CELL RESPIRATION LAB

BACKGROUND MATERIAL

Cell respiration is the oxidation of glucose by the addition of oxygen,

accompanied by the production of ATP. Cells which are metabolically active must
consume O2 in order to produce ATP, which is used by cellular processes. The
equation for cell respiration is:

C6 H12O6 + 6O2 ---> 6H2O + 6CO2 + energy (ATP)

In addition to the essential ATP, carbon dioxide is produced as a waste gas.

THE EXERCISE

This exercise utilizes a respirometer to measure the amount of oxygen being

utilized by cells for respiration. The respirometer allows for an indirect measurement of
the rate at which cells perform cell respiration. Respiration rate of germinating peas will
be compared to the rate for dormant (nongerminating) peas in order to learn more about
the need of oxygen under different metabolic conditions.

(Respirometer set up; in our example pea seeds will be used)

 Potassium hydroxide (KOH) will be used in this experiment since it reacts with
CO2 to remove the gas from the respirometer by converting it to a precipitate -
potassium carbonate (K2CO3 ). Once the gas is transformed into a solid, a vacuum is
created in the respirometer which will be filled by water entering the apparatus. The
degree to which water enters the respirometer is directly proportional to the amount of
carbon dioxide being produced; therefore, oxygen consumption by the cells can be
evaluated by the movement of water into the respirometer.

Oxygen consumption will be measured over time by three respirometer

assembled and maintained as follows:

Respirometer Temperature Contents

1 Room Germinating Seeds

2 Room Dry Seeds and Beads

3 Room Beads

PROCEDURE

1. Set up a room temperature water bath by filling the black plastic pan with water.

2. Respirometer 1: Fill a 100 ml graduated cylinder to the 50 ml mark, and add 25

germinating peas. Note how much water was displaced by the peas : _____________.
Pour out the peas and place on a paper towel for later use.

3. Respirometer 2: Fill a 100 ml graduated cylinder to the 50 ml mark, and add 25 dried
peas. Add glass beads to the cylinder until the amount of water displaced is equal to
that of respirometer 1. Pour out the peas, dry, and place on a paper towel for later use.

4. Respirometer 3: Fill a 100 ml graduated cylinder to the 50 ml mark, and add glass
beads until the volume of water displaced is equal to that displaced in respirometer 1.
5. Assemble the three respirometers as follows: Obtain three vials and three stoppers.

A. In respirometer 1, place 10 pellets of KOH (NOTE - KOH is a strong base so use

forceps to transfer the pellets to the vial). Next put a small wad of dry cotton into the
vial. Finally put the germinating peas into the vial and stopper. Refer to figure 7.2 (p.
80) in lab book to see the respirometer assembly.
B. In respirometer 2, add 10 pellets of KOH, cotton (same amount as in respirometer
1) and dry peas/glass beads. Stopper the vial.
C. In respirometer 3, add 10 pellets of KOH, cotton and glass beads. Stopper the
vial.

6. Make a sling across the black plastic pan to support the calibrated pipette attached to
the stopper. Lay the vials in the water, ensuring the pipette opening is not in the water
but resting on the tape sling. Leave the vials in this position for 7 minutes to allow for
equilibration. While waiting, obtain another pipette and familiarize yourself with the
graduation markings so that you can effectively read the pipette once the experiment
begins.

7. After equilibration, cut the tape so that the pipettes are parallel to the bottom of the
pan and submerged. Quickly twist the pipette so that it is in a readable position
readable, and then remove your hands from the water. Water will begin to move into
the tube, but will stop for a time. If water quickly moves into the pipette, and doesn't
stop take the vial out and check for leaks.

8. Allow 3 more minutes to pass for additional equilibration, then record, to the nearest
0.01 ml, the initial position of water in each pipette (time 0). Every 5 minutes for 20
minutes take readings of the water's position in each pipette and record.

LAB GROUP ASSIGNMENTS

Each lab group will prepare the three respirometers described above and collect the
data.
Time Reading

Initial - 0 .

5 min .

10 min .

15 min .

20 min .

Respirometer 1 - Germinating Peas

Time Reading

Initial - 0 .

5 min .

10 min .

15 min .

20 min .

Respirometer 2 - Dry Peas and Beads

Time Reading

Initial - 0 .

5 min .

10 min .

15 min .

20 min .

Respirometer 3 - Beads Alon

ANALYSIS OF RESULTS

1. Graph the data projected. Average Reading for each group is on the Y (vertical)
axis, and the time data should be placed along the X (horizontal) axis.

2. State a hypothesis for this experiment.

3. Identify the control for the experiment.

4. What conclusion can be drawn from the data collected?