 90% of the volume of a cell and contain water and dissolved
substances that usually make a plant cell hypertonic in relation to the surrounding medium
(which is then hypotonic). The vacuole takes in water by osmosis, swelling as it does so and
exerting pressure on the cytoplasm which in turn presses against the cell membrane and cell wall.
In mature cells, the cell wall is under constant water pressure known as turgor. Turgor pressure
plays a major role in keeping leaves and stems firm or turgid. With water loss, turgor pressure is
reduced in the cells of a plant and the plant wilts.
If a plant cell is placed in a hypertonic solution, the vacuole loses water by osmosis and the
vacuolar membrane and plasma membrane shrink away from the cell wall. This condition is
known as plasmolysis.

Steps

1. Make a wet mount of Rhoeo (Add a drop of 0.860 M NaCl solution instead of water). Make
observations of cell shape. Can you distinguish the plasma membrane, cell wall and area of
the central vacuole?

2. Exchange the salt solution for distilled water by lifting the cover slip and flooding the slide
with distilled water. Observe closely for changes that occur in the cell and make a sketch.

Results & Observations

1) Sketch and make notes on the appearance of Rhoeo cells mounted in,
i. Distilled water
ii. 0.860 M NaCl

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 Department of Plant Sciences/ University of Colombo
BT1009 CELL BIOLOGY PRACTICALS - 2023

i. ……………………………………………………………
……………………………………………………………
……………………………………………………………
……………………………………………………………
……………………………………………………………
……………………………………………………………
……………………………………………………………

Tonicity of external solution –

……………………………………………………………

ii. ……………………………………………………………
……………………………………………………………
……………………………………………………………
……………………………………………………………
……………………………………………………………
……………………………………………………………
……………………………………………………………

Tonicity of external solution –

……………………………………………………………

Experiment 1C
Loss of permeability control
The beet-root strips provided have been well washed to remove the red pigment betacyanin
from damaged cells.
Steps
Place a fresh strip of beet root tissue in 5 ml of each of the following:

a) 2% sugar solution b) 95 % Ethyl alcohol c) 0.1 N HCl d) SDS (10%)

Observe after 30 min. Test whether the strip is turgid or flaccid by squeezing between the
fingers. In each case record your observations.
Wash the strips well and place them in about 10 ml of water. Once again test the turgidity /
flaccidity after 30 min. Record your observations.

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 Department of Plant Sciences/ University of Colombo
BT1009 CELL BIOLOGY PRACTICALS - 2023

Results & Observations

Solution Turgidity/ Flaccidity after Other observations Turgidity/ Flaccidity

30 min. in solution after 30 min. in water

2% sugar solution

0.1 N HCl

SDS

95% C2H5OH

Which solutions caused irreversible loss of permeability control? Explain your observations and
conclusion/s.
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