Mr. Brown · Biology · Lab Investigation

Floating Leaf Disks

A Photosynthesis Assay

Welcome. Today you will use a clever, low-tech assay to measure how fast a leaf can photosynthesize — by watching disks rise from the bottom of a beaker. You'll manipulate variables, gather data, and reason about why one disk takes longer than another.

⏱ Estimated time: 30 minutes. Your work auto-saves silently to this device.

Part 1 · Reading with Vocabulary

🌿 What's Going On Inside the Leaf?

A leaf is a chemical factory. Inside every green leaf, microscopic organelles called chloroplasts capture light and use that energy to split water and stitch carbon dioxide into sugar. The pigment that absorbs the light is called chlorophyll, and it absorbs red and blue wavelengths much more than green — which is exactly why a leaf looks green to your eye. The leftover green light is the light the leaf didn't use.

The reaction we call photosynthesis takes in CO₂ and water and outputs glucose plus oxygen gas. A leaf normally exchanges these gases through tiny pores on the underside of the leaf called stomata. The spongy interior — the mesophyll — is full of air spaces. That trapped air is the reason a leaf disk floats: it's not denser than water, it's just full of bubbles.

If we replace that internal air with liquid (using a syringe and a little suction — vacuum infiltration), the disk sinks. Now we have a trick: as the leaf photosynthesizes, it releases oxygen. The O₂ fills the air spaces back up, and the disk rises again. Because the leaf is also doing cellular respiration at the same time (consuming O₂), what we actually measure is net photosynthesis — the photosynthesis happening on top of the respiration. Gross photosynthesis is the total rate before respiration is subtracted off.

To make sure the leaf has enough CO₂ to work with, we infiltrate with a bicarbonate solution (NaHCO₃), which acts as a CO₂ source dissolved in the water. The faster the leaf can build sugar, the faster O₂ accumulates, the sooner the disk's buoyancy wins out and it rises. We summarize a whole trial with one number: ET50, the time when half of the disks have floated. A shorter ET50 means a faster rate.

Several factors can change the rate: how bright the light is (light intensity), what color it is (wavelength), how much CO₂ is dissolved, and the temperature of the water. Today you'll change one factor at a time and ask, "Did the disks rise faster, slower, or the same?"

Big idea: The disks aren't producing oxygen because they're floating — they're floating because they're producing oxygen.
Part 1 (cont.) · Vocabulary Cards

📇 Vocabulary — One Card at a Time

Tap a card to flip it. Only one card opens at a time. Each open card stays revealed for 8 seconds before re-closing. You can open the same card more than once.

Once you've reviewed every card at least once, the Continue button activates and takes you to the matching exercise.
Part 1 (cont.) · Matching Practice

🔗 Match the Term to the Meaning

Click a term on the left, then click its definition on the right. Correct matches dim out. Each correct match = 1 pt.

Terms

Definitions

Score: 0Mismatches shake but don't count against you.
Part 2 · Pre-Lab Thinking

🧠 Pre-Lab Questions

Short responses. Each question = 1 pt for an answer that addresses the question.

Q1. Why does an untreated leaf disk float on water before any infiltration step?
Q2. How does vacuum infiltration cause the disks to sink?
Q3. Why is sodium bicarbonate (NaHCO₃) added to the infiltration solution instead of using plain water?
Q4. How would you keep this an experiment about light intensity rather than light and temperature?
Q5. ET50 is defined as the time when 50% of the disks have floated. Explain why a shorter ET50 means a faster rate of net photosynthesis.
Q6. A student claims their disks are floating because the water is heating up under the lamp and the disks are expanding. Argue against this claim using what you know about gas exchange in the leaf.
Part 3 · Simulation

🧪 Run the Floating Disk Simulation

What you're looking at. Below is a virtual beaker holding 10 leaf disks that have already been vacuum-infiltrated with bicarbonate solution. They sit at the bottom. When you press START, the simulated lamp turns on and the disks rise at a rate determined by your settings. The clock measures the time, in seconds, until each disk floats. The simulation reports the ET50 — the time when 5 of the 10 disks have surfaced. Use this number as your rate proxy.

Directions. Run at least 4 trials: one trial at each light intensity (Low, Medium, High) with the white filter, then one extra trial of your choice. Record results in the data table on the next page. The simulator stops automatically at 240 seconds if not all disks have surfaced — record "NF" (did not float) if so.

Time: 0.0 s Floated: 0 / 10 ET50:

Simulation Observation Questions

Q7. Run the trial at Low light (white filter, 22 °C). What is the ET50?
Q8. Run again at High light. What is the ET50? How does it compare to Low light?
Q9. Now switch the filter to green with the same intensity you used in Q8. Predict first, then test. Were you right? Why did the rate change as it did?
Q10. Set the temperature to 38 °C with high light and white filter. Does the rate keep increasing, or does it slow back down? Why would that happen biologically?
Part 4 · Data

📊 Record Your Trials

Fill in all 8 trials below. A complete table is worth 4 pts (full credit if every cell has a value).

TrialLight IntensityFilter / ColorTemp (°C)ET50 (s)Rate (1/ET50)
1
2
3
4
5
6
7
8
Rate calculation reminder: Rate = 1/ET50. If your ET50 is 80 s, your rate is 1/80 = 0.0125 disks/second. A larger rate = faster photosynthesis.
Table: 0 / 4
Part 5 · Graph & Data Analysis

📈 Reading a Class Dataset

The graph below shows class-averaged rates (1/ET50) versus light intensity at 22 °C. Use it to answer the questions.

0.025 0.020 0.015 0.010 0.005 0 100 300 600 1000 1500 Light intensity (μmol photons · m⁻² · s⁻¹) Rate (1/ET50, s⁻¹) Light-Response Curve · Spinach Disks · 22 °C · White Light light saturation
Q11. At what approximate light intensity does the curve plateau (the leaf becomes light-saturated)?
Q12. Estimate the rate at 300 μmol m⁻² s⁻¹ from the curve.
Q13. The curve never reaches zero at the lowest light shown. Explain why even at low light there is still some net photosynthesis.
Q14. A different class measures spinach at 22 °C, but their curve plateaus at a lower rate (about 0.012). Propose two factors that could explain a lower plateau than the one shown.
Q15. What would happen to this curve if the CO₂ supply were reduced? Describe the shape change.
Part 6 · Case Study

📁 Case Study: Coastal Ivy in a Warming Tide Pool

Background

Dr. Mireille Okafor studies salt-tolerant coastal plants at a research station in northern California. In summer 2025, she noticed that the ivy growing closest to a shallow tide pool — which warms to around 32 °C on still afternoons — appeared paler and grew more slowly than ivy 10 m inland. She wondered whether photosynthesis itself was being affected by the warmer microclimate, or whether the salt spray was the real culprit.

To test this, she ran a floating-leaf-disk assay using ivy from three locations: (A) next to the warm tide pool, (B) 10 m inland in moderate shade at ~22 °C, and (C) 10 m inland in full sun at ~28 °C. All trials used 0.2% sodium bicarbonate solution, identical LED lighting at 800 μmol m⁻² s⁻¹, and the assay was run at 22 °C laboratory temperature for fairness. Results from 30 disks per group:

GroupSourceET50 (s)Notes
ATide pool (32 °C native)178Pale, smaller disks
BInland shade (22 °C native)96Dark green, supple
CInland sun (28 °C native)71Thicker, waxy disks

Dr. Okafor also recorded that the ivy at the tide pool was not visibly salt-burned. A quick chlorophyll extraction showed Group A had roughly 40% less chlorophyll per gram than Group B.

Case Study Questions

Q16. Rank the three groups from fastest to slowest net photosynthesis rate based on the data. Justify your ranking in one sentence.
Q17. The lab temperature was held at 22 °C for all groups even though the leaves came from different native temperatures. Why was this an important design choice?
Q18. Group A has 40% less chlorophyll per gram than Group B. Explain mechanistically how a lower chlorophyll content would slow the floating rate, even if the leaf disks are the same size.
Q19. Dr. Okafor's first hypothesis was that heat itself was damaging the tide-pool ivy. Does this experiment support that hypothesis? Why or why not? Be specific about what the assay does and doesn't tell us.
Q20. Design a follow-up experiment using the floating disk assay to determine whether the slow rate in Group A is caused by chronic heat exposure, salt spray, or something else. Identify the independent variable and at least two controls.
Q21. Sun-grown leaves (Group C) photosynthesized fastest. Why might this be evolutionarily/physiologically expected for plants grown in high-light conditions?
Part 7 · Free Response

✍️ Free Response Question

Answer all parts. Each lettered part = 1 pt.

Scenario

A biology student is investigating how the wavelength of light affects the rate of photosynthesis in spinach leaves using the floating-leaf-disk assay. They hold light intensity at 800 μmol m⁻² s⁻¹, temperature at 22 °C, and use 0.2% sodium bicarbonate. They run trials under four colored filters: red, blue, green, and a no-filter (white-light) control. Each trial uses 10 fresh disks.

Q22a. State a hypothesis for which color filter will produce the shortest ET50 and which will produce the longest. Justify your hypothesis using what you know about chlorophyll absorption.
Q22b. Identify the independent variable, dependent variable, and at least three controlled variables in this design.
Q22c. The student gets these ET50 values: White = 68 s, Red = 79 s, Blue = 84 s, Green = 198 s. Calculate the rate (1/ET50) for each and state which filter gave the slowest rate.
Q22d. A classmate argues that "green light doesn't work for photosynthesis at all" based on these results. Use the data to refute this stronger claim and clarify what the data actually shows.
Q22e. Propose one source of error in this experiment and one specific modification that would reduce it.
Part 8 · Mastery Quiz

🎯 Mastery Quiz (10 Questions)

Questions are drawn at random from a large bank, so your set is different from your neighbor's. Each correct answer = 1 pt. You get 2 attempts; your higher score counts.

Attempt 1 of 2 0 / 10
Part 9 · Final Grade

🏁 Final Score

— / —
Letter grade scale: 90+ = A · 80+ = B · 70+ = C · 60+ = D · below 60 = F

Reflection (not graded)

What surprised you most about how light, color, CO₂, or temperature affected the leaf disks?

Mr. Brown · Biology · Floating Leaf Disk Photosynthesis Assay · v1.0