PRACTICAL 3: TRANSPORT ACROSS MEMBRANE Objectives: The objectives of this experiment are: 1.

To determine the osmotic pressure of a cell 2. To determine the osmotic pressure in the atmosphere from the graph 3. To determine the concentration of sodium chloride solution that causes haemolysis 4. To determine the concentration of sodium chloride that is isotonic to red blood cells. Introduction: The cell membrane is a selectively permeable structure because only selected materials can pass through it. Water molecules can easily pass through the membrane and the movement of water is called osmosis. The direction of movement of water molecules is determined by the concentration of the solutes on both sides of the membrane. The water potential inside and outside of the cell is said to be isotonic, that is the movement of water molecules in both direction is at the same rate. The membrane vacuole is also a semi-permeable structure and the condition in the vacuole is isotonic to the cell environment. In hypertonic environment, water molecules will move out of the cell and the cell shrinks. The shrinking of cell due to the hypertonic environment happens in plant and animal cells. The shrinking plant cell is called plasmolysis while the shrinking of animal cell is called crenation. When a plant cell is in a hypotonic environment, it will expand but the increase in size is restricted by the cell wall (turgid). On the other hand, animal cells which are in the hypotonic environment will expand and burst and this is called lysis or haemolysis.

Materials and method: EXERCISE 3.1: OSMOSIS PRESSURE Apparatus Boiling tube Beaker Petri dish Measuring cylinder (25ml) Forceps Pipette Cork border Fresh potato tuber Sucrose solutions 1.0M (40ml per group) Distilled water Labeling paper Graph paper Razor blade Tile Ruler Filter paper Materials

Procedure and observation: 1. 20ml of sucrose solution with different molarities is prepared using the dilution method. The molarities required are 0.2M, 0.3M, 0.4M and 0.5M. The volume of the sucrose solution (1.0M) and distilled water used are recorded in the table below. Every tube is labeled. Molarity Volume of 1.0M sucrose (ml) Volume of distilled water (ml) 18 16 14 12 10 2 4 6 8 10 0.1M 0.2M 0.3M 0.4M 0.5M

2. 15 pieces of potato strips are prepared using cork border. The length of each strip is approximately 4cm-6cm. The average length of each strips are recorded. 3. The lengths of three strips of potato are recorded and are put into the tube. This step is repeated for the other concentration of sucrose solutions. 4. After 30 minutes, the three strips are removed from each boiling tube, wiped and immediately the final length of each strip is recorded in the same table. 5. The average lengths of potato strips are determined.

6. Using a graph paper: i. To determine the osmotic pressure, a standard graph of osmotic pressure against the molarity of sucrose solution is drawn.

Molarity (M) Osmotic pressure (atm)

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

0.55

1.3

2.6

4.0

5.3

6.7

8.1

9.6

11.1

12.6

14.3

16.0

ii.

A graph is drawn to show the changes in length of the potato strips against the molarities of the sucrose solutions.

7. From the graphs: i. ii. The osmotic concentration of the potato tissue in Molar sucrose at a hypertonic, isotonic and hypotonic are determined. The pressure in atmosphere is determined.

EXERCISE 3.2: HAEMOLYSIS Apparatus 6 test tubes Pipette 10ml Glass rod Gloves Compound microscope Slides Cover slips Sterilized lancet Cotton Dropper Materials 40ml of 1% NaCl solution Distilled water Ethanol Oil immersion

Procedure and observation: 1. 6 tubes are labeled from A to F. 2. 6 solution of NaCl with different concentration from the stock solution of 1% NaCl are prepared as shown in the table below. Concentration of NaCl (%) Volume of 1% NaCl (ml) Volume of distilled water (ml) Total volume (ml) 1 10 0 10 0.8 8 2 10 0.6 6 4 10 0.4 4 6 10 0.2 2 8 10 0 0 10 10

3. The hand is cleaned using ethanol. One of the fingers is pricked using a sterilized lancet. Then, the lancet is disposed. 4. 2 drops of blood are added into each test tube. 5. Each tube is shaken and leaved for 5 minutes. 6. The colour of the solution from each tube is examined. 7. A drop of the solution from each tube is transferred onto a slide. 8. A cover slip is placed onto the slide and examined under 100x magnification. 9. All the observation is recorded in the table.

Results: EXERCISE 3.1: Osmotic pressure Molarity Volume of 1.0M sucrose (ml) Volume of distilled water (ml) EXERCISE 3.2: Haemolysis Test Tubes Concentration of NaCl (%) Colour of solution (clear/cloudy) A B C D E F 1.0 0.8 0.6 0.4 0.2 0.0 Clear Clear Clear Cloudy Cloudy Cloudy Appearance of erythrocyte 18 16 14 12 10 2 4 6 8 10 0.1M 0.2M 0.3M 0.4M 0.5M