elodea experiment

Practical Biology

A collection of experiments that demonstrate biological concepts and processes.

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Practical Work for Learning

Published experiments

Investigating factors affecting the rate of photosynthesis, class practical.

In this experiment the rate of photosynthesis is measured by counting the number of bubbles rising from the cut end of a piece of Elodea or Cabomba .

Lesson organisation

The work could be carried out individually or in groups of up to 3 students (counter, timekeeper and scribe).

Apparatus and Chemicals

Students may choose to use:.

Thermometer, –10 °C –110°C

Coloured filters or light bulbs

Push-button counter

Potassium hydrogencarbonate powder or solution (Hazcard 95C describes this as low hazard)

For each group of students:

Student sheets, 1 per student

Beaker, 600 cm 3 , 1

Metre ruler, 1

Elodea ( Note 1 ) or other oxygenating pond plant ( Note 2 )

Electric lamp

Clamp stand with boss and clamp

Health & Safety and Technical notes

Normal laboratory safety procedures should be followed. There is a slight risk of infection from pond water, so take sensible hygiene precautions, cover cuts and wash hands thoroughly after the work is complete.

Read our standard health & safety guidance

1 Elodea can be stored in a fish tank on a windowsill, in the laboratory or prep room. However it is probably a good idea to replace it every so often with a fresh supply from an aquarist centre or a pond. (It’s worth finding out if any colleague has a pond.) On the day of the experiment, cut 10 cm lengths of Elodea , put a paper-clip on one end to weigh them down and place in a boiling tube of water in a boiling tube rack, near a high intensity lamp, such as a halogen lamp or a fluorescent striplight. Check the Elodea to see if it is bubbling. Sometimes cutting 2–3 mm off the end of the Elodea will induce bubbling from the cut end or change the size of the bubbles being produced.

2 Cabomba (available from pet shops or suppliers of aquaria – used as an oxygenator in tropical fish tanks) can be used as an alternative to Elodea , and some people find it produces more bubbles. It does, though tend to break apart very easily, and fish may eat it very quickly.

3 If possible, provide cardboard to allow students to shield their experiment from other lights in the room.

Ethical issues

Look out for small aquatic invertebrates attached to the pond weed used, and remove them to a pond or aquarium.

Activities Associated with this Lesson Units serve as guides to a particular content or subject area. Nested under units are lessons (in purple) and hands-on activities (in blue). Note that not all lessons and activities will exist under a unit, and instead may exist as "standalone" curriculum.

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Students perform data analysis and reverse engineering to understand how photosynthesis works. Both are important aspects of being an engineer.

After this activity, students should be able to:

• Explain that photosynthesis is a process that plants use to convert light energy into glucose, a source of stored chemical energy for the plant.
• Describe photosynthesis as a set of chemical reactions in which the plant uses carbon dioxide and water to form glucose and oxygen.
• Describe a simple experiment that provides indirect evidence that photosynthesis is occurring.
• Describe the effects of varying light intensity on the amount of photosynthesis that occurs.

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• 5 liters (about 1¼ gallons) of aged tap water (tap water in an open container that has been allowed to sit for 36-48 hours to eliminate the chlorine used in municipal water supplies)
• 15-20 total Elodea plants; these are hardy freshwater aquarium plants sold in bunches at pet stores and suppliers such as Carolina Biological Supply Company (www.carolina.com)
• string, yarn or twist ties for tying Elodea plants into bunches
• small rocks or similar objects to serve as weights to hold the Elodea plants underwater
• 500-ml beakers, 1 per team
• baking soda, a few tablespoons (sodium bicarbonate)
• timers or watches with second hands, 1 per team
• small adjustable desk lamps that can be set up so that their light bulbs are a few inches above the beakers and shine vertically down onto them; flashlights with strong beams that are mounted on ring stands also work; 1 light source per team

An understanding of photosynthesis, as presented in the associated lesson, Do Plants Eat?

(Get the class' attention and ask them to do as you say.) With one hand, pinch your nose closed. Raise your other hand high in the air. Now take a deep breath and hold it for as long as you can. When you cannot hold your breath any longer, lower your raised hand and unpinch your nose. (Once all hands are down and no one is left holding their breath, move on.) Why did you need to start breathing again? (From their elementary school studies, expect students to be able to tell you that their bodies need air in order to survive.)

What, exactly, is in air? (Students may not know that air contains more than oxygen.) Most of the air we breathe—the atmosphere—consists of nitrogen gas (about 78%). Oxygen is the next largest component (about 21%) and a tiny part (1%) is made up of argon (an inert gas), water vapor and carbon dioxide.

So, specifically what component(s) of air do our bodies need? (Expect them to be able to answer that it is oxygen.) And what do our bodies do with oxygen? That's right, oygen from the air is picked up in the lungs by the blood and carried to all parts of the body, where it is used by muscles and the brain and all the other organs and tissues of the body. We cannot live without it.

From where did the oxygen in the atmosphere come? (They may know or be able to reason that it is the result of all the plants that have lived on the Earth and have been doing photosynthesis for many millions of years.) Today, you will work in teams to conduct an experiment to see if the amount of light plants receive can affect this production of oxygen.

Overall Experiment Plan

• In a class discussion format, students establish a hypothesis to be tested by the class in the experiment.
• Working in teams, students set up and conduct the experiment. Each team conducts two trials: one with the plants lit only by the ambient light available in the classroom when some or all of the room lights are turned off, and one with the plants receiving bright light from the desk lamps. The data collected are the number of bubbles of oxygen that are given off by the plants in a five-minute period, first at low-light levels, and then at high-light levels.
• Then the groups come together to pool their data from each of the two trials. From these data, students individually determine the mean, median and modes for the numbers of bubbles produced during the two different light conditions.
• Then students individually graph the data, using bar graphs that show the mean numbers of bubbles and the ranges for each test condition.

Part 1: Generating a Hypothesis

Explain to the class that before researchers start experiments, they first create a prediction about the expected outcome of the experiment. This prediction is known as a hypothesis. A hypothesis is not simply a guess, however. Instead, it is a prediction based on prior knowledge of or experience with the subject. For example, if a gardener wanted to find out if it was really necessary to fertilize zucchini plants, they might grow 12 zucchini plants, but fertilize only half of them. In this case, the hypothesis being tested might be: Fertilized zucchini plants produce more zucchinis than unfertilized zucchini plants. The data collected to support or refute the hypothesis would be the total number of zucchinis produced by the fertilized plants, compared to the total number produced by the unfertilized plants.

Point out that in the zucchini experiment, the gardener collected data that involved numbers. In science, this is usually the case, because numbers can easily be compared and are cumulative for many things that actually happen, as opposed to things that the experimenter thought might happen.

Then, explain briefly how the photosynthesis experiment will be set up and ask the class to determine a hypothesis to be tested. It shouldn't take them long to come up with a statement such as: The plants that receive more light produce more bubbles than the plants that receive less light.

Part 2: Setting up the Experiment

Perform the following steps with some or all of the classroom lights turned off. Ideally, the room should not be brightly lit, nor should it be dark; adequate light should be present for students to easily see.

• Each team fills a beaker with about 500 ml of aged water for the Elodea. To this water, add a scant one-quarter teaspoon of sodium bicarbonate (baking soda) to provide a source of carbon dioxide for the plants, since they cannot get it from the atmosphere like terrestrial plants do. Stir the water until the sodium bicarbonate is dissolved and the water looks clear.
• Each team obtains enough sections of Elodea plants so that it has about 18-24 inches of total plant length. Arrange them so that all of the plants are at least 1½" under the water in the beaker. Use string or twist ties to hold them together, and then add a small rock to keep the plants from floating to the surface. Point out that the more area exposed to the light above the plant, the more photosynthesis can occur within the leaves. If students form clumps of Elodea, many of leaves will be shaded by those above, and thus may not be able to perform as much photosynthesis. It is best to form the plants into loops that cover the entire bottom of a beaker, instead of a single clump in the middle of the beaker.

Part 3: Running the Experiment

• As soon as the plants are arranged in the beakers, have the team start timing for five minutes. Direct two team members to have their eyes glued to the beaker for those five minutes, watching for bubbles to rise to the water surface. Announce to the third team member the sighting of any bubbles that rise, so s/he can keep count (using tally marks is helpful) and monitor the time, indicating when the five minutes are up. The bubbles are fairly large, about 2 mm in diameter, and so are easily seen when they rise to the surface.
• When all teams have counted bubbles for five minutes (it is quite possible that some teams see no bubbles at all), turn on the room lights and have students position the desk lamps directly above the beakers with the light bulbs only be a few inches above the beakers. Once the lights are in place, have the teams again begin timing and counting/recording bubbles for five minutes.

Part 4: Pooling and Analyzing the Data

• Make a large chart on the classroom board in which teams can fill in the number of bubbles they counted during each of the two light conditions.
• Once the chart is filled in, have students work individually to determine the mean, median, mode and range of each of the two data sets. Allow enough time so that all students arrive at the same answers.
• Provide students with grid paper and direct them to make vertical bar graphs that compare the mean number of bubbles in the two light conditions. Be sure that students include titles, axes labels and legends if different colors are used for the two bars. Then show them how they can indicate the ranges of the data by adding a vertical line segment to the center top of each bar, with the lower end of the line segment situated at the lowest number of bubbles observed by a team, and the upper end of the line segment at the highest number of bubbles observed.

Part 5: Interpreting the Data

• As a class, examine all the data and graphs and revisit the hypothesis. What do these numbers tell us about the amount of photosynthesis that occurred in each of the two light conditions. In other words, was the hypothesis the class tested supported or not?
• Continue with a class discussion to analyse the data. How do you know that the bubbles you saw rise to the surface were bubbles of oxygen? Students may answer that they know photosynthesis produces oxygen, so the bubbles must have been oxygen. However, without a way to determine the chemical composition of the bubbles, it is only an assumption that the bubbles contain oxygen. They might just as well have been bubbles of nitrogen or carbon dioxide, or some other gas from some other process that was occurring in the plants instead of photosynthesis. Nevertheless, since the plants were exposed to light, the bubbles were most likely made of oxygen. Point out that it is important for researchers to make sure they recognize the difference between what they know about an experiment and what they assume about it.

mean: The sum of all the values in a set of data, divided by the number of values in the data set; also known as the average. For example, in a set of five temperature measurements consisting of 22 ºC, 25 ºC, 18 ºC, 22 ºC and 19 ºC, the mean temperature is 106 ºC divided by 5, or 21.2 ºC.

median: Tthe middle value in a set of data, obtained by organizing the data values in an ordered list from smallest to largest, and then finding the value that is at the half-way point in the list. For example, in a set of five temperature measurements consisting of 22º C, 25º C, 18º C, 22 º C, and 19º C, the ordered list of temperatures would be 18º C, 19º C, 22º C, 22º C, and 25º C. The middle value is the third value, 22º C. If the data set consists of an even number of values, the median is determined by averaging the two middle values. For example, in a set of six temperature measurements consisting of 20 ºC, 22 ºC, 25 ºC, 18 ºC, 24 ºC and 19 ºC, the middle values are 20 ºC and 22 ºC. Thus, the median value is the average of 20 ºC and 22 ºC, which is 21 ºC.

mode : The value in a set of data that occurs most frequently. For example, in a set of five temperature measurements consisting of 22 ºC, 25 ºC, 18 ºC, 22 ºC and 19 ºC, the measurement of 22 ºC occurs most frequently, so it is the mode. It is possible to have two or more modes in a set of data, if two or more values occur with equal frequency.

Questions : Evaluate students' comprehension by asking them questions such as:

• What "things" are needed in order for photosynthesis to occur?
• What are the products of photosynthesis?
• Where in the plant does photosynthesis occur?
• Why do plants need water in order to survive?

Graph Analylsis: Provide a graph of data from an experiment similar to the one students just performed, and ask them to draw conclusions from it. For example, the data could represent the heights of corn plants, half of which were grown in the shade of a forest and half of which were grown in an open field.

• What do you think would happen if you left some plants in a completely dark closet for two or three weeks? Why do you think that?
• Why is it important for crop plants to receive enough rainfall?
• The Earth's atmosphere did not always contain as much oxygen as it does now. In fact, at one time it probably contained no oxygen at all. How do you think the oxygen in the Earth's atmosphere got there? Why do you think that?

The light that comes from the sun consists of light waves of many different wavelengths. In the visible spectrum of light, these range from red with the longest wavelength, to violet with the shortest wavelength. Chlorophyll does not respond equally to all wavelengths, or colors of light. Have students use the same experimental setup to determine what color or colors of light result in the most photosynthetic activity. The only modification they need to make is to loosely cover the beaker with colored plastic wrap or cellophane during the five minutes of bubble counting. Since blue wavelengths are the best for most plants, be sure that this is one of the colors available. If possible, have red and one other color available as well.

Through a teacher-led discussion, students realize that the food energy plants obtain comes from sunlight via the plant process of photosynthesis. By counting the number of bubbles that rise to the surface in a five-minute period, students can compare the photosynthetic activity of Elodea in the pre...

Students learn about photosynthesis and cellular respiration at the atomic level and study the basic principles of electromicrobiology—a new field of research that may enable engineers to harness energy at the molecular level.

Contributors

Supporting program, acknowledgements.

This content was developed by the MUSIC (Math Understanding through Science Integrated with Curriculum) Program in the Pratt School of Engineering at Duke University under National Science Foundation GK-12 grant no. DGE 0338262. However, these contents do not necessarily represent the policies of the NSF, and you should not assume endorsement by the federal government.

Last modified: July 12, 2023

Using Elodea in the Lab

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This North American pondweed is widely recommended for demonstrating oxygen formation during photosynthesis.  Elodea  is cheap and easy to grow, but  other pondweeds  can also be used.

• Photosynthesis –  Resources: Demonstrating oxygen formation during photosynthesis can be a tricky process. One common way is to  gather bubbles of gas given off by an aquatic plant .

Teaching Topics

• Photosynthesis and respiration

 Description

Canadian Pondweed,  Elodea canadensis

A native aquatic plant of temperate areas of North America. A recorded pest on rivers and canals in the UK, particularly in the 19th century. Its distribution was aided by botanic gardens both in Britain and mainland Europe, and is now found widespread in all areas south of the Arctic, including Australia and New Zealand.

Growing and sourcing

Obtaining : Easily available from aquarium shops or pet shops that have aquarium section, and specialist science suppliers such as Blades Biological. A care sheet can be downloaded from them too. There are also species of  Elodea  available, some of which have been reclassified as  Egeria.

Care : Blades Biological provide care information for  Elodea .  They also supply both plants.  Elodea  can be kept living in an aquarium long term if anchored with gravel it should then begin to grow. Short term it can be kept in a shallow tray covered with water. You can find more information about caring for pondweed in the video  here .

• Photosynthesis
• Plants in the Science Curriculum

Related content

Teaching resources.

• A Leaf in Time - A Popular Introduction to Photosynthesis
• Photosynthesis - A Survival Guide for Teachers
• Measuring the rate of photosynthesis
• Investigating Photosynthesis with the SAPS / NCBE Photosynthesis Kit

• Compound & Stereo
• How to Use a Microscope
• Set Up Your Own Activities
• Low Power Stereomicroscopes
• Elodea leaf
• Human cheek
• Yeast bubbles
• Yoghurt bacteria
• Images & Videos
• Resources & Links
• M4S Competition

Elodea (pondweed) cells

• Glass microscope slides
• Plastic cover slips
• Paper towels or tissues
• Salt solution (6 g salt dissolved in 100 ml of water or approx. 2 teaspoons salt in a cup of water)

See information on suppliers here .

• Pick off an entire healthy looking Elodea leaf, with fingers or small scissors and place it on the microscope slide.
• Add a drop of water (hypotonic solution) and a coverslip and observe the chloroplasts (green structures) and the cell walls.
• Add a drop of salt solution (hypertonic solution) to the side of the coverslip and observe the cell shrinking (optional).

When the salt solution is added, the salt ions outside the cell membrane cause the water molecules to leave the cell through the cell membrane causing it to shrink into a blob in the centre of the cell wall. The movement of water molecules is called osmosis .

Elodea is a genus of submerged aquatic plants used in aquariums. You should be able to buy this from pet shops. If not, you can purchase it from a school supplier . Elodea canadensis is a species that works well for this activity, because it has thin, straight leaves. However, most other species are satisfactory.

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Teacher Guide: Elodea Photosynthesis Light

• Elodea sp. (or Egeria sp. often sold under the common name “anacharis”) in a container of fresh water
• Six 50 mL Erlenmeyer flasks
• Six one-hole #2 rubber stoppers
• Six 1 mL graduated glass serological pipettes
• Red and green light filters
• Aluminum foil
• Thermometer
• Fig. 2.40 and 2.41
• Blue light filter (optional)
• Petroleum jelly (optional)

Fig. 2.40. This diagram of the electromagnetic spectrum emphasizes the small portion of the spectrum that is visible to human eyes. Wavelengths are measured in meters (m) along the grey bar and in nanometers (nm) along the colored bar showing visible light.

Image courtesy of Philip Ronan, Wikimedia Commons

Fig. 2.41. Relative absorbance of the visible wavelengths in sunlight by the pigments chlorophyll a and chlorophyll b

Image courtesy of Daniele Pugliesi and M0tty, Wikimedia Commons

Baking Soda Solution

Baking soda serves as a source of carbon dioxide. The Elodea will be placed in this solution to speed up the photosynthesis reaction .

• Baking soda (sodium bicarbonate, NaHCO 3 )
• Fresh water
• Elodea is a genus of freshwater aquatic plants sold in pet stores for aquariums. Observe and describe the Elodea specimen.  
• Use Fig. 2.40 to determine the range of wavelengths that corresponds to white light and to each color: red, blue, and green. Fill in the first column of Table 2.8.  
• What color(s) does chlorophyll a absorb most? Least?
• What color(s) does chlorophyll b absorb most? Least?  
• Using what you know about the electromagnetic spectrum and chlorophyll, predict the flask that will have the plant with the highest gas production and write “1” in the “Prediction” column of Table 2.8. The more light energy the plant absorbs, the more gas it should produce.
• Write number “2” in the “Prediction” column of Table 2.8 for the flask that you predict will have the second highest gas production.
• Continue numbering in the “Prediction” column of Table 2.8 through “6,” with 6 being the flask that you predict will have the least gas production.  

Photosynthesis should be fastest with ____________ wavelength(s) of light and slowest with ____________ wavelength(s) of light because ________________.  

• Weigh 25 g of baking soda.
• Add baking soda to 1 L of fresh water.
• Stir until the baking soda is completely dissolved.  
• Cut out rectangles of green and red filter paper slightly bigger than the flasks.
• Wrap filters around the flasks, secure with tape. Trim excess filter.
• Wrap one flask with foil.  
• Remove a few branches of Elodea from the holding container. Visually inspect the Elodea, and remove any part of the plant that looks unhealthy or has different leaf morphology (shape) than the rest of the plant. Blot Elodea dry with towels.  
• Using a balance, weigh 2.5 g of Elodea . The Elodea should be as close as possible to 2.5 g, within 0.1 g (i.e., between 2.4 and 2.6 g). Write the weight in Table 2.8.
• Insert Elodea into the flask. If necessary, you can break the Elodea into smaller pieces and use a skewer to distribute the Elodea evenly in the flask.  
• Working over towels, slowly fill each flask all the way to the top with baking soda (sodium bicarbonate) solution.
• Firmly press a stopper into each flask. Sodium bicarbonate solution will spill out of the flask as you insert the stopper. Applying petroleum jelly to the outer surfaces of the rubber stoppers may help to form an airtight seal.
• Insert the pipette into the stopper hole by holding the pipette with a dry towel. Gently twist the tapered tip of the pipette into the stopper until water rises in the pipette and the pipette is firmly in place. The water level should reach between the 0.8 and 0.7 mL lines. DO NOT FORCE the pipette into the stopper as the pipette can break. If you are having difficulty getting the pipette into the stopper, ask your teacher for assistance.
• Dry the outside of each flask with a towel.  
• Set the flasks in the sun. Make sure each flask is exposed to a similar amount of sunlight.  
• Record the starting volume of the liquid in each pipette and the time in Table 2.8. Read the bottom of the meniscus of the water in the pipette. Note that the numbers on the pipette are smallest at the top and largest at the bottom.  
• record the weather conditions, especially noting the amount of sunlight, and
• observe what is happening in the pipettes and the flasks without disturbing the light filters or foil.  
• At the end of the experiment, record the ending volume (water level) in each pipette and the time in Table 2.8.  
• Subtract the start volume from the final volume to get the total amount of gas produced in each flask. Record the amount in Table 2.8.  
• Rank the gas production in each flask, by writing “1” in the “Observation” column for the flask with the plant the produced the most gas and numbering through “6” for the flask that produced the least gas.
• OPTIONAL: repeat procedure steps 7 to 16 using a blue light filter.  
• Holding the pipette with a dry towel, gently twist the pipette while pulling up to remove it from the stopper. Stand the pipette upright in a container and let it drip dry.
• Remove the stopper from the flask.
• Dispose of the plants and sodium bicarbonate solution as directed by your teacher. Clean each flask.
• Wipe the light filters with fresh water to remove traces of sodium bicarbonate solution.  
• Compare your results to those of other groups using a class data chart.
• What caused the water in the pipettes to rise?  
• What gas is being produced in the flasks by Elodea ? What process is producing this gas?  
• Compare your predictions and observations. Was your hypothesis supported? Why or why not? Give you answer in terms of absorption of wavelengths by chlorophyll pigments.  
• How did your results compare to those of the rest of the class? Hypothesize possible reasons for any unexpected results.  
• What gas is likely being produced?
• What process is likely producing this gas?  
• What was the purpose of the clear flask with no plant? In other words, is the production of gas the only thing that may affect the starting and ending water volume in the flasks?  
• How can you use the results of your control flasks to more accurately calculate gas production in the other flasks?  
• How can this experiment be improved to more accurately measure the photosynthesis rate in Elodea ? What sources of error can be further controlled?  
• Why are plants green? Answer in terms of the wavelengths of light they absorb. (Hint: What colors do chlorophyll a and chlorophyll b absorb and reflect?)  
• Based on Fig. 2.40, if green glass is a green-yellow color, what range of wavelengths might it be reflecting?  
• Based on Fig. 2.40 and Fig. 2.41, what color(s) would the pigments chlorophyll a and chlorophyll b appear to the human eye?

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Science project, elodea photosynthesis lab.

Grade Level: 7th - 10th; Type: Life Science

Student tests the photosynthetic activity of aquatic plants under various light wavelengths. 

Research Questions:

• How do plants perform photosynthesis?
• What gases are used by plants during photosynthesis?
• What gases are produced by plants during photosynthesis?
• What is the role of chlorophyll in photosynthesis?
• Which wavelengths of light are used by chlorophyll in plants during photosynthesis?

Plants perform photosynthesis to create sugars and other organic compounds that they need to survive. Chlorophyll is a pigment in the chloroplast of the plant cell that absorbs energy from light and uses it to create compounds needed in photosynthesis. Visible light is made up of different colors of different wavelengths and energy. Certain wavelengths of light are used by plants more than others. Plants use carbon dioxide and produce oxygen gas during photosynthesis. They produce carbon dioxide during cellular respiration. In this experiment, the student will place aquatic plants under different colors of light in a solution of bromothymol blue.   Bromothymol blue is an acid-base indicator that turns yellow in acidic solutions and blue in alkaline solutions. Carbon dioxide bubbled into water forms carbonic acid. Thus, the more photosynthetic activity, the less acidic and more blue the bromothymol blue solution becomes as the plant uses carbon dioxide during photosynthesis. The less photosynthetic activity, the more acidic and yellow the bromothymol blue solution becomes as the plant produces carbon dioxide during cellular respiration.

• Elodea, an aquatic plant available at many pet or gardening stores
• Bromothymol blue solution (acid-base indicator available for purchase online) 0.04% (Aqueous), 1 L Bottle (1-2 bottles)
• Transparent plastic films of different colors (heat resistant)
• Masking tape
• Clamp lights (one for each color and one additional for the control)
• Test tubes or small glasses (one for each color and two additional as controls)
• Plastic wrap
• Aluminum foil

Experimental Procedure:

• Cover each clamp lamp with a colored plastic film. Secure the film with tape. Be sure that the film is heat resistant to avoid fire hazard. Leave one lamp without a colored film as a control. Set up the lamps at least several feet apart and away from windows. Write a hypothesis to explain which color light will be best for photosynthesis and why.
• Pour equal amounts of bromothymol blue into the test tubes or glasses (about 2/3 of the test tube or ½ cup in a small glass). If you do not have enough solution, you may dilute it with a little water. Just be sure to use distilled water and dilute the entire supply only slightly. Write the name of the colored film to be tested on each cup (you can also use masking tape to label cups). 
• Cut equal-sized pieces of elodea for each test tube or glass (about 3 inches in length). Make sure that each piece looks healthy and has plenty of green leaves. Place a piece of elodea in each test tube or glass. Record the initial color of the bromothymol blue solution. Cover each test tube or glass with plastic wrap. Be sure to completely seal the vessel to keep gas from entering or leaving. For one of the control plants, cover the test tube or glass completely with aluminum foil to block out any light. 
• Place one uncovered plant and the covered plant under the light without any colored film. Place the remaining plants each under a colored lamp. The plants should all be 12 inches (30 cm) away from their lamp. Allow the plants to remain under the lamps until the uncovered control has turned dark blue (about 24-48 hours). Record the final color of the bromothymol blue solution for each color of light, as well as both controls. Create a bar graph to illustrate your results. Compare your results to your hypothesis.

Terms/Concepts:  Photosynthesis; Cellular respiration; Visible light spectrum; Chlorophyll; Carbon dioxide; Bromothymol blue

References:

• http://wardsci.com/product.asp_Q_pn_E_IG0015113
• www.mysciencebox.org/files/bromthymol_blue.doc
• http://www.amphi.com/teachers/crobson/files/BAAAB5B835C54A3BB7C470B96B082B19.pdf
• http://www.msnucleus.org/membership/html/k-6/lc/plants/5/lcp5_3a.html

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Laboratory Manual For SCI103 Biology I at Roxbury Community College

9 photosynthesis.

In this lab, we will study the effect of light intensity and quality (wave length - color) on photosynthesis . As a measure of the rate of photosynthesis, we will monitor the rate of oxygen production. When plants that spend their life submerged in water release oxygen it forms bubbles, which we can count over a period of time to determine photosynthesis rate.

Photosynthesis is a process used by plants and other organisms to convert light energy into chemical energy that can later be released to fuel the organisms’ activities (energy transformation). This chemical energy is stored in carbohydrate molecules, such as sugars, which are synthesized from carbon dioxide and water - hence the name photosynthesis, from the Greek phōs, “light”, and synthesis, “putting together”. In most cases, oxygen is also released as a waste product. Most plants, most algae, and cyanobacteria perform photosynthesis; such organisms are called photoautotrophs. Photosynthesis is largely responsible for producing and maintaining the oxygen content of the Earth’s atmosphere, and supplies all of the organic compounds and most of the energy necessary for life on Earth.

Although photosynthesis is performed differently by different species, the process always begins when energy from light is absorbed by proteins called reaction centers that contain green chlorophyll pigments. In plants, these proteins are held inside organelles called chloroplasts, which are most abundant in leaf cells, while in bacteria they are embedded in the plasma membrane. In these light-dependent reactions, some energy is used to strip electrons from suitable substances, such as water, producing oxygen gas. The hydrogen freed by the splitting of water is used in the creation of two further compounds that act as an immediate energy storage means: reduced nicotinamide adenine dinucleotide phosphate (NADPH) and adenosine triphosphate (ATP), the “energy currency” of cells.

In plants, algae and cyanobacteria, long-term energy storage in the form of sugars is produced by a subsequent sequence of light-independent reactions called the Calvin cycle; some bacteria use different mechanisms, such as the reverse Krebs cycle, to achieve the same end. In the Calvin cycle, atmospheric carbon dioxide is incorporated into already existing organic carbon compounds, such as ribulose bisphosphate (RuBP). Using the ATP and NADPH produced by the light-dependent reactions, the resulting compounds are then reduced and removed to form further carbohydrates, such as glucose.

The first photosynthetic organisms probably evolved early in the evolutionary history of life and most likely used reducing agents such as hydrogen or hydrogen sulfide, rather than water, as sources of electrons. Cyanobacteria appeared later; the excess oxygen they produced contributed directly to the oxygenation of the Earth, which rendered the evolution of complex life possible. Today, the average rate of energy capture by photosynthesis globally is approximately 130 terawatts which is about three times the current power consumption of human civilization. Photosynthetic organisms also convert around 100-115 thousand million metric tons of carbon into biomass per year.

The main source of light on Earth is the Sun. Sunlight provides the energy that green plants use to create sugars mostly in the form of starches, which release energy into the living things that digest them. This process of photosynthesis provides virtually all the energy used by living things. The primary properties of visible light are intensity, propagation direction, frequency or wavelength spectrum, and polarization, while its speed in a vacuum, 299,792,458 meters per second, is one of the fundamental constants of nature. Visible light, as with all types of electromagnetic radiation (EMR), is experimentally found to always move at this speed in a vacuum.

9.1 Intensity of light

Light is electromagnetic radiation within a certain portion of the electromagnetic spectrum (Figure 9.1 ). The word usually refers to visible light, which is visible to the human eye and is responsible for the sense of sight. Visible light is usually defined as having wavelengths in the range of 400-700 nanometres (nm), or 400 × 10 -9 to 700 × 10 -9 m, between the infrared (with longer wavelengths) and the ultraviolet (with shorter wavelengths). This wavelength means a frequency range of roughly 430-750 terahertz (THz).

Figure 9.1: Spectrum of light. V, violet; B, blue; G, green Y, yellow; O, orange; R, red

In this experiment (Figure 9.2 ), we will study the effect of light intensity on the photosynthetic activity of Elodea canadensis . We will vary the light intensity by changing the distance between the light source and the plant. We will count the emerging oxygen bubbles as an indicator of the photosynthetic activity of the plant.

Figure 9.2: Setup for photosynthesis experiment.

9.1.1 Experimental procedures

Before you begin with the actual experiment, write down in your own words the hypothesis for this experiment:

• Obtain a cylindrical test tube.
• Fill test tube with 0.3% sodium bicarbonate.
• Select a fresh, crisp sprig of Elodea about 15 cm in length.
• While the plant is still submerged, cut 2-3 mm from its base.
• Place the sprig upside down into the test tube filled with sodium bicarbonate. The sodium bicarbonate will absorb anu toxic materials that are released by the plant during photosynthesis.
• Keeping the plant submerged, position a light source 10 cm away and adjust so the light shines directly on the plant.
• Place the test tube in a beaker of water as shown in Fig. 9.2 to prevent overheating the plant. 1. 1. Allow the system to stand 7-10 minutes, or until bubbles begin to appear regularly.
• Count the bubbles produced each minute for a 5-minute period and average them. Record your findings in the table.
• Move the light back 20 cm from the plant, wait 5 minutes, and repeat counting. Record your findings in Table 9.1 .
• Move the light back 40 cm from the plant and repeat counting the bubbles.
• When you have finished recording your data, calculate the average number of bubbles for each 5 minute period and enter the result into the table.

Do the data support or contradict your hypothesis?

Figure 9.3: Appearance of bubbles indicates active photosynthesis.

9.2 Color of light

In this experiment, we will study the effect of the color of light on the photosynthetic activity of Elodea canadensis . We will use filter to expose the plant to light of only a limited range of wavelengths. We will again count the emerging oxygen bubbles as an indicator of the photosynthetic activity of the plant.

9.2.1 Experimental procedures

• Empty the test tube that you used in the previous experiment.
• Fill the test tube with fress 0.3% sodium bicarbonate.
• Place the Elodea sprig into the test tube and submerge it completely in the bicarbonate.
• Place the red colored filter between the test tube and the heat shield beaker and allow it to sit for 5 minutes.
• Count bubbles for 5 minutes as in the previous experiment. Record your findings in Table 9.2 .
• Remove the color filter and expose the plant to white light. Count bubbles again for 5 minutes in 1 minute intervals. Record your findings in Table 9.2 .
• Place the green colored filter between the test tube and the heat shield beaker and allow it to sit for 5 minutes.
• Count bubbles for 5 minutes. Record your findings in Table 9.2 . Table: (#tab:color) Color of light.

9.3 Determination of the light absorption spectrum of dye solutions

In this experiment, we will use a spectrophotometer to measure the differential absorption of light of different wavelength by water stained with food dyes.

Figure 9.4: Spectrophotometer and cuvettes with dye solutions.

9.3.1 Experimental procedures

• Take six cuvettes.
• Fill one cuvette with water.
• Fill each of the remaining five cuvettes with one of the color solutions listed in Table 9.3 .
• Insert the cuvette with water into the slot marked “B”.
• Insert the other cuvettes into the slots marked 1 to 5 and write down which color is in which slot.
• Following the instructions posted on the spectrophotometer, program the machine to take absorption measurements at wavelengths between 380-740 nm in 20 nm steps.
• Once the measurements are completed, write down the absorption number for each dye and wavelength.
• Use a spreadsheet program to graph your results.
• Compare your curves with the data shown in Figure 9.6 .

Figure 9.5: Cuvettes placed in the spectrophotometer.

Figure 9.6: Normalized absorption of red, green and blue dye solutions. Compare these data with your own results.

9.4 Chromatography

Chromatography is a laboratory technique for the separation of a mixture. The mixture is dissolved in a fluid called the mobile phase, which carries it through a structure holding another material called the stationary phase. The various constituents of the mixture travel at different speeds, causing them to separate. The separation is based on differential partitioning between the mobile and stationary phases. Subtle differences in a compound’s partition coefficient result in differential retention on the stationary phase and thus affect the separation. Chromatography may be preparative or analytical. The purpose of preparative chromatography is to separate the components of a mixture for later use and is thus a form of purification. Analytical chromatography is done normally with smaller amounts of material and is for establishing the presence or measuring the relative proportions of analytes in a mixture.

In this experiment, we separate a mixture of food dyes (a dark brown liquid). The mobile phase (separation buffer) is 1% NaCl in water, the stationary phase is chromatography paper.

9.4.1 Experimental procedures

• Obtain a small beaker.
• Add NaCl running buffer to the beaker until it reaches a height of about 5 mm.
• Obtain a strip of chromatography paper and put it down on the bench.
• Obtain the bottle containing the dark green food dye mixture.
• Obtain a glass capillary and insert the tip of the capillary into the food dye mixture liquid. A little bit of dye will ascend into the capillary.
• Remove the capillary and apply.
• Touch the left side of the chromatography paper about 1 cm above its lower end with the tip of the capillary. A little bit of green liquid will spread out on the paper. Lift the capillary and touch the paper again just to the right of the dye you just applied. Repeat this until you have a horizontal line of dye from the left to the right side of the paper.
• Place the chromatography paper into the beaker as shown below.
• Observe how the running buffer moves up the paper and separates the dye mixture into three components (red, yellow and blue.

Figure 9.7: Result of the Chromatography experiment.

9.5 Review Questions

• What is light?
• In your own words, describe the endproducts of photosynthesis.
• In your own words, describe what happens in photosynthesis.
• What is chlorophyll and what does it do?
• Where inside of plant cells does photosynthesis happen?
• What is chromatography and what is it used for?

The Biology Corner

Biology Teaching Resources

Observe Photosynthesis with this Easy Experiment

Photosynthesis is a complicated topic that requires students to develop mental models of the phenomenon. Teachers may struggle to find hands-on activities that can be completed in a short class period.

There are several ways this can be accomplished. Students can observe the products created by photosynthesis, oxygen. In this lab, students cut small disks from spinach leaves and record the time it takes to float.

You can also indirectly measure photosynthesis by the amount of carbon dioxide absorbed during the process. I have another lab that can be a demonstration for the whole class where students can see the effects of photosynthesis. In this demonstration, an indicator ( phenol red) is used to measure the amount of carbon taken up during the process.

Class Demonstration

The demonstration is simple. You take leaves from a plant and place it in the indicator solution. The plants exposed to light will consume the carbon dioxide which decreases the acidity of the solution. The yellow solution will turn red!

I have used elodea leaves in the past, but find that these specimens have become hard to acquire. Many states consider this water weed an invasive species and limit shipping. Luckily, leaves from kale will also work. Kale is a fairly inexpensive vegetable you can get at the supermarket.

The demonstration is simple. Phenol red starts out as a red color but will change to yellow if you blow into it with a straw. The carbon dioxide in your breath will change the solution to a yellow color.

Next, place leaves from kale into test tubes with the yellow solution. Place one tube in the light and another in the dark (using aluminum foil).

A full spectrum grow light will have the best results. Leave one test tube empty as a control. In about 20-30 minutes, the test tube in the light will change to red, indicating that the carbon dioxide has been consumed.

You can use this demonstration as an introduction to photosynthesis, or a plant unit. Follow up with a discussion that asks students the following questions:

• Why does phenol red change color when I add carbon dioxide?
• Why does it turn back to red in the tube with a plant exposed to light?
• Why did the plant in the dark not change color?

If you want a more student-directed activity, students can do the lab for themselves. This handout outlines the procedure and includes discussion questions for students to answer in groups.

As an extension activity, ask students what will happen to the tubes if left to sit overnight. Generally, the color will change to yellow as plants respire and release carbon dioxide. (Note: I have had mixed results with this, but it’s a good way to prompt discussion about the relationship between respiration and photosynthesis.)

Shannan Muskopf

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Care Guide: Elodea

Living care information.

Elodea , Egeria Commonly known as waterweed

Quick Start Information

Brazilian Elodea is frequently referred to in the classroom and in the aquarium trade as Elodea, but it is properly classified as Egeria densa . In this guide Elodea refers to both Egeria densa and Elodea canadensis .

We ship Elodea ( Egeria densa ) in packs of 12, 25, or 50 plants in plastic bags without water.

The plant is also available as Elodea tips in a jar of water.

Elodea can vary in color from dark brown to bright green depending on the time of year and conditions at the collection site.

We collect our Elodea. Due to this, there may be other organisms present in your shipment.

About the Organism

• There are Elodea species native to both North and South America.
• Elodea plants live underwater except when small white flowers bloom at the water’s surface to produce seeds.
• Elodea’s main reproductive strategy occurs when stalks detach from the parent plant, float away, root and produce new plants.
• Elodea beds provide an important nursery habitat for aquatic invertebrates and young fish.
• Brazilian Elodea ( Elodea densa ) is an invasive species in many parts of the world.
• Domain: Eukarya
• Kingdom: Plantae
• Phylum: Magnoliophyta
• Class: Liliopsida
• Order: Hydrocharitales
• Family: Hydrocharitaceae
• Genus: Egeria or Elodea
• Species: densa, canadensis, najas

Preparation

Elodea can be purchased year-round. Elodea is a freshwater organism and should be kept in dechlorinated tap water. Water from the tap in most municipalities contains chlorine and chloramines which are not removed by off-gassing. Water should be treated with a dechlorinating agent .

Since our Elodea is collected, a range of color can be expected, anywhere from emerald green to brownish, depending on growing conditions at the collection site. As long as the foliage is firm and not shedding excessively it is healthy and ready for use.

If you intend to use Elodea in a laboratory investigation, you can use the plants immediately after rinsing them. For optimum results of your investigations, we recommend transferring them to water and putting them under lights for 2 days before using them. This allows the plants time to recover from any shipping shock.

Shallow containers with a broad surface area are best for holding plants until they are used for the lab. Use either tap water treated to remove chlorine and chloramines or spring water. We do not recommend distilled or deionized water, as these lack minerals needed by the plants. Clip the rubber bands from the stems and spread the plants into a single layer in the water. A fluorescent light bank or a 23- to 32-W compact fluorescent bulb in a desk lamp will provide the intense light needed by the plants. If you must keep the plants for several days before lab use, replace about 1/4 of the water every third day. This is to prevent mineral depletion.

If you plan on using your Elodea as an aquarium specimen, provide the habitat with 10 to 12 hours of light per day at 3 to 5 watts per gallon. Keep the habitat at temperatures ranging from 50 to 77° F for Egeria densa , and 45-65°F for Elodea canadensis . Your container should be large enough to submerge the entirety of the Elodea in treated water. Elodea can grow if it is free floating, but it does grow more vigorously if it is rooted in a substrate.

Depending on the make-up of your aquarium community, it may be necessary to add an aquatic plant fertilizer to replenish depleted mineral content. Iron is of primary concern as Elodea depletes this mineral rapidly. Leaf chlorosis is the most common sign of iron deficiency. Leaves yellowing at the edges only may be showing signs of potassium deficiency.

Aquatic plant fertilizers are available from many aquarium supply or garden shops. Follow the manufacturer’s instructions for adding the fertilizer to your aquarium.

If the aquarium also contains animal inhabitants, phosphate and nitrate levels should also be monitored to ensure the continued health of the aquarium community.

Maintaining and culturing

Specimens of Elodea used for a lab can be propagated in an aquarium as described above.See the Housing (Aquarium) section.

Egeria densa is considered invasive in many parts of the world. Carolina provides living organisms for educational purposes only. As a general policy, we do not advocate the release of organisms into the environment. In some states, it is illegal to release organisms, even indigenous species, without a permit. The intent of these laws is to protect native wildlife and the environment.

Invasive aquatic vegetation is a conservation issue in many areas, therefore, any plants that are no longer needed should be placed in a sealed container and frozen for 72 hours. Dispose of materials according to local and state guidelines.

Please contact your local (state) Department of Agriculture for any specific restrictions on release and disposal of organisms.

No biosafety information applies for this organism.

How can I use Elodea to study photosynthesis?

During photosynthesis, the plant absorbs carbon dioxide from the water and releases oxygen. Since oxygen is much less soluble in water than is carbon dioxide, the water quickly saturates with oxygen and bubbles form. The number of bubbles released per minute measures the rate of photosynthesis. Alternately, use a simple volumeter to measure the increase in volume of gas caused by the release of oxygen.

What is the difference between Elodea and Elodea Tips ?

Elodea consists of Elodea sprigs that are about 11 cm (4 ¼") in length. We recommend these for aquarium plants and for studies of photosynthesis, although they can be used also for cell studies. Elodea Tips are the terminal 3 to 4-cm ends cut from sprigs. We recommend these as best for cell studies.

Why can’t you ship me Elodea ?

Some states restrict us from shipping Elodea densa because they consider it an invasive plant. We usually substitute Elodea canadensis , a native non-invasive plant, for orders going to restricted states; however, there are times when Elodea canadensis is not available and we must substitute another plant. Never release a non-native plant or animal into your local environment. To discard an aquatic plant, place it in a plastic bag and freeze it for at least 72 hours before discarding the unopened bag in the trash.

Can I substitute other plants for Elodea densa?

For photosynthesis studies, almost any aquatic plant will work. These include Ludwigia , Cabomba , Myriophyllum , Ceratophyllum , and Sagittaria . For easy viewing of plant cells with chloroplasts, consider Fern Prothallia .

We want you to have a good experience. Orders and replacements: 800.334.5551, then select Customer Service. Technical support and questions: [email protected]

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