A Simple Method to Explore Enzyme Activity

A Simple Method to Explore Enzyme Activity

by Brad Williamson

Enzymes are essential in nearly all life processes. By catalyzing (speeding up a chemical reaction) various chemical reactions in the cell enzymes make energy and nutrients available to living systems. Each enzyme only catalyzes one kind of chemical reaction so there is a lot of enzymes in a typical cell. As the human genome project uncovers the genes in the human population it is important to realize that many of the genes code for enzymes. A basic understanding of enzyme properties then is an essential, first step, to understanding living systems.

Enzyme studies are easily studied by students. For this investigation we will work with an enzyme that begins the breakdown of starch into sugar. It’s called amylase (enzyme) since it breaks down starch (the substrate) which is known as amylose. Scientists use the term substrate to describe the chemical that an enzyme catalyzes. Starch is an energy-storing compound present in many plant foods such as potatoes, corn and bread. Amylase is a component of saliva where it starts the digestion of starch as you chew your food. It’s also a valuable commercial enzyme. The starch in grains such as sorghum are digested with amylase in large reaction vessels to break the starch down to sugar which is then used to produce a number of products. Quite a bit of research that is done by various agricultural product companies attempts to find more efficient forms of enzymes such as amylase. Any organism that might consume starch (including plants) probably relies on some form of this enzyme. And each organism might have just a slightly different form of the enzyme that might be more efficient than others. You might find such an organism and find a valuable enzyme product.

Most of the time biochemists (scientists that study enzymes) work with enzymes in a liquid environment. Such techniques require careful control of a number of variables. A simpler method to study enzymes involves a gel-like substance known as agar. The substrate (starch in this case) is dissolved in this gel and various suspected enzyme-containing substances are added to small holes (wells) in the gel. The suspected enzyme diffuses out through the gel. If it can actively digest starch it will create a starchless area around the well. Iodine stain can be used to cause starch to turn a dark purple. Clear zones that are not purple are areas that the enzyme has digested the starch to sugar. This technique makes it simple to test many samples for activity or to determine the amount of activity a specific enzyme might have. In addition, it is easily modified to test various variables that might affect enzyme activity.

You’ll find the entire lab in pdf format for downloading here: A Simple Method to Explore Enzyme Activity

Ant Lions and Biology

Ant Lions and Biology

by Brad Williamson

Science is driven by questions. For students to really experience the process of science they should work on answering questions that arise from their own natural curiosity. Ant lions (Neuroptera) and their intriguing behaviors naturally generate student questions. Since most students are not familiar with “doodle bugs”, ant lions make an effective way to open the school year. If a few students have prior experience with ant lions this only adds to the effectiveness of the activity.

You’ll find the entire lab in pdf format for download here: Ant Lions and Biology




by Lisa Volland

A great way to end a semester of studying the different kingdoms is to have a scavenger hunt! This hands-on lab practical exam will allow students to take their knowledge of the various types of plants, fungi, and animals into the field. In my regular and honors biology, mostly made up of freshmen and sophomores, we study the diversity of organisms by doing lab or field observations of living specimens during the last part of the second semester. If it is sometimes difficult to get outside in the spring because of location, transportation, weather, or otherwise–bring the outdoors inside with samples of wildflowers, mosses, leaves, earthworms and other small invertebrates and vertebrates. Practice conservation by collecting only what you intend to use. Return living specimens to their habitat whenever possible.

Below is a modified scavenger hunt appropriate for eastern Kansas. This originated with John Wachholz, Salina Central, and Ernie Brown, WaKeeney, fellow KABT members. Some of the following items we have looked at in class, others not, but they must decide on where to go look for the specimen and how to collect it. I have added to this list and taken other items away as some choices are either too easy or too difficult to collect.

I have my students take this list two weeks before the end of school. They may begin checking in the next morning or after school. I stop answering questions about the project after I make sure that all the directions are understood. As the students check in their items, I initial the circle, then throw away the item, or if it is an animal, the student has to decide where to put it in my room and how to keep it alive. We feed the live insects and fruit to the classroom animals
We have a seminar period built into our block schedule twice a week, so I allow those that missed checking in that morning to check in during this period. Most students are able to finish this lab practical on time, and with excellent scores. The total points assigned to this final is 15% of the semester grade points.

The lab (including the student scavenger hunt form) in pdf format for download: BIOLOGY LAB PRACTICAL – THE SCAVENGER HUNT

Quiz Card Dissections

Quiz Card Dissections

by Ernie Brown

Here’s a pdf version for download: Quiz Card Dissections

Do you hate to think about doing classroom dissections in biology because the students tend to get unruly? Does your room resemble a zoo more than a place where learning is taking place? Do you feel that your students really appreciate the inherent value of the life of the animal that they’re dissecting. Do they appreciate the fact that the animal they are dissecting died so that they might learn more about how it lived? Respect for life and learning more about how the organisms lived are important objectives for any dissection. Too often specimens are viewed simply as “pieces of meat” to glibly chop up and discard at the end of the class period. I have enjoyed great success with a “Quiz Card” approach to dissections for the last several years. I feel that the students learn the material better and appreciate the total functioning organism more completely when they have finished their dissection. The technique can be used with any textbook or lab handout that supplements your dissection activity.

While the students are reading the procedure for the dissection, make a list of all of the organs they are expected to locate and learn about on the chalkboard. Then, write the name of each organ on a separate 3 X 5 card and show the class your “deck of cards”. Have the students work in pairs, either determined by the teacher or by the students themselves. After the students have completed the dissection and are confident they know the location and functions of each of the listed organs on their own specimen, they sign up on the chalkboard indicating they are ready for their quiz over the material.

Beginning with the first pair of names on the list, take your 3 X 5 cards to their desk and “make a deal” with them for their quiz. I usually make the quiz worth 20 points total and have each student of the pair pull two cards (face down) from the deck. Each 5 point card identifies the organ that student must locate and discuss without help from his teammate. After the first student has completed his 10 point portion of the quiz, the second student then locates and discusses the two organs on the cards he has selected. Each member of the team receives the composite score from their individual quizzes.

You can vary this activity easily by making the quiz worth more or less than 20 points. You can let each pair of students make their own “deal” by picking any combination of cards/points that meet the total points for the quiz. Sometimes the students like to “go for broke” and pick one card for 10 points or pick five cards for two points each. When students realize that they are going to be required to locate and discuss specific organs in their specimen, they are much more diligent during the dissection. Procedures are read thoroughly and cuts are made carefully leaving organs in place as much as possible rather than being removed and piled on the dissecting tray.

It is the responsibility of each team to conduct the dissection and learn the organs by working together. Even if one member of the team doesn’t want to touch the specimen, he is still accountable for using a dissecting needle to point to the organs on his 3 X 5 cards. They can still be involved in the dissection by reading the procedure to the person actually conducting the dissection. By combining the individual quiz scores for a total team score, the students work together to teach each other much better since they each have an investment in the final quiz score.

Ernie Brown – Trego Community High School,Wakeeny, KS 67672

Clearing and Staining Small Vertebrates

Clearing and Staining Small Vertebrates

By Ernie Brown

Here’s a pdf copy to download: Clearing and Staining Small Vertebrates


When teaching the skeletal system in anatomy class or comparing the structure of homologous organs, it is useful to be able to make comparisons between skeletal features of different vertebrate organisms. The bones to be studied can be carefully dissected away from the muscle tissue and then reassembled, requiring a great deal of teacher or student preparation time. By use of the clearing method, however, the entire skeleton can be studied in relation to the muscles, circulatory system, and other organ systems. After a little experience with the procedure, it is a simple matter to introduce modifications which will improve the quality of the preparations. At times, two specimens, treated in a similar manner, will respond differently.
Fresh specimens may be preserved in 10% formalin solution or 75% alcohol for a period of three to five days. Generally, specimens fixed in formalin are more easily controlled and require less watching. Formalin fixed tissues often require a longer time for the clearing process and the specimen may not clear as completely as alcohol fixed specimens. A little experimentation will provide the best results for the type of specimens you wish to clear.

    Completely eviscerate the specimen if the skeletal system is to be demonstrated.

    Place the specimen in 2% KOH (potassium hydroxide) or 2% NaOH (sodium hydroxide). This step will decolorize the tissue and make it jelly-like in consistency. Use the same clearing solution for both alcohol and formalin fixed specimens. You can control the rate of tissue maceration by adjusting the temperature of the KOH or the room temperature. As the pigments are dissolved out of the tissue, the solution may become discolored, in which case it should be changed as often as necessary.

    Staining the skeleton begins after the flesh has become cleared and the bones are visible through the surrounding tissue. Alizarine red S is the dye recommended by most authors but others also have used indigo-carmine, Bordeaux red, alizarol black3G, and alizarine black SBB. Make a saturated stock solution of alizarine red S and add it to the 2% KOH solution to make it reddish in color. Cover the specimen with the dye solution and leave it to set.

    The specimen will usually stain completely within two days but additional time may be required for larger specimens. After staining, the specimen is placed in clear 2% KOH solution and destained until the muscle tissue is again clear and the stain remains only in the bones. If the bones were not stained completely, repeat the staining process and clear as before.

    When the specimen looks nearly transparent, place it in glycerine for storage. This will complete the clearing process. The change from 2% KOH to pure glycerine must be gradual for large specimens but is not important with smaller ones. A half and half mixture of pure glycerine and 1% KOH is recommended for those large specimens.


When clearing fish, the scales must be removed in order to see the skeleton. Scaling is best done after the fish has been stained since the scales will be red and easily visible. Some lizards must also be scaled. The scales may be easily removed by gentle scraping in a dish of water with a wire loop or other similar instrument. The hair of mammals and the feathers of birds may best be removed prior to staining.
• Davis, D. Dwight and V. R. Gore. Clearing and Staining Skeletons of Small Vertebrates. Chicago Nat. Hist. Mus. Technique Ser., 16p. 1947
• Evans, Howard E., Turtox News, Vol. 26, No.2, February 1948
• Mayorga, Horacio. A Rapid Method for Clearing and Staining Amphibian Skeletons Journal of The Ohio Herpetological Society, Volume 5, Number 1, May 1965