Biology 101

Lab
Cellular Respiration
The online student should read this lab before answering the questions, completing the table, and completing the graphs.

Cells store energy in the chemical bonds of organic molecules such as fats and sugars. This energy can be released through a
process of oxidation, much like the energy stored in gasoline is released as the fuel is burned in the internal combustion engine of
an automobile.
Just as the energy released from the combustion of gasoline is used to power an automobile (do mechanical work), the energy
released by oxidation of organic molecules in the cell is used to do cellular work. A cell cannot use this energy directly, however.
The released energy must first be harnessed in the form of the molecule ATP, which acts as the “energy currency” of the cell.
The production of ATP by oxidation of organic molecules is a multi-step process, with each step under the control of a specific
enzyme.

The most energy-efficient form of the oxidation of organic molecules is termed cellular respiration. Cellular respiration is an
aerobic process (that is, one that requires oxygen). Most cells rely mainly on the oxidation of glucose (a sugar) as the fuel for
production of ATP by cellular respiration. The process by which glucose is oxidized to carbon dioxide (CO 2) and water (H2O) to
generate ATP can be represented by the following chemical equation:

36 ADP + 36 Pi + 6O2 + C6H12O6 ⎯⎯→ 6CO2 + 6 H2O + 36 ATP

This process yields a net of 36 ATP molecules for each glucose molecule oxidized. Approximately 40% of the released energy is
captured to make this ATP, while the rest is lost as heat. This is a highly efficient process compared to the process of
fermentation which is an anaerobic process (that is, one that occurs in the absence of oxygen). Fermentation yields a net of only
2 molecules of ATP per glucose.
A byproduct of cellular respiration is carbon dioxide. CO2 generated by human cells diffuses into the blood stream where it is
taken to the lungs. Note from the previous chemical equation that the amount of CO 2 generated is equal to the amount of oxygen
consumed (assuming that the source of energy is glucose). Therefore, if a metabolically active plant or animal is confined in a
closed chamber and the CO2 generated is removed from the chamber, the volume of air in the chamber will decrease in proportion
to the amount of O2 used for cellular respiration.

Activity A –Plant Cellular Respiration Rate Assay: Influence of Temperature

and Germination Status
The plant respiration assay measures the utilization of oxygen by seeds in a special container. The class will investigate the effects
of temperature on aerobic respiration in germinating and non-germinating seeds. Each group of 3-4 students will need four set-
ups:
(1) Approximately 12 germinating seeds at 3 to 5C (ice-water bath)
(2) Approximately 12 germinating seeds at room temperature (22 – 24C)
(3) Approximately 12 germinating seeds at 45 C (warm-water bath)
(4) Approximately 20 non-germinating seeds at room temperature (22 – 24C)

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Procedure:

1. Place approximately 12 germinating seeds in each of three weighing dishes. Use forceps to
select the seeds and be careful not to damage them during the transfer. Do not select seeds
with a substantial amount of mold on them.

2. Place approximately 20 non-germinating seeds in a weighing dish.

3. Records the weight of each weighing dishes together with the seeds it contains. Subtract the
approximate weight of a weighing dish to determine the weight of each of the seed samples.
Record these weights in the appropriate data table in your report.

4. Place each sample into an individually –labeled large test tube (respiration chambers). Make
certain you record the respective test tube number/temperature condition into which each of
the weighed seed samples was placed.

5. Place a loose wad of cotton over the seeds in the tube, and use a spatula to gently push the
cotton down into the tube (see the diagram below)

Graduated tube

Soda lime

Cotton plug

Seeds

The cotton should not be packed so tightly as to interfere with gas flow. The “depth” of the
cotton layer should be no more than ½ inch (approximately 1cm). The cotton keeps the soda
lime (which will be added in the next step) from damaging the seeds.

6. Using a spatula, add about 1 cm of soda lime (which is sodium hydroxide and lime) over the
cotton. Caution: Do not touch the soda lime. If it does come into contact with your skin,
wash your hands with soap and water. The soda lime will function to remove the expired
carbon dioxide from the tube.

7. Firmly insert the rubber stopper with the attached graduated pipette into each of the four test
tubes. Note the measurement scale on the graduated tube.

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8. Place the “3-5C” tube in a test tube rack in the ice-bath. Place the “45C” tube in a test tube
rack in the hot water-bath. Leave both of the “22-24C” tubes (germinating and non-
germinating seeds) in a test tube rack on your bench at room temperature.

9. Allow all respiration tubes to “equilibrate” to their respective experimental temperatures for a
minimum of 5 minutes.

10. Using a tadpole pipette, add enough colored water (enough to give a band approximately 1/8
in. long) to the end of each of the four graduated pipettes. Make certain that approximately the
same volume of dye is in all four graduated pipettes. The pipettes are marked in increments
from 0 to 1 ml. For each test tube, immediately record the position (in ml) of the front of
the colored dye band, and record the starting time, in the data table in your report.

11. Continue the experiment until the dye band in any one of the four tubes has moved through
most of the graduated pipette. At this point, check each tube, and record both the final
position of the dye band (in ml), and the stop time.

12. By comparing the starting position of the dye with the final position, determine the volume of
oxygen used during the experiment for each tube.

13. Calculate the volume of oxygen consumed per minute per gram of seeds (ml/min./g) for each
tube, and record the results in the data table. (12 and 13 are done in the last table).

At the conclusion of the experiment, remove the stoppers from the test tubes and rinse the colored water out of the
graduate pipettes. Discard the soda lime in designated containers!! Discard the cotton balls in the trash can. Remove
the seeds and return them to their proper places (Do NOT throw them in the trash can!).
Rinse the test tubes out and return them to the rack on your table. Make certain you wash your hands with soap and
water if you handled the soda lime.

Activity B – Fermentation

Fermentation is another way that some cells can produce ATP by oxidizing organic molecules.
Fermentation is far less efficient, however, than cellular respiration. Fermentation takes place in
the absence of oxygen, and is thus an anaerobic process. Some cells (such as yeasts and some
types of bacteria) produce ethyl alcohol as a byproduct of fermentation. Other cells produce
lactic acid during fermentation (for example, muscle cells when oxygen becomes depleted, or the
bacterial cultures used to produce yogurt).

In this activity, you will mix yeast with corn syrup (a source of the sugars sucrose and fructose)
under conditions where oxygen is excluded. The yeast contain the enzymes and other cellular
machinery needed to process these sugars and produce alcohol by fermentation. You will
measure the rate of fermentation by measuring the production of carbon dioxide (CO 2), another
product of fermentation.

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Procedure:

1. Get three large test tubes and label them 1, 2, and 3. Mix solutions of yeast suspension, 50%
corn syrup, and water in these tubes according to the following table.

Tube Number (volumes shown in ml)

1 2 3
Water 6 3 0
Yeast suspension 0 3 6
Corn syrup (50%) 12 12 12

2. Mix the contents of each tube thoroughly using a glass stirring rod, and transfer the contents to
a clean fermentation tube (“turkey tube”) that has been marked with the same number as the
respective test tube. Tip the fermentation tube back to remove any air from the upright part of
the tube, then add more solution to the bulb of the tube. The start and end times for each of
your three tubes has been recorded.

3. Carbon dioxide gas will begin to build up in the upright column of the turkey tube. Measure the
height of the column of carbon dioxide gas at five minute intervals, and record the results in the
data table provided. Continue recording data for at least 20 minutes.

Make a line graph of your results, showing the amount of carbon dioxide produced (mm) as a
function of time (minutes) for your three different reactions. The data for this graph is located in
the table below this paragraph. Use different lines for the data for tube #1, tube #2, and tube
#3. http://onlinecharttool.com/graph?selected_graph=line

Height of carbon dioxide column in millimeters (mm):

Tube number Minutes
0 5 10 15 20 25 30
1 (0 ml yeast) 0 0 0 0 0 0 0

2 (3 ml yeast) 1 2 3 4 5 6 7

3 (6 ml yeast) 2 4 6 8 10 12 14

Report Sheet
Cellular Respiration Lab
Submit the statement and questions with your answers.
Always include the number and statement/question with your answers.

Evaluate each of the following statements and decide if it is “True” (T) or “False” (F).

1.____ Oxygen is required for aerobic respiration.

2.____ Carbon dioxide is considered a product of the chemical reaction for cellular respiration.

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3.____ During cellular respiration, glucose is oxidized by combining with oxygen.

4.____ The net ATP yield from fermentation is greater than from aerobic respiration.

5.____ An assumption made in our cellular respiration experiment is that the volume of air in the
closed chamber will decrease proportionately to the amount of oxygen used for cellular
respiration.

6.____ The purpose for the soda ash used in our cellular respiration experiment is to keep the air
in the closed chamber at approximately pH 7.

8. What are your conclusions regarding the effects of temperature on the rate of respiration of
germinating pea seeds? Also, explain what you conclude about cellular respiration in non-
germinating seeds.

9. What is the independent variable in Activity A?

10. What is the dependent variable in Activity A?

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Calculate the Volume of air consumed, Oxygen/minute, and Oxygen/minute/gram.

Temp. Seed Seed Start time Stop time Vol. of Air Number of O2/min. O2/min/gram
(C) Condition Weight (min:sec) (min:sec) consumed Minutes
(grams) (ml) elapsed
22-24 non-germ. 20 0 20” 0 20”
22-24 germinating 25 0 20” 3 20”
3-5 germinating 21 0 20” 0 20”
45 germinating 20 0 20” .5 20”

http://www.scribd.com/doc/14611990/AP-Biology-Cellular-Respiration-Lab-Report

http://www.biologycorner.com/worksheets/cellular_respiration_virtual.html

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