Name: Date:

Program and Section:

Laboratory Activity No. 3

ENTRY AND EXIT OF SUBSTANCES IN A CELL

Introduction

Outside the cell membrane lies the environment of the cell. Thus, the membrane is the
boundary that separates the units of living material from the external environment and its
neighbour cells. Through the cell membrane, then, must pass everything that the cell gets from
the environment and through it also must pass everything that the cell returns to the
environment as long as the cell is alive.

Objective
In this activity you will:
1. Perform diffusion, dialysis and osmosis; and
2. Discuss the biochemical importance of diffusion, dialysis and osmosis.

Material/ Reagents
KMnO4 petri dish
0.9% NaCl solution beaker
Chicken/ Beef bouillon graphing paper
Distilled water test tubes
Gastric lining microscope
Defibrinated blood glass slides and cover
1% AgNO3 slips string
6 m NaOH timer
1% CuSO4 thermometer
0.2% ninhydrin solution ruler
2% NaCl solution

Procedure
A. Diffusion
1. Put about 25 mL of distilled water is a petri dish over a piece of graphing paper. Get
the temperature of the water and let it stand undisturbed.

Temperature of water:

Place 2-3 KMnO4 crystals at the bottom of the petri dish and record the time required for
the colored ions to travel 1 cm along line toward the center.

Time of travel of ions at room temperature:

3. Discard the contents of the petri dish.

4. Get the beaker and put 25 mL of water. Heat the water to a temperature of 80-90 C and
place it over a graphing paper.

5. Again, add 2-3 KMnO4 crystals at the bottom of the beaker and record the time
required for the colored ions to travel 1 cm along line toward the center.
 Time of travel of ions at elevated Temperature:

6. Account for your observations.

7. What is the biochemical importance of this process? Give the example.

B. Dialysis
1. Get a 10 cm-long gastric lining of about 2.5 cm in diameter.

2. Close on end of the gastric lining using 1 string.

3. Pour about 20 mL of chicken bouillon on the gastric lining and close the other end.
Now wash the outside portion of the gastric lining with distilled water.

4. Place the gastric linin gin a beaker and add distilled water to immerse about three
fourths of it. (Note: check the distilled water for chloride content by adding 1 mL of 1%
AgNO3. The result must be colorless.)

5. Let it stand for 1 hour at room temperature.

6. Prepare the following test on the dialysate.

a. Test for chloride ion: to 1 mL of dialyzate, add ` mL of AgNO3.
What did you observe? What does your observation suggests?

b. Test for Protein: to 1 mL of dialyzate, add 1 mL of 6 M NaOH and 5 drops of

1% CuSO4.
What did you observe? What does your observation suggest?

c. Test for amino acid: to 1 mL of dialyzate, add 1 mL of 0.2% ninhydrin solution.

Heat in a water bath until a blue-violet color appears.
How will you explain your result?
C. Osmosis
1. Place 5 drops of Defibrinated blood in each of the four test tubes.

2. add 20 drops of distilled water to the first test tube, 20 drops of 0.9% NaCl to the
second and 20 drops of 2% NaCl to the third leaving the fourth as the control

3. Let it stand for 2 minutes.

4. Shake and examine the appearance of the blood cells from each test tube under the
microscope. Draw and compare.

Supplementary Questions
1. Differentiate active transport from the passive transport.

2. Draw and describe the movement of Na+ and K+ ions in the cell.

3. Give an operational definition of the following:

a. Diffusion
b. Dialysis
c. Osmosis