9477 / 2027

Lesson 5 of 5 / Respiration and ATP production

Measure respiration fairly

What does a moving respirometer liquid actually measure?

In this lesson: Investigate temperature, oxygen and substrate effects on respiration.

About 7 min

The key ideaA valid respiration measurement separates biological gas exchange from temperature, pressure and sample-size effects.

Explore the idea

Separate oxygen uptake from physical gas changes

Marker movestowards vesselSeeds take up O2CO2 absorbent,separated from seedsControlled water-bath temperature

The living sample shows 1.00 cm3 apparent gas uptake. The marker alone cannot distinguish respiration from physical gas changes. Inspect the matched non-respiring control before calculating the biological rate.

These are calculation scenarios, not a model predicting how a real sample changes. Capillary volume change = displacement x cross-sectional area; correction must preserve movement direction. Use comparable displaced volume in the control, allow temperature equilibration, check seals and repeat measurements. The simplified apparatus omits connections used to equalise pressure before readings. Practise laboratory setup and reagent safety with supervision.

Explanation

A simple respirometer can measure oxygen uptake by respiring material. If carbon dioxide is absorbed by a suitable reagent, a fall in gas volume or pressure mainly reflects oxygen removal. A marker in a capillary then moves, allowing volume change to be estimated from distance and cross-sectional area.

Temperature and atmospheric pressure can also move gas or liquid. Use a matched control with non-respiring material of comparable displaced volume and allow equilibration in a water bath. Keep the apparatus sealed, compare equal living mass or normalise rate to mass, and repeat independent measurements.

Substrate supply and oxygen availability can limit respiration, while temperature changes enzyme activity and can eventually damage cells. Changing one factor while controlling others is essential. In intact organisms, uptake and transport of substrate can also matter, so a response is not automatically the direct effect on one enzyme.

For yeast under defined oxygen-limited conditions, CO2 or ethanol production can indicate fermentation rate. It is not directly comparable to oxygen uptake without specifying the pathway. Use suitable supervised laboratory materials and risk controls for caustic CO2 absorbents, hot water and pressure; a lesson model is not a substitute for laboratory training.

Step by step
  1. 1

    Define the biological signal

    Oxygen uptake or fermentation product formation?

  2. 2

    Remove physical confounds

    Control temperature, pressure and leaks.

  3. 3

    Calculate a comparable rate

    Use volume per time per mass when appropriate.

Worked example

Work through the evidence

After correction with a control, a 2 g seed sample takes up 0.8 cm3 O2 in 4 minutes. Calculate rate per gram.

One way to explain it

0.8/(4 x 2) = 0.10 cm3 O2 min-1 g-1.

Why this answer works
  • Divide by time and living mass.
  • Use the corrected gas change, not an uncorrected marker displacement.
Is this true? "Every gas-volume fall proves oxygen consumption by living cells."

Cooling, pressure changes and leaks can also affect gas volume; controls are needed.

Try a question

Why include a matched non-respiring control?
You can return to this lesson any time.