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Study Photosynthesis with the Floating Leaf Disk Assay

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Grade Range
Group Size
3 students
Active Time
Part 1: 1 h 40 minutes, Part 2: 1 hour 45 minutes
Total Time
Part 1: 1 h 40 minutes, Part 2: 1 hour 45 minutes
Area of Science
Plant Biology
Key Concepts
Svenja Lohner, PhD, Science Buddies
Cup filled with a solution and leaf disks on the bottom. One leaf disk is floating to the surface.


Plants carry out photosynthesis to produce sugars that they need as an energy source to live and grow. During photosynthesis, oxygen—a gas that many living beings need to survive—is released. This makes photosynthesis one of the most important biological processes on Earth. In Part 1 of this lesson plan, students will utilize the floating leaf disk assay to demonstrate the production of oxygen gas during photosynthesis. They will then continue to design and conduct their own experiments in Part 2 of the lesson in order to investigate variables that affect the rate of photosynthesis in plants.

Remote learning adaptation: This lesson plan can be conducted remotely. The Engage section of the lesson can be skipped or done over a video call, then students can work independently during the Explore sections, using the Student Worksheet and the Leaf Disk Assay Video as a guide. Students will need to obtain their own materials such as a light source and a plastic syringe. For Part 2 of the lesson, students can adjust the independent variable they will investigate according to the materials available to them. The data analysis in the Reflect sections and the presentations of the results can be done over a video call, or by sharing a poster or slides on a class drive.

Learning Objectives

NGSS Alignment

This lesson helps students prepare for these Next Generation Science Standards Performance Expectations:
This lesson focuses on these aspects of NGSS Three Dimensional Learning:

Science & Engineering Practices Disciplinary Core Ideas Crosscutting Concepts
Science & Engineering Practices Planning and Carrying Out Investigations. Plan an investigation individually and collaboratively, and in the design: identify independent and dependent variables and controls, what tools are needed to do the gathering, how measurements will be recorded, and how many data are needed to support a claim.

Conduct an investigation and/or evaluate and/or revise the experimental design to produce data to serve as the basis for evidence that meet the goals of the investigation.

Collect data about the performance of a proposed object, tool, process, or system under a range of conditions.

Analyzing and Interpreting Data. Analyze and interpret data to determine similarities and differences in findings

Engaging in Argument from Evidence. Construct, use, and/or present an oral and written argument supported by empirical evidence and scientific reasoning to support or refute an explanation or a model for a phenomenon or a solution to a problem.
Disciplinary Core Ideas LS1.C: Organization for Matter and Energy Flow in Organisms. Plants, algae (including phytoplankton), and many microorganisms use the energy from light to make sugars (food) from carbon dioxide from the atmosphere and water through the process of photosynthesis, which also releases oxygen. These sugars can be used immediately or stored for growth or later use.
Crosscutting Concepts Energy and Matter. Within a natural system, the transfer of energy drives the motion and/or cycling of matter.


Materials needed for the 'Study Photosynthesis with the Floating Leaf Disk Assay' lesson.

For each student group of 3:

For teacher:

Extra materials for independent student investigations:

Background Information for Teachers

This section contains a quick review for teachers of the science and concepts covered in this lesson.

Every living organism needs energy to survive, to grow, and to reproduce. Humans and animals eat foods with carbohydrates, proteins, and fats to produce the energy they need to survive. But plants do not eat. They make their own energy source in the form of energy-rich carbohydrates (sugars) through a process called photosynthesis. Photosynthesis is a multi-step, enzyme-mediated process that converts light energy into chemical energy. During photosynthesis, plant cells use light energy (such as light emitted from the Sun), water (H2O), and carbon dioxide (CO2) as reactants to produce sugar molecules (C6H12O6) and oxygen (O2) (Figure 1):

 A schematic drawing of a plant showing all the plant parts above and underground. In the soil, arrows labeled H2O point to the plant roots. Yellow arrows coming from the top indicate sunlight shining onto the plant. An arrow labeled CO2 is pointing toward the plant leaves. An arrow labeled O2 and another arrow labeled C6H12O6  are pointing away from the plant leaves.
Figure 1. During photosynthesis, plants convert water (H2O), carbon dioxide (CO2), and light into oxygen (O2) and sugars like glucose (C6H12O6).

Photosynthesis takes place in the chloroplasts within the plant's cells. The chloroplasts contain special pigments that react to light. Chlorophyll is one of the pigments that can absorb light in the blue and red spectrum from the visible light spectrum. Chlorophyll does not absorb light in the green spectrum of light, but reflects it instead. This is why leaves with chlorophyll usually appear green. During the first part of photosynthesis—the light-dependent reaction—chlorophyll and other pigments harness the light energy to produce NADPH and ATP, which are two types of energy-carrier molecules. At the same time, water is split into oxygen (O2) and protons (H+). The next stage is light-independent and is often referred to as the dark reaction. In this step, the two energy-carrier molecules NADPH and ATP are utilized in a series of chemical reactions called the Calvin cycle. In the Calvin cycle, the plants take carbon dioxide (CO2) from the air and use it to ultimately make sugars such as glucose or sucrose. These sugars can be stored for later use by the plant as an energy source to fuel its metabolism and growth.

Photosynthesis is responsible for replenishing Earth's atmosphere with oxygen that we breathe. Thus, it is not only crucial for plants, but also for all organisms that rely on oxygen for their survival. Many factors affect how quickly plants are able to conduct photosynthesis. Without enough light or water, for example, a plant cannot photosynthesize very quickly. Similarly, the concentration of carbon dioxide—another reactant in photosynthesis—affects how fast photosynthesis can occur. Temperature also plays a significant role, as photosynthesis is an enzyme-mediated reaction. This is because at high temperatures, enzymes can get damaged and thus become inactivated. Other factors that affect the rate of photosynthesis are the light intensity, the amount of chlorophyll or other color pigments in a plant, and the color of light.

Similar to any other chemical reaction, the rate of photosynthesis can be determined by either measuring the decrease of its reactants or the increase of its products. You could, for example, measure the consumption of carbon dioxide or the production of oxygen over time. Without the use of extensive laboratory equipment, the rate of photosynthesis can be determined indirectly by conducting a floating leaf disk assay to measure the rate of oxygen production (Figure 2). In the floating leaf disk assay, 10 or more leaf disk samples are punched out of a leaf. In the next step, a vacuum is used to replace the air pockets within the leaf structure with a baking soda (bicarbonate) solution. The dissolved baking soda provides the carbon dioxide that the leaf needs for photosynthesis. The leaf disks are then sunk in the baking soda solution and exposed to light. As the plant leaf photosynthesizes, oxygen is produced that accumulates as oxygen gas bubbles at the outside of the leaf disk. The attached oxygen gas changes the buoyancy of the leaf disk and once enough oxygen has been produced the leaf disk will rise to the surface of the baking soda solution. The time from exposure to light until the leaf disk rises to the top of the solution is a measure of how much oxygen has been produced and thus a proxy for the rate of photosynthesis.

 Cup filled with a solution and leaf disks on the bottom. One leaf disk is floating to the surface.
Figure 2. The floating leaf disk assay allows to indirectly determine the rate of photosynthesis.

In this lesson plan, students will place 10 disks in the baking soda solution at the same time. A good way to collect data is to count the number of floating disks at the end of a fixed time interval; for example, after every minute until all disks are floating. The time required for 50% of the leaves to float represents the Effective Time (ET50). ET50 can be determined by timing when the fifth leaf floats, or by graphing the number of disks floating over time, as shown in Figure 3. An ET50 of 11.5 minutes (min), for example, as shown in Figure 3, would mean that after 11.5 min 50% of the leaves (5 out of the 10) floated on top of the baking soda solution. In the context of oxygen production, you could also say that an ET50 value of 11.5 min means that it took 11.5 min to produce enough oxygen to make 50% of the leaf disks float.

 A scatter plot graph showing exemplary results for a leaf disk assay.

The x-axis shows time in minutes. The y-axis shows the number of floating leaf disks. After 7 minutes the first leaf disk floats, after 11 minutes 4 leaf disks float, at 12 minutes 7 leaf disks float, at 13 minutes 8 leaf disks float, and after 14 minutes all 10 leaf disks float. A red line indicates at what time 50% (5) leaf disks float (at about 11.5 minutes). This time is labeled Effective Time ET50.

Figure 3. Example results for the floating leaf disk assay. The graph shows the time on the x-axis and the number of floating leaves on the y-axis. The Effective Time (ET50) represents the time required for 50% of the leaves to float. By extrapolating from the graph, the 50% floating point in this graph is about 11.5 min.

Reaction rates are usually expressed as the concentration of reactant consumed or the concentration of product formed per unit of time. As mentioned above, we can use the ET50 as a proxy for how much oxygen has been produced to make half of the leaf disks float. This means that the ET50 value is proportional to the inverse of the rate of oxygen production, or proportional to the inverse of the rate of photosynthesis. The reciprocal of ET50 or 1/ET50 can thus be used as a simple measure of the rate of photosynthesis.

This lesson plan has two parts. In the first part, students use the leaf disk assay to explore how plants make energy using photosynthesis. In the second part, students design and conduct experiments that use the leaf disk assay to investigate several variables that have the potential to affect the rate of photosynthesis. This allows students to apply and review scientific concepts involved in photosynthesis, such as cell structure and function, enzyme activity, energy use and storage, and reaction rates.

Prep Work (15 minutes)

Engage (10 minutes)

Explore (45 minutes)

Reflect (45 minutes)


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