BI 101

Introduction to Biology
Laboratory Manual
BI 101 LAB 1
Measurement and the Metric System

Prepared by Dr. Emily Kasl and Dr. Eric Becraft, Dept. of Biology, UNA

OBJECTIVES
After completing these laboratory activities, you should understand / be able to:
• Recognize the standard (base) metric units and their abbreviations: meter, liter, gram, degrees
Celsius and metric prefixes (micro-, milli-, centi-, deci-, kilo-).
• Define density and calculate the density of seawater and tap water.
• Use metric units to measure length, volume, mass, and temperature and convert between
measurements (e.g. from meters to centimeters).
• Recognize and use metric rulers, graduated cylinders, beakers, a triple-beam balance, thermometer.
• Explain the advantages of the metric system of measurement.

Introduction
Devised in 1791 by the French Academy of Table 1. Standard units of the metric system.
Sciences, the metric system arose from the Standard (Base) Unit Abbreviation
need to standardize units of measurement
for research comparisons. In the 1960s, the Length Meter m
metric system became the basis for the Volume Liter L
International System of Units (SI units) that Mass Gram g
standardized units for length, volume, mass,
and temperature (Table 1). Temperature Celsius °C

Metric measurements are based on units of 10 (the prefix deci- means tenth). Metric units may be 10,
100, or 1000 times larger or smaller than the standard reference units and can be expressed using
common prefixes to denote the degree of difference (Table 2). Because of the base-10 system,
conversions within the metric system can be made relatively easily by either multiplying or dividing by
10.

When measuring with the metric system always use decimals (not fractions) to express your
measurements (e.g., 1.5 cm not 1 ½ cm).

Table 2. Prefixes for metric system units.

Prefix of Unit (Symbol) Part of Base Unit Value Example
kilo- (k) 1000 = 103 thousand 1 km = 1000 m
deci- (d) 0.1 = 10-1 tenth 1 dm = 0.1 m
centi- (c) 0.01 = 10-2 hundredth 1 cm = 0.01 m
milli- (m) 0.001 = 10-3 thousandth 1 mm = 0.001 m
micro- (µ) 0.000001 = 10-6 millionth 1 µm = 0.000001 m

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Examples of Converting Between Units

You can easily convert between two metric units by moving the decimal point according to how many
powers of 10 exist between the units. For each power of 10 by which the units differ, you shift the
decimal place one place. The superscripts in Table 2 indicate how many powers of 10 (how many
decimal places) separate base units from the units that include a prefix. For example, three decimal
places will convert any value of km to m or m to km. And three decimal places will also convert any
value of mm to m or m to mm.

Given the similarity, how do you know which way to move the decimal, to the right or to the left? The
key is to know which units are larger or smaller than the other unit.

When converting from larger units to smaller units, shift decimal to the right:
2.8 km = 2800.0 m = 280000.0 cm
[3 powers of 10 between km and m; 2 powers of 10 between m and cm]

When converting from smaller units to larger units, shift decimal to the left:
36.0 mm = 0.036 m = 0.000036 km
[3 powers of 10 between mm and m; 3 powers of 10 between m and km]

Another way to convert between units of length (or mass, or volume) is by using equivalent fractions.

In the example below, we convert meters to millimeters:

Convert 23 m to the equivalent length in mm, i.e. 23 m = ______?_____ mm

You know there are 1000 mm in every m, so you either divide the given “23” by 1000 or multiply 23
by 1000. Which is it? The fraction method makes it easy.

___23 m__ x 1000 mm = 23,000 (m x mm) = 23,000 mm = 23,000 mm

1 1m 1m 1

What The equivalent Multiply across the top and across the bottom,
you’re fraction with desired cancel the unit “m” that appears in both top
given over 1 unit in the top and the and bottom, simplify by removing the
unit you are converting denominator “1”.
from in the bottom.
Values for some
common conversions
can be found in Table 4
(pg. 8) of this handout.
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Activity I: Metric Measurement of Length
The standard unit of length in the metric system is the meter (m). In laboratory settings, the most
commonly derived units are the millimeter (mm) and centimeter (cm). For larger distances - such as
when driving a car - the kilometer (km) is used.

Length is measured with a ruler or meter stick. Intervals are marked in centimeter and millimeter
divisions. In the United States, rulers also typically include scale marks for Imperial intervals (inches)
separated into eighth and quarter divisions.

Figure 1. A comparison of imperial (inches; top) and metric (centimeters; bottom)

measurements. The metric ruler is further subdivided into millimeters; there are 10 small
marks (mm) per one cm.

The American Standard System (or Imperial System) includes units such as inches, feet, yards, and
pounds. Unlike the metric system, the relationship between units is not as “easy” to compare. For
example, the relationship between the three most common units of length is below:
12 inches (in) = 1 foot (ft)
3 ft = 1 yard (yd)

PROCEDURE:

Use Table 2 and the conversion examples to convert between the following metric units of length:

A. Practice: How many millimeters are in 3.67 m?

3.67 m = ___________________ mm

B. Practice: How many kilometers are in 480.0 cm?

480.0 cm = ___________________ m = ___________________ km

C. Using the metric ruler, measure the width of this page to the nearest tenth of a centimeter (e.g.,
10.3 cm). Using this measurement, convert the page width from centimeters, to meters, to
kilometers.
Page width = _________________ cm = __________________ m = ____________________ km

D. Using the American standard ruler, measure the width of this page to the nearest half-inch. Using
this measurement, convert the page width from inches, to feet, to yards.
Page width = _________________ in = ___________________ ft = ____________________ yd
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Activity 2: Metric Measurement of Volume
The standard unit of volume (the space an object
occupies) in the metric system is the liter (L). In
laboratory settings the most commonly used units are
the milliliter (mL or ml) or microliter (µL).

In the case of a box or cube, volume is relatively easy

to calculate: multiply the height by the width by the
depth. The values are measured as lengths. As such,
volume is sometimes reported in terms of cubic
centimeters (cc). The amount of water contained in a
cube with sides 1 cm long is 1 cubic centimeter (cc),
which equals 1 mL and has a mass of 1 g. However,
measuring non-standard volumes often requires
additional steps.

To measure the volume of a liquid, the liquid is

usually transferred into glassware such as beakers,
Erlenmeyer flasks, cylinders, or pipettes. This
glassware may be etched with measurement marks
(e.g., a graduated cylinder) which are typically
calibrated in mL. The top of the liquid forms a curved
surface known as a meniscus, and the volume of
Figure 2. Illustration of the proper way to
liquid is read at the lowest point of the meniscus
read the meniscus in a graduated cylinder to
(Figure 2). To assure accuracy in your measurements
measure the volume of a liquid.
you should always observe the meniscus at eye-level.

PROCEDURE:

Convert between the following metric units of volume. Consult Table 2 and the conversion examples as
needed:

A. Practice: How many milliliters and cubic centimeters are in 2.5 L?

2.5 L = ___________________ mL = ___________________ cc

B. Practice: How many liters are in 1.7 mL?

1.7 mL = ___________________ L

Practice using a graduated cylinder to determine the volume of water needed to fill a test tube:
1. Add water to a test tube until it is approximately half full.
2. Pour the water from the test tube into a 10-mL graduated cylinder and observe the meniscus to
determine the volume of water.

Volume of water = ___________________ mL

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Activity 3: Metric Measurement of Mass
Mass is the "matter" of an object and the standard metric unit used is the gram (g). Mass is measured
using a scale such as a triple-beam balance or electronic balance (Figure 3). It is important to note that in
a scientific context mass and weight are not synonymous. Weight is an object's mass multiplied by the
gravitational force applied to it. A person's mass would be the same regardless of if they were standing
on the Earth or the Moon, however their weight would be substantially different!

Figure 3. Illustration of an electronic balance.

As previously discussed in Activity 2, the amount of water

contained in a 1-cc cube would have a mass of 1 g. This means *+$$),-.
!"#$%&' ( ) !
that water has a density of 1. However, the mass of other /012*"),*3.
materials depends on its density. The density of a substance is its !
mass divided by its volume. Figure 4. Density equation.

PROCEDURE:

Use Table 2 and the conversion examples to convert between the following metric units of mass:

A. Practice: How many grams are in 500 mg?

500 mg = ___________________ g

B. Practice: How many milligrams and kilograms are in 2.5 g?

2.5 g = ___________________ mg 2.5 g = ___________________ kg

C. Practice: Calculate the mass of 100 cc of water.

Remember that 1 mL water = 1 g (at room temperature)
100 cc of water = 100 mL of water = __________________ g = __________________ kg

D. 1 L of seawater is found to have a mass of 1.028 kg. Calculate the density of seawater by dividing
the mass (in g) by the volume (in mL). Write your answer on the end of lab worksheet (pg. 9).

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Use an electronic balance to measure the mass of an unknown.
Note: Always zero-out the balance prior to measuring an object.
1. Place an empty 250-mL beaker onto the weighing platform and record the mass (g).
2. Add tap water to
the 250-mL beaker Table 3. Measurement of mass of a random amount of tap
so that it is about water.
half full. Measure Object Mass (g)
the mass of the
combined beaker Beaker ½ filled with Tap Water
and water and Empty Beaker
record the result in
Table 3. Tap Water (by subtraction)
3. Subtract the mass
of the empty beaker from the mass of the combined beaker and water to determine the mass of the
water. Record this result in Table 3.
4. Once you have calculated the mass of the water, use a graduated cylinder to measure the volume
of water in milliliters.
Volume of tap water = __________________ mL

5. Record your measurements of the mass and the volume of the water as a fraction: g
mL
6. Based on your results, what is the density of tap water? Write your answer on the end of lab
worksheet (pg. 9).

Activity 4: Metric Measurement of Temperature

At a molecular level, temperature is the result of the movement of particles that make up an object or
environment. As the motion (and speed) of the particles increases the temperature also increases. Thus,
the degree of hotness or coldness is really a measure of molecular kinetics. This is typically measured
via a thermometer.

The standard unit of temperature in the metric system is the degree Celsius (°C). Like the other
components of the metric system, degrees Celsius also relies on a base 10 system. In Celsius, the boiling
point and freezing point of pure water at sea level differ by 2 powers of 10.

• The freezing point of water is 0°C

• The boiling point of water is 100°C
• The temperature range in between is divided into 100 equal units (each unit = 1°C)

In contrast to the metric scale of temperature, the American Standard scale [degrees Fahrenheit (°F)]
defines the freezing point of water as 32°F and the boiling point as 212°F. Fahrenheit was originally
derived using the average human body temperature. Because the scales differ in the size of the degree, a
change of one-degree Celsius represent a much larger temperature change than one-degree Fahrenheit; a

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change of 1.8°F corresponds to a change of 1°C. The only temperature at which both scales are the same
is -40°. The following equations can be used to compare Celsius and Fahrenheit temperatures:

Celsius (°C) to Fahrenheit (°F) Conversion: Fahrenheit (°F) to Celsius (°C) Conversion:
℉ = (℃ × 1.8) + 32° ℃ = (℉ − 32°)/1.8

PROCEDURE:

1. Using the equations above, calculate the following temperature conversions and include your
answers on the end of lab worksheet (pg. 9):
A. Room Temperature = 21°C B. Ice Water = 32°F
C. Boiling Water = 100°C D. Human Body = 98.6 °F

2. Use the provided thermometer to measure the ambient air temperature of the laboratory room.
When using the thermometer, avoid touching the red bulb end.

Laboratory Room Temperature = ___________________ °C

Table 4. Common metric conversions.

Length Mass Volume
1000000 µm = 1m 1000000 µg = 1g 1000000 µL = 1L
1000 mm = 1 m 1000 mg = 1g 1000 mL = 1 L
100 cm = 1 m 100 cg = 1g 100 cL = 1 L
10 mm = 1 cm 10 mg = 1 cg 10 mL = 1 cL
1000 m = 1 km 1000 g = 1kg 1000 L = 1 kL

Image Credits:
Figure 1: Metric and American Standard Ruler By EL Kasl
Figure 2: Graduated Cylinder with Meniscus By EL Kasl
Figure 3: Electronic Balance Illustration By EL Kasl

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BI 101 Lab Worksheet Name _________________________________ Section ______
Lab 1: Metric system

Activity 1: Metric Measurement of Length

1. 42.73 m = ___________________ mm
2. 1239.8 cm = ___________________ km

3. When converting the page width measurement, was it easier to calculate using the metric system or
the American Standard system? What characteristic(s) made your chosen method easier to convert?

Activity 2: Metric Measurement of Volume

* Assume water is at room temperature *

4. 0.693 L = ___________________ mL = ___________________ cc

5. 12.76 mL = ___________________ L
6. 6.2 µL = _________________ mL = __________________ L

Activity 3: Metric Measurement of Mass

7. 1497 mg = ___________________ g
8. 37.4 g = ___________________ kg
9. 540 cc of water = __________________ g = __________________ kg
10. Density of seawater according to your calculations = _________________ g/mL
11. Density of tap water according to your measurements = _________________ g/mL

Activity 4: Metric Measurement of Temperature

12a. Room Temperature = 21°C = __________________ °F

b. Ice Water = 32°F = __________________________ °C
c. Boiling Water = 100°C = ______________________ °F
d. Human Body Temperature = 98.6 °F = ___________ °C

13. True or False (circle): One degree on the Celsius scale represents a smaller change in temperature
than one degree on the Fahrenheit scale.

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14. At what temperature do the two systems “meet” (both have the same number for this temperature)?
________________ °C/°F

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BI 101 LAB 2
Scientific Method

Prepared by Dr. Emily Kasl, Dept. of Biology, UNA

OBJECTIVES
After completing these laboratory activities, you should understand / be able to:
• Explain the process of the scientific method.
• Identify and define the components of a controlled experiment: hypothesis, controlled variable,
dependent variable, independent variable, experimental group, and control group.
• Organize information to analyze your data and draw conclusions.
• Use the scientific method to solve real-world problems.

Introduction
The scientific method is a rigorous method of investigating phenomenon (Figure 1). The scientific
method generally starts with an initial observation of an interesting phenomenon. This observation can
then lead to the development of questions (e.g., why, or how does this phenomenon occur?) that require
further investigative research. When looking at new problems, scientists will often start by identifying
what is currently known about an issue before developing a hypothesis.

The hypothesis, a possible explanation for

the observation, is the basis for setting up
experiments. Essentially, scientists attempt
to answer their own question by devising a
testable explanation. A scientific
hypothesis is more than just an educated
guess or prediction. Rather, a good
hypothesis must be logical, based on sound
observation, and most importantly,
be testable. Once a hypothesis has been
constructed, a prediction is made: what
outcome would you expect based on the
hypothesis?

With that prediction in hand, the next step

of the scientific method is to design and
perform the experiment. Accurately
testing a hypothesis (so that you can draw
conclusions based on your results) relies
on good experimental design. Figure 1. The scientific method is not a step-by-step, linear
Experiments should be replicable, only process. Rather, it is a way to explore an observation or
one variable should be tested per experi- question based on testing and revising knowledge.
ment, and controls must be included.

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Once the experiment has been run, results are compiled from the data collected. Based on these results,
investigators can draw conclusions about the hypothesis. If the results support the prediction, then
the hypothesis is retained (supported). If the results do not support the prediction, then the hypothesis is
rejected (not supported). A hypothesis can be rejected but it can never be proven true. There is always
the possibility that the findings from additional experimentation or observation may no longer a support
a previously retained hypothesis. Investigators can then communicate their results to the scientific
community as well as broader society.

Activity I: Experimental Design

Variables are the measurable components of the experiment. The independent variable is the variable
that is changed or manipulated by the investigator during an experiment to produce a response.
The dependent variable is what is measured as a response to the independent variable. A given
experiment may have multiple dependent variables but should only have a single independent variable.

In controlled experiments, researchers designate testing groups that can be compared to each other. In
the experimental group, one factor (the independent variable) is changed. In the control group, the test
is performed under the same experimental conditions except there is no independent variable being
tested. By comparing the outcomes of these two groups, investigators can determine if the independent
variable is causing an effect.

In these controlled experiments, all other potential variables (other than the independent variable) are
held constant between both groups. The other variables not being tested can be referred to as controlled
variables or constants.

Real World Example: In a drug trial testing the efficacy of a new medication researchers will compare
the results of an experimental group (that receives the drug) and the control group (that receives a
placebo). In this example, the drug is the independent variable and the change that occurs because of the
drug is the dependent variable. To ensure the changes are only occurring due to the drug, all other
variables (such as diet, age, gender, and activity level) are tightly controlled (i.e., controlled variables).

PROCEDURE:

Consider the following three case studies and identify the components of their experimental design on
the end of lab worksheet (pg. 9):

Case Study 1: The Discovery of Penicillin

In 1928, Dr. Alexander Fleming was investigating the biology of Staphylococcus aureus, a common
bacteria on the surface of human skin that causes several diseases. After a summer break away from his
lab he returned, only to discover a few culture plates of S. aureus had been contaminated by a blue-green
mold. In plates where this mold, Penicillium, was present there were clear areas that contained no
bacteria. In culture plates without mold these clear areas were not present (Figure 2).

Given this observation, Fleming hypothesized that the mold must produce a chemical capable of killing
S. aureus. To prepare his experiment he first isolated the mold. Dr. Fleming then exposed the mold
isolates to a new culture of S. aureus. After exposure, Fleming observed that all the bacteria in the new
culture died. Having identified this as an anti-bacterial chemical, Fleming named the substance
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penicillin. However, it would take another decade before penicillin could be fully developed into the
wonder drug it came to be known as.

Figure 2. An illustration of two agar plates containing colonies of Staphylococcus aureus

(grey circles). In Plate A, the growth of the bacterial colonies has occurred unimpeded.
However, in Plate B, the presence of the blue-green Penicillium mold has led to a “halo”
wherein no bacteria can grow.

From the above description:

1a. What was Dr. Fleming investigating?

1b. What was Dr. Fleming’s hypothesis?

A hypothesis, by definition, must be testable. A common way to write a testable prediction, is using an
“IF” and “THEN” statement. Take, for example, the hypothesis: “the stove is hot.” A testable prediction
for this hypothesis would be “If I touch the stove, I will burn my hand.” The “IF” statement reflects the
independent variable (what is being tested; stove heat) and the “THEN” statement reflects the dependent
variable (the outcome; burning your hand).

1c. Rewrite Dr. Fleming’s prediction on the end of lab worksheet (pg. 9) in your own words, using
the if/then format.

1d. What was the independent variable for this experiment? What was the dependent variable?

1e. Do the results indicate that the hypothesis should be rejected or retained?
Explain your reasoning on the end of lab worksheet (pg. 9).

Case Study 2: Improving Plant Growth

You are interested starting a roadside produce business and wish to improve the quality of your peanut
plants. Nitrogen is an essential macronutrient for protein synthesis and photosynthesis in plants.
However, it is often considered to be a limiting nutrient – available in short supply in nature. In
agriculture, a common way to increase plant yield is to apply fertilizers that contain essential nutrients.

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You design an experiment that compares the height (growth) of two groups of plants after exposure to
fertilizer. Group A has 50 peanut plants that receive fertilizer. Group B has 50 peanut plants that do not
receive fertilizer. After two weeks you measure the changes in height and observe that plants from
Group A had an average height of 12.43 cm and plants from Group B had an average height of 12.38
cm.

2a. From this experiment, identify the following on the end of lab worksheet (pg. 9): the control
group, experimental group, independent variable, and dependent variable.

2b. Based on these results, what is your conclusion? How does fertilizer affect the growth of the
peanut plants?

Peanuts belong to the legume family and are noted for their association with symbiotic nitrogen-fixing
bacteria (Rhizobia). Living within the root nodules of the peanut plants, these bacteria can convert (or
fix) inorganic forms of nitrogen from the air and soil into a nitrogen source that is useable by the plant.
Because peanuts can easily obtain fixed nitrogen, they do not require additional nitrogen sources.

2c. Given this information, explain your experimental results on the end of lab worksheet (pg. 9).

2d. How could this experiment be improved considering the information above?

Case Study 3: New Drug Development – Positive & Negative Controls

Some controlled experiments may also incorporate positive and negative controls into the experimental
design. A positive control provides a baseline, or sets the standard, for what should happen if the
experiment is successful. On the other hand, a negative control is used to indicate what should happen
if the outcome of the experiment is unsuccessful (no change due to the tested variable).

Consider our previously introduced drug trial example. In this case, you are involved in developing a
new drug for the treatment of chronic headaches. To gain FDA approval for your drug, you designate
three trial groups each consisting of 100 individuals. Prior to the drug treatment, each participant was
monitored for a month and the number of headaches lasting more than 5 hours were recorded. Each
group was subsequently exposed to a month-long trial of one of three treatments:
Group A: Received a dosage of the new drug (Drug X).
Group B: Received a dosage of Naproxen (Aleve). This is a drug that is currently available for the
prevention and treatment of headaches.
Group C: Received a dosage of a placebo (sugar pill) that contains no medication.
After the trial period, the number of headaches reported by Group A decreased by an average of 3.5
headaches per month. Group B reported an average decrease of 3.2 headaches per month and Group C
reported an average increase of 0.3 headaches per month.

3a. On the end of lab worksheet (pg. 9) identify the independent variable, control group(s), and
experimental group.

3b. What is the importance of positive and negative controls within a scientific experiment?
3c. Based on these results, does Drug X appear to be a successful headache treatment? Explain.
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Activity 2: Performing an Experiment and Collecting Data
Cardiovascular fitness is a measurement of how well your body takes in oxygen and supplies that
oxygen throughout the body during sustained physical activity. A simple way to determine
cardiovascular fitness is by either measuring the heart rate or respiration rate of an individual after
prolonged physical exercise. A person who is more fit (has higher cardiovascular endurance) may have a
comparatively slower pulse or a lower respiratory rate after exercise.

The World Health Organization (WHO) suggests adults aged 18-64 years of age participate in at least
150 minutes of intentional exercise per week to avoid the negative health effects associated with a
sedentary lifestyle.1 Additionally, the type of aerobic exercise can have a different impact on
cardiovascular function; brisk walking, running, swimming, or even dancing is often associated with
more vigorous, heart-pumping results. In this exercise, you will work with your classmates to investigate
the impact activity level on cardiovascular fitness, as measured by heart rate.

PROCEDURE:

1. Students will be split into two groups and will participate in an activity for 60 seconds:
Group A. This will the low intensity activity group. Students will walk in place at a steady,
sustained rate.
Group B. This will be the high intensity activity group. Students will repeat a sit-stand motion.
Students sit on a chair with feet shoulder width apart and arms crossed across their chest. From this
position, the student will stand up (without pushing off with arms) and sit down repeatedly.

2. Prior to starting the experiment, we need to define the hypothesis:

If _______________________ exercise is associated with greater aerobic activity, then
(low intensity / high intensity)
the _____________________ activity group will have a greater change in heart rate after exercise.
(low intensity / high intensity)

3. Your instructor will act as timekeeper and

instructor you when to start and stop each
exercise. Prior to the exercise, you should
first practice taking your heart rate and to
find your resting heart rate.
• While sitting quietly use two fingers
(pointer and index fingers) to find the
pulse in the radial artery (Figure 3).
Figure 3. To take your pulse using the radial artery,
• On your instructor’s command, count place your pointer and index fingers between your
the number of beats per 15 seconds. wrist bone and tendon on the thumb side of your
Multiply this number by 4 to determine wrist. Press gently until you can feel each beat.
beats per minute (bpm).
Resting heart rate = ___________ bpm

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4. On your instructor’s command, begin your designated activity. After 60 seconds, stop and
immediately take your pulse following the procedure in Step 3.
Trial 1: Post-activity heart rate = __________________ bpm

5. Calculate your change in heart rate: Post-activity Heart Rate – Resting heart rate. Input your data
below as part of the class dataset in Table 1.
Trial 1: Change in heart rate = __________________ bpm

5. Wait at least 30 minutes before repeating Steps 3-5. On your instructor’s command, begin the same
designated activity as Trial 1. Input your data below and as a part of the class dataset in Table 1.
Trial 2: Post-activity heart rate = __________________ bpm
Trial 2: Change in heart rate = __________________ bpm

Table 1. Effect of activity level on pulse rate (bpm).

Change in Heart Rate (bpm)
Group A: Group B:
Student Low Intensity Activity High Intensity Activity
Trial 1 Trial 2 Trial 1 Trial 2
1
2
3
4
5
6
7
8
9
10
11
12
Average Change in
Heart Rate (bpm)

6. Calculate the average pulse rate for both low intensity activity and high intensity activity groups. To
find the average, take the total sum of all numbers (change in heart rate) for each activity group
and divide that sum by the number of data points collected (for example, 12 students = 12 data
points). Add your results to Table 1.

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 7. Use Table 1 to create a bar graph (Figure 4; pg. 10) comparing the average change in heart (pulse)
rates after low intensity activity and high intensity activity exercises. Answer the questions on the
end of lab worksheet (pg. 10).

Image Credits:
Figure 1: Adapted from: The Process of the Scientific Method by Jeremy Sato | CC-BY-NC-SA
[Link]
Figure 2: Penicillin Case Study Illustration by EL Kasl
Figure 3: Adapted from: Pulse Taking by Servier Medical Art | CC-BY-3.0-DEED ATTRIBUTION 3.0 UNPORTED
[Link]

1
World Health Organization. Guidelines on physical activity and sedentary behaviour. Geneva: World Health Organization; 2020.

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BI 101 Lab Worksheet Name _________________________________ Section ______
Lab 2: Scientific Method

Activity 1: Experimental Design

Case Study 1:
1c. Rewrite Dr. Fleming’s hypothesis using an “IF” and “THEN” prediction:

1d. In this experiment, the independent variable was ______________________________________

The dependent variable was _______________________________________________________

1e. Was Dr. Fleming’s hypothesis rejected or retained? Explain your reasoning:

Case Study 2:
2a. Group ________ was the control group. Group ________ was the experimental group.

What was the independent variable? ________________________________________________

What was the dependent variable? __________________________________________________

2c. What is a biological reason to explain the results of this experiment?

Case Study 3:
3a. Group ________ was the positive control group.

Group ________ was the negative control group.

Group ________ was the experimental group.

What was the independent variable? ________________________________________________

3b. What is the importance of positive and negative controls within a scientific experiment?

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Activity 2: Performing and Experiment and Collecting Data

4. In this experiment, what was the independent variable? __________________________________

5. What was the dependent variable? ___________________________________________________

Use the data from Activity 2 (Table 1) to make a bar graph. On the x-axis (horizontal axis) place the
independent variable and on the y-axis (vertical axis) place the dependent variable. Label both axes.

Figure 4. Effect of activity level on pulse rate (bpm).

6. Based on these results, was your hypothesis rejected or retained? Explain your reasoning.

7. Were there any weaknesses in this experimental design?

8. What additional variables could be controlled for?

9. What is an additional or modified hypothesis that you could test to determine other variables that may
impact cardiovascular fitness?

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BI 101 LAB 3
Biochemistry

Prepared by Ms. Rachael Nelson, Dept. of Biology, UNA

OBJECTIVES
After completing these laboratory activities, you should understand / be able to:
• Define the following terms: carbohydrate, lipid, protein, negative control, positive control,
indicator.
• Use Benedict’s solution, Lugol’s solution, Biuret reagent, and the brown paper test to determine
which macromolecules are found in each food sample.
• Contrast the use of positive and negative controls in the scientific method.

Introduction
Biochemistry is the study of the chemical aspects of molecules in living organisms. Life’s bodily
functions require macromolecules which are larger molecules comprised of smaller subunits or
monomers. The four major classes of macromolecules are carbohydrates, lipids, proteins, and nucleic
acids. Each class is comprised of organic compounds that contain carbon.

(A) Saturated Fatty Acid Chain (B) Lysine (C) Glucose

Figure 3.1 These three pictures show carbon forming the backbone of different compounds. (A)
Saturated fatty acid chains are used to build different classes of lipids. (B) Lysine is an amino acid
which are used to form proteins. (C) Glucose is a simple sugar used to form polysaccharides.

Indicators can be used to test for the presence of certain macromolecules in food samples. An indicator
is a chemical that causes a change from the initial color to show the presence of a certain organic
compound. Table 1 identifies the indicators that we will use in this lab to test for the presence of
specific organic compounds. We will not test for the presence of nucleic acids in this experiment.

Table 1. Indicators used to test for specific organic compounds.

Indicator Organic Compound
Brown Paper Test Lipids
Benedict’s Solution Simple Sugars
Lugol’s Solution Complex Carbohydrates
Biuret Reagent Proteins

1
When using an indicator, it is important to include a positive and negative control that will be used to
compare with your unknown results. A positive control establishes a baseline for the color change that
will occur when a certain organic compound is present in the sample. A list of the positive controls that
we will use in this laboratory can be found in the table below. A negative control establishes a baseline
for when a certain organic compound is absent in the sample. Distilled water is often used as a negative
control.

Table 2. Positive Controls used when testing for the presence of certain macromolecules.
Macromolecule Positive Control

Lipids Oil
Simple Sugars Glucose
Complex Sugars Starch
Proteins Albumin

Activity 1: Testing for the Presence of Lipids

Lipids are organic compounds that are insoluble and do not dissolve in water. Triglycerides, waxes,
phospholipids and steroids are included in this group of macromolecules. Lipids serve many functions
for the cell including long-term energy storage, structure and fluidity of the cell membrane, and cell
communication through hormones.
The brown paper test will be used to test for the presence of lipids. When a food sample that contains
lipids is placed on a brown paper towel, it will leave a greasy spot on the paper. Food samples that do
not contain lipids or have a very small amount will eventually evaporate from the brown paper bag and
not leave a greasy film.

PROCEDURE:
1. Draw 5 dime-sized circles on a sheet of brown paper towel with a pencil. Leave about 1-2
inches between each circle.
2. Write the name of the food sample under each circle as indicated in Table 3.
3. Stir the solutions with the appropriate pipet and then pipet a very small drop of each food sample
in the corresponding circle.
4. With a new paper towel, blot the drop.
5. After allowing time to dry, look for the presence of a greasy spot and record your results in Table
3. Enough time has passed when your distilled water drop has completely dried and is not
visible.
Table 3. Testing for the presence of lipids.
Circle Sample Presence of Greasy Area: Presence of Lipid:
Yes or No Yes or No
1 Distilled Water (negative control)
2 Oil (positive control)
3 Organic Peanut Butter
4 Processed Peanut Butter
5 Unknown
***Please refer to the worksheet to answer questions for Activity 1. ***

2
Activity 2: Testing for the Presence of Simple Sugars
Carbohydrates are organic compounds that provide short-term energy and structure and support for
cells. Monosaccharides are the monomer or building block of complex carbohydrates known as
polysaccharides. A few examples of monosaccharides are glucose and fructose. When two
monosaccharides bond together a disaccharide is formed. Monosaccharides and disaccharides are
commonly known as simple sugars.

Benedict’s solution will be the indicator used to test for the presence of simple sugars. If simple sugars
are present, the solution will turn to light green, yellow, orange, or dark red after being heated depending
on the amount of simple sugars present. A minimal amount of simple sugars is present if the solution
turns green and an abundant amount is present if the solution turns red.

PROCEDURE:
1. Fill a 400 mL beaker with 175 mL of tap water, and bring it to a gentle boil using the hot plate.
2. At the top of each test tube, use a permanent marker to label six test tubes with the corresponding
tube numbers as indicated in Table 4.
3. Use a ruler to measure 2 cm from the bottom of each test tube and draw a horizontal line with a
marker.
4. Fill each test tube with the corresponding solution sample to the line on the test tube.
5. Add 20 drops of Benedict’s solution to each test tube and gently swirl to mix.
6. Record the initial color of the solution (after adding Benedict’s solution) in Table 4.
7. Place the test tubes in the boiling water bath for one to three minutes.
8. Using the test tube holder, remove the test tubes from the water and place them back into the test
tube rack.
9. Record the final color of each test tube in Table 4, and fill in the last column based on your
results.

Table 4. Testing for the presence of simple sugars.

Tube Sample Initial Color Final Color after Presence of Simple
Boiled Sugars: Yes or No
1 Distilled Water
(negative control)
2 Glucose
(positive control)
3 Starch
4 Regular Soda
5 Diet Soda
6 Unknown

***Please refer to the worksheet to answer questions for Activity 2. ***

3
Activity 3: Testing for the Presence of Complex Carbohydrates
Complex carbohydrates are formed when many monosaccharides bond together to form
polysaccharides. Glycogen and starch are both complex carbohydrates that are used to store energy for
future use in animals and plants, respectively.

Lugol’s solution (iodine) will be the indicator used to test for the presence of complex carbohydrates. If
complex carbohydrates are present in the food sample, the solution will turn from amber to dark blue or
black.

PROCEDURE:

1. At the top of each test tube, use a permanent marker to label four test tubes with the
corresponding tube numbers as indicated in Table 5.
2. Use a ruler to measure 2 cm from the bottom of each test tube and draw a horizontal line with a
marker.
3. Fill each test tube with the corresponding sample to the line drawn on the test tube.
4. Record the initial color in Table 5.
5. Add 5 drops of Lugol’s solution to each test tube and gently swirl to mix.
6. Record the final color in Table 5 and fill in the last column based on your results.

Table 5. Testing for the presence of complex carbohydrates.

Tube Sample Initial Color Final Color after Presence or Absence
adding Lugol’s of Complex
Solution Carbohydrates: Yes
or No
1 Distilled Water
(negative control)
2 Starch
(positive control)
3 Glucose
4 Unknown

***Please refer to the worksheet to answer questions for Activity 3. ***

4
Activity 4: Testing for the Presence of Proteins
Proteins are organic compounds that contain long chains of amino acids. They serve many important
roles for living systems. For example, proteins can serve as catalysts which speed up chemical
reactions by lowering the activation energy. They also serve transport, regulatory, and structural roles
for cells.
Biuret’s reagent will be the indicator used to test for the presence of proteins. This reagent consists of
two solutions kept separately until time for testing. These solutions are potassium hydroxide and copper
sulfate. If proteins are present in the food sample, the solution will turn dark blue or purple.

PROCEDURE:

1. At the top of each test tube, use a permanent marker to label 5 test tubes with the corresponding
tube number as indicated in Table 6.
2. Use a ruler to measure 2 cm from the bottom of each test tube and draw a horizontal line with a
marker.
3. Fill each test tube with the corresponding sample to the line drawn on the test tube.
4. Record the initial color in Table 5.
5. Place five drops of each solution (potassium hydroxide and copper sulfate) into each of the test
tubes.
6. Place a square of parafilm over the test tubes and invert a few times.
7. Wait two minutes and then record the final color in Table 6 and fill in the last column based on
your results.

Table 6. Testing for the presence of proteins.

Tube Sample Initial Color Final Color after adding Presence of Proteins:
Biuret’s Reagent Yes or No
1 Distilled Water
2 Albumin (source
of protein)
3 Organic Peanut
Butter
4 Processed Peanut
Butter
5 Unknown

***Please refer to the worksheet to answer questions for Activity 4. ***

5
BI 101 Lab Worksheet: Biochemistry Name ___________________________ Section _______

Activity 1: Testing for the Presence of Lipids

1. Why are positive and negative controls important to include in an experiment?

2. When comparing the two types of peanut butter, did you see a difference in the lipid content?
Why or why not?

Activity 2: Testing for the Presence of Simple Sugars

3. When comparing the two types of soda, did you see a difference in the simple sugar content?
Why or Why not?

Activity 3: Testing for the Presence of Complex Carbohydrates

4. Explain why there is a difference in the results for starch and glucose

Activity 4: Testing for the Presence of Proteins

5. When comparing the two different types of peanut butter, did you see a difference in the protein
content? Why or why not?

6. Fill in the table based on your results for the unknown.

Macromolecules Presence: Yes or No
Lipids
Simple Sugars
Complex Carbohydrates
Proteins

7. The “unknown” represents the contents of an individual’s stomach. After reviewing three
different meal possibilities, form a hypothesis on which meal the “unknown” most likely
contains. Give evidence to support your hypothesis.

• Pie Factory Pizza- The individual loved to eat the Meat Lover’s pizza with pepperoni, sausage, and
bacon.
• Ricatoni’s Italian Grill- The individual loved to eat the famous bread and herbs with cheese ravioli in
a cheese sauce.
• Voodoo Wings- The individual loved to watch a sporting event and consume chicken wings and
celery.

Hypothesis:
_________________________________________________________________________
Evidence:
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________

6
BI 101 LAB 4
Microscopy and Cells
Prepared by Dr. Robin Whitekiller, Dept. of Biology, UNA
OBJECTIVES
After completing these laboratory activities, you should understand and be able to:
• Identify the parts of a compound light microscope as well as their functions.
• View specimens on the compound light microscope using the scanning, low, and high-power
objective lenses.
• Define terminology associated with microscopy: field of view, working distance, magnification,
resolution and depth of field.
• Prepare wet mounts of animal and plant cells.
• Identify (and provide functions) of organelles in plant and animal cells: cell membrane, nucleus, and
cytoplasm as well as organelles unique to plant cells: cell wall, chloroplasts, and central vacuole.
• Identify the parts of a dissecting microscope as well as their functions.
• View a specimen on the dissecting microscope.
• Briefly describe differences between the compound light and dissecting microscopes.

Introduction
Microscopes allow us to see specimens or parts of specimens that are too small to see with the naked
eye. Compound light microscopes use a light source or illuminator and objective lenses to magnify
objects up to 1000x. Dissecting microscopes are used to view larger specimens. They provide a three-
dimensional view of the specimens. There are also microscopes that provide greater magnification.
These microscopes use electrons instead of light. The transmission electron microscope allows us to
view details of structures too small to see with a light microscope. Scanning electron microscopes are
used to view the surface details of a specimen. In today’s lab, you will learn the parts of the compound
light and dissecting microscopes and how to use them to view specimens.

Activity 1: Learning the Parts of the Compound Light Microscope

To learn how to use the compound light microscope, we must first learn the parts. PROCEDURE: To
learn to the parts of the microscope, each student will need to move one closer. Do NOT slide them
across the tables. Pick one up and move it. One hand should be on the base with the other on the arm.
Do not tilt the microscope as the eyepiece may fall out. The microscope may already be plugged in.
Check and then unwind the cord at the back. Turn your microscope on using the switch found on the
back of your microscope. You should see a light coming from the light source or illuminator at the base
of the microscope. If not, you should adjust light intensity using the rheostat (red knob on the base).
Label Figure 1 (on the next page) with the following structures: ocular lens (eyepiece), nosepiece,
objective lens, mechanical stage, mechanical stage clips, mechanical stage knobs, iris diaphragm lever
(of the condenser), coarse focus knob, fine focus knob, rheostat, and the illuminator. Descriptions and
functions of the structures can be found in this paragraph and on the next two pages.

1
Figure 1. Compound Light Microscope.

Find the ocular lens or eyepiece. Our microscopes have one lens, but some compound light
microscopes have two. The ocular lens provides 10x magnification of a specimen being viewed. Next,
find the revolving nosepiece. It has three objectives lenses attached. Rotate the nosepiece and you will
see that each objective lens can be positioned over the stage opening. You should hear a clicking sound
as each lens locks into place. Always start with the scanning objective lens. It provides 4x
magnification. You can see the number written on the lens. The 10x objective lens (low-power lens)
magnifies a specimen 10x. The longest objective lens (the high-power lens) magnifies a specimen 40x.
In some courses, the microscopes will include a fourth lens (oil immersion lens). This lens can magnify
a specimen 100x.
To determine total magnification, multiply the magnifying power of the ocular lens times the power of
each objective lens. Magnification is the increase in size of the specimen image. PROCEDURE:
Determine total magnification for the low and high-power objective lenses using the scanning lens as an
example, and complete Table 1 (on the next page).

2
Table 1. Magnification table for the three objective lenses.
Objective Lens Magnifying Power Magnifying Power Total Magnification
of the Objective Lens of the Ocular Lens
Scanning 4x 10x 4x x 10x = 40x
Low Power 10x 10x 10x x 10x = x
High Power 40x 10x 40x x 10x = x

Locate the mechanical stage or stage. It is the large, square platform located just below the objective
lenses. The stage houses mechanical stage clips to hold a slide into place. The mechanical stage can be
moved using mechanical stage knobs. The top knob moves the stage forward and back while the lower
knob moves the stage left and right.

The condenser (located just beneath the stage) converges the light rays from the light source or
illuminator up through the stage and specimen. Can you see the light from the illuminator coming up
through the stage opening? If not, find your iris diaphragm lever. By moving the lever, the iris
diaphragm regulates the amount of light coming up through the condenser lens to the specimen on the
stage. Be sure the iris diaphragm lever is completely open before you proceed with the other activities.

Find the coarse and fine focus knobs. As you turn the coarse focus knob, watch the stage. It moves in
large increments up and down; therefore, it should only be used when viewing specimen with the 4x
objective lens. The fine focus knob moves the stage in very small increments and should be used with
the 4x, 10x and 40x objectives lenses to sharpen the image.

Activity 2: Additional Attributes of the Compound Light Microscope Demonstrated

with the Letter “e” Slide

PROCEDURE: Lower the stage all the way using the coarse focus knob. Obtain a letter “e” slide from
the slide box in your tray. Hold it by its edges to prevent smudging the slide with your fingerprints.
Before placing the slide into the stage clips, be sure it is oriented with the “e” as you would read it on
paper (i.e., right side up). If your slide has smudges, you lens paper to wipe it clean.

The scanning objective lens with 4x magnifying power should be in place over the letter “e”. Be sure the
lens clicks into place. Look from the side, not through the ocular. Using the mechanical stage knobs,
center the “e” so that it is directly over the stage opening. Using the coarse focus knob, raise the stage as
high as it will go. Looking through the ocular, slowly lower the stage using the coarse focus knob until
the letter “e” is clearly in view. You can also check to see if the fine focus knob will sharpen the image.
Notice that the letter appears inverted from top to bottom and left to right.

Rotate the low-power 10x objective lens into place over the slide. Notice that the distance between the
objective lens and specimen on the stage, the working distance, is greatly reduced. To focus the image,
ONLY use the fine focus knob. If you use the coarse focus knob with the longer lenses (10x and 40x),
you may damage the slide. You may need to adjust the contrast of the image with the iris diaphragm
lever.

Center the letter using the mechanical stage knobs. Note: when you move each knob, it moves the
specimen in the opposite direction. If you are trying to move the slide with the specimen forward while
looking through the ocular, the slide on the stage moves back toward you. If you try to move it to the left
3
while looking through the ocular, it moves to the right. As we increase magnification, the field of view
decreases; however, we see an increase in the resolution or the ability to distinguish details of a
specimen.

Now rotate the high-power 40x objective lens into place over the specimen. Use the fine focus knob to
sharpen the image. You may need to adjust the iris diaphragm or rheostat again. Depth of field or the
vertical depth of the image decreases as we increase magnification.
Answer the questions found in the Lab Worksheet.

Activity 3: Viewing Cells of Elodea, an Aquatic Plant

To see plant cells, we need to make a wet mount. PROCEDURE: Obtain a clean slide and cover slip.
Elodea can be found in a small bowl of water on the table. Using a pipette (plastic dropper) provided in
your tray, collect a small amount of water from the bowl that houses the Elodea. Place a single drop (or
two) of the water on the center of the slide by gently squeezing the bulb. Return the pipette to the bowl.
Obtain one leaf of the Elodea using the forceps (tweezers) in your tray. Place the leaf into the drop of
water. Obtain a cover slip (from another petri dish in your tray). These are small square plastic pieces
with a layer of paper on top. Remove the paper. Handle the coverslips from the edges to prevent
smudges. Gently place the coverslip onto the sample. You should see the water spread out under the
coverslip. If not, you may need to tap the coverslip lightly with the blunt end of the forceps. Wipe off
any excess water.

Place the prepared slide in the stage clips on the stage. Center the specimen. View your wet mount under
the 4x scanning objective lens. You should see many (very small) green rectangular-shaped cells
connected to each other. As we increase magnification, you should begin to see the individual cells and
their organelles more clearly. Shift to the 10x low-power lens. Only use the fine focus to improve the
resolution. Move the 40x high-power objective lens into place. Use the fine focus until some of the cell
details are more easily seen. Since we are seeing more than one layer of cells, you will need to adjust the
fine focus to see the top layer of cells. You may also need to adjust the lighting with the iris diaphragm
lever. After a while, you may notice green chloroplasts moving. This is cytoplasmic streaming, or
movement of the cytoplasm, a phenomenon created by the cytoskeleton presumably in response to
increased light and heat.

Plants have three structures not seen in animal cells. The cell wall, found outside the cell or plasma
membrane, is found in some protists (e.g., green algae) and all bacteria. Its function is to provide support
and shape to the cell. You can adjust the fine focus as well as the iris diaphragm. The large central
vacuole appears as a clear space within the cell. It is mostly liquid but contains some wastes. The central
vacuole helps to provide turgor pressure within the cell. It is the force inside the cell caused the fluid, or
water, that pushes the cell membrane against the cell wall. Chloroplasts are more easily visible because
they are green. Photosynthesis takes place in the chloroplasts. The nucleus is the control center of both
plant and animal cells. It houses the cell’s DNA and RNA. It is larger than the chloroplasts but not
always visible. Label the cell wall, central vacuole, and chloroplasts on Figure 2 (next page).

4
Figure 2. Elodea cells viewed using the high-power objective lens.

Answer the question found in the Lab Worksheet.

Activity 4: Viewing Cheek Cells (Animal Epithelial Cells)

To view our cheek cells using the compound light microscope, we will make another wet mount.
PROCEDURE: Get a paper towel from the stack on the table. Use the slide that you used to view
Elodea. Place the used Elodea in the trash and gently clean the slide using lens paper. Place your slide
on the paper towel. Obtain a fresh cover slip and place it next to the slide. Next, obtain a toothpick and
find the Lugol’s iodine stain in your tray. The iodine will stain your skin and clothes. Place a single drop
of the iodine on the center of your slide. The stain will increase your ability to see the cells and their
organelles against the background. Get the toothpick and GENTLY swab the inside of your cheek once
or twice to obtain epithelial cells. Stir the cells from the toothpick into the drop of iodine on the slide.
Immediately, take the used toothpick to the back of the room and place it into the red biohazard bag.
Do NOT lay it on the table!

Next, carefully take the coverslip and lay one edge just to the right of the stain. Slowly lower the
coverslip and you will see the stain (with your cells) spread. If you added too much iodine, gently
remove any excess using a piece of the lens paper. We do not want stain on the microscope. Place the
slide in the clips on the stage. Raise the stage all the way up. Center the specimen (looking from the
side). Looking through the ocular, slowly lower the stage until the cells are clearly in view. Observe the
cells with the scanning objective lens (4x). Since we used a stain, you will see very small yellow cells.
Once at least two or three cells are in focus, switch to the low-power objective lens (10x). Focus again
using only the fine focus. If the cells are clumped together, try to find some that are not. Once the cell or
cells are centered in your field of view, move the high-power objective lens (40x) into place and focus
(fine focus only). You may need to adjust the amount of light for the best resolution.

5
Label the nucleus (stained darker), the cytoplasm and the cell or plasma membrane on Figure 3 (below).
The cytoplasm is the fluid-like material found inside the cell but outside the nucleus. It houses
organelles or cell structures. The nucleus is the control center of the cell. It contains our genetic
material, DNA and RNA. It is surrounded by a nuclear membrane. The cell membrane surrounds
animal cells and regulates the movement of materials into and out of the cell. Once you are finished with
your observations, dispose of your slide and coverslip in the biohazard bag.

Figure 3. Cheek cells stained with iodine using the high-powered objective lens.
Answer the questions found in the Lab Worksheet.

Activity 5: Learning the Parts of a Dissecting Microscope

The dissecting microscope (also called stereomicroscope) is already set up on your table (see Fig. 4 on
the next page). You will work in pairs to learn how to use one. Lift and move your microscope to a place
where you and a partner can see it. The dissecting microscope is used to view specimens that are too
large and/or too opaque to be viewed on a compound light microscope. It is also useful for dissections.
There is a much greater working distance between the stage and the mechanical stage body, large
enough to place your hands on the sides of the stage to manipulate a specimen while at the same time
viewing the specimen.

6
Figure 4. Dissecting microscope.

PROCEDURE: Label Figure 4 with the following structures: ocular lens or eyepiece, objective lens,
magnification knobs (toggles between 1x and 3x magnification), focusing knob, stage plate, on/off knob
on the base with three types of illumination (I, T, and IT). There is a light intensity knob on side of the
base of the microscope opposite the on/off knob.

Answer the question found in the Lab Worksheet.

Activity 6: Learning to use the Dissecting Microscope

PROCEDURE: Be sure the microscope is plugged in and turn the on/off knob on the base of the
microscope on. The distance between the two ocular lenses can be adjusted to match the distance
between your eyes. Place the insect provided by your instructor on the stage plate (over the light source).
You do not need to remove it from the petri dish. You also do not need to use the stage clips. Switch the
on/off knob to I. Incident light (I) reflects off a specimen and is useful for seeing surface details. The
light intensity knob may be used with this type of light. Turn the knob to T. Transmitted light (T) passed
up through a specimen from below. You can also use both lights by switching to IT.
7
Find the small focus knob and check to make sure it says 1x (magnification). Since we also get 10x
magnification through the ocular lenses, the total magnification will be 10x. While looking through the
oculars, lower the microscope body (housing the objective lenses) using the large focusing knob until
you can see the specimen clearly. Once the image is clear, switch the small focus knob to 3x
(magnification). Total magnification is now 30X. You can use the large focus knob to sharpen the
image. The total magnification is much lower on the dissecting microscopes; however, this microscope
provides greater depth of field.

Draw the eyes, wings, mouthparts or antennae of the insect in the space provided in the Lab Worksheet.
Try the different light sources to see what works best.

Image Credits:

Compound Light Microscope By RR Whitekiller

Human Cheek Cells By Fritzmann |
[Link]
Elodea Cells By Juan Carlos Fonseca Mata | [Link]
_Microscopic_view_of_Elodea_canadensis.jpg
Dissecting microscope By RR Whitekiller

8
BI 101 Lab Worksheet: Microscopy and Cells Name ____________________ Section ___

Activity 2: Additional Attributes of the Compound Light Microscope Demonstrated with the Letter “e”
Slide

1. Define magnification.

2. What is the total magnification of the letter “e” using the scanning 4x objective lens?

3. What happened to the orientation of letter “e” when viewed through the ocular?

4. What is the total magnification using the low power 10x objective lens?

5. Define field of view. Does it increase decrease or remain the same as we increase magnification?

6. Define resolution. Does it increase, decrease or remain the same as we increase magnification?

7. Define working distance. Does it increase, decrease or remain the same as we increase magnification?

8. What is the total magnification of the letter “e” using the high-power objective (40x) lens?

9. Define depth of field. Does it increase, decrease or remain the same as we increase magnification?

Activity 3: Viewing Cells of Elodea, an Aquatic Plant

What are two structures seen in plant cells that animal cells do not possess, and what are their functions?
10.
11.

Activity 4: Viewing Cheek Cells (Animal Epithelial Cells)

12. What is the purpose of the Lugol’s iodine stain?

9
What are two organelles seen in animal cell and plant cells, and what are their functions?
13.
14.

Activity 5: Learning the Parts of a Dissecting Microscope

What are two benefits of using a dissecting microscope (when compared to using a compound light
microscope)?
15.
16.

Activity 6: Learning to use the Dissecting Microscope

17-18. Draw the eyes, a wing, mouthparts or antennae of the insect in the space below.

10
BI 101 LAB 5
Cell Membranes and Transport

Prepared by Mrs. Rachael Nelson, Dept. of Biology, UNA

OBJECTIVES
After completing these laboratory activities, you should understand / be able to:
• Define osmosis, diffusion, concentration gradient, and semipermeable membrane.
• Explain the concepts of osmosis and diffusion with the assistance of indicator solutions.
• Describe the effect of osmosis on living cells.
• Identify solutions as hypertonic, isotonic, and hypotonic based on a simulated cell’s percent mass
change over time.

Introduction
Living cells are enclosed by a plasma membrane consisting mostly of phospholipids and proteins
(Figure 1). The phospholipid bilayer of the cell membrane is semipermeable only allowing certain
molecules to pass through. Small, uncharged molecules can pass freely across the membrane. The
presence of proteins in the membrane allows additional molecules to pass from one side of the cell to
the other.

Figure 1. The plasma membrane consists of a phospholipid bilayer with the fatty acid tails oriented
towards the inside and the phosphate heads oriented towards the outside. Proteins are embedded
throughout the membrane.

A concentration gradient exists when there is a higher concentration of a certain molecule on one
side of the membrane compared to the other. As molecules move from an area of higher
concentration to an area of lower concentration, equilibrium is established through the process of
diffusion (Figure 2).
Due to the nature of cells consisting mostly of water, the process of osmosis is important in
maintaining proper water balance for the cell. Osmosis is the diffusion of water across a cell
membrane from an area of higher water concentration to an area of lower water concentration.

1
 Figure 2. Diffusion, over time, molecules diffuse from the extracellular fluid to the cytoplasm due to
random molecular motion until equilibrium is reached.

Activity 1: Diffusion of Solutes

The internal and external cellular environments consist of solutes, such as glucose and sodium ions,
dissolved in an aqueous solution where the solvent is water. The concentration of these solutes often
differs between the cell’s internal and external environment creating a concentration gradient. As a
mechanism of membrane transport certain molecules will diffuse across the membrane from an area
of high concentration to low concentration. In this activity, dialysis tubing will serve as the cell
membrane. We will determine what type of molecules can freely pass from one side of the
membrane to the other. The dialysis tubing contains microscopic pores that allows the passage of
solutes that are smaller in diameter than the pores. Glucose and starch will be used as solutes for this
activity. Remember glucose is a monosaccharide (a single sugar) and starch is a polysaccharide
(large molecule consisting of many glucose molecules bonded together).

Benedict’s solution will be used to test for the presence of glucose. If glucose is present, the blue
color will turn to green, orange, or red after being placed in a boiling water bath for 3-5 minutes.
Lugol’s solution will be used to test for the presence of starch. If starch is present, the amber color
will turn blue/black without placing the tubes in a boiling water bath.

PROCEDURE:
1. Fill a 250 mL beaker with 100 mL of distilled water.
2. Obtain a piece of dialysis tubing from the large glass bowl in the center of your lab bench and
close off one end using a binder clip. Make sure to twist the dialysis tubing before clinching it
with the clip.
3. Slide your index finger and thumb back and forth on the other edge of the dialysis tubing until it
opens. Continue with this motion down the dialysis tubing until the rest has been opened.
4. Using the labelled pipets, dispense four pipets-full (~ 8 mL) of glucose solution and four pipets-
full (~8 mL) of starch solution into the dialysis tubing.
5. Use another binder clip to close the free-end of the dialysis tubing. Make sure to twist the
dialysis tubing before clinching it with the clip.
6. At the sink, carefully rinse the outside of the dialysis tubing to remove any glucose or starch
solution.
2
7. Place the dialysis tubing into the 250 mL beaker with distilled water using a glass stirring rod.
See Figure 3 for assistance.

Figure 3. Dialysis tubing set-up.

8. Let the dialysis tubing sit undisturbed for at least 30 minutes.

9. Use the following symbols to illustrate the contents of the beaker and dialysis tubing at the
beginning of the experiment in the “Before Diffusion” diagram of Figure 5.
Starch - S
Glucose – G
Water - W

At this time, please proceed to set up Activity II and then return to complete Activity I.

10. Lugol’s solution (iodine) and Benedict’s solution will be used as indicators to determine if the
glucose and/or starch was able to pass the dialysis tubing into the beaker. Label four, new test
tubes as shown in Figure 4. The codes for labeling are defined below:

a. DT- in dialysis tubing I

I
b. BE- in beaker
c. B- Benedict’s solution
d. I- Lugol’s/Iodine Solution

Figure 4. Labeling test tubes for Activity 1.

11. Use a clean pipet to transfer one pipet-full of liquid from the dialysis tubing into the two test
tubes labeled, “DT”.
12. Use a clean pipet to transfer one pipet-full of liquid from the beaker into the two test tubes
labeled, “BE”.
13. Add 10 drops of Benedict’s solution to the two test tubes labeled, “DT/B” and “BE/B”.

3
 14. Place the two test tubes labeled, “B” into the boiling water bath for 1-3 minutes. Record the
results in Table 1.
15. Add 3 drops of Lugol’s solution (iodine) to the two test tubes labeled, “DT/I” and “BE/I”.
Record the results in Table 1.
16. Use the following symbols to illustrate the contents of the beaker and dialysis tubing at the end
of the experiment in the “After Diffusion” diagram of Figure 5.
Starch - S
Glucose – G
Water - W

Table 1. Diffusion results across dialysis tubing.

Test Tube Final Color (and heating if Positive (+) or Negative (-) Positive (+) or Negative
needed) for Starch (-) for Glucose
DT/B ---------------------------------
DT/I -----------------------------
BE/B ---------------------------------
BE/I -----------------------------

Before Diffusion After Diffusion

Beaker Beaker

Dialysis Tubing Dialysis Tubing

Figure 5. Diffusion results across the dialysis tubing.

***Please refer to the worksheet to answer questions for Activity 1. ***

4
Activity 2: Effects of Hypertonic and Hypotonic Solutions on Osmosis
Osmosis occurs when there is a different concentration of water between the internal and external
environments of cells. When cells are placed in an environment that contains a higher concentration of
solute than the inside of the cell, this type of solution is referred to as hypertonic. Hypertonic solutions
cause water to diffuse out the cell until equilibrium is reached causing the cell to shrink in size. When
cells are placed in an environment that contains a lower concentration of solute than the inside of cell,
this type of solution is referred to as hypotonic. Hypotonic solutions cause water to diffuse into the cell
until equilibrium is reached causing the cell to increase in size. Water moves from a hypotonic region to
a hypertonic region. Isotonic solutions contain the equal amounts of solute compared to another
solution; therefore, osmosis occurs at equal rates not changing the size of the cell.

In this activity, you will use Orbeez beads to identify two unknown solutions as hypertonic or hypotonic.
The percent change in mass will be used to determine if the Orbeez beads lost or gained water over time.
Orbeez beads are used in this activity to represent a cell. They contain a membrane that separates the
internal contents from the external environment.

PROCEDURE:

1. Obtain fourteen Orbeez beads. Divide them into two sets of seven. One set will be used for
Solution A and the other set for Solution B.
2. Obtain the mass for one set of seven Orbeez beads for Solution A and record their mass in Table
2.
a. Plug the balance in and turn it on.
b. Place a paper muffin cup on the scale and after it determines its mass, press the zero/tare
button.
c. Place the set of seven Orbeez beads in the paper muffin cup while still on the balance.
The number given is the mass of the seven Orbeez beads.
3. Place them on a paper towel labeled” Solution A” and set aside.
4. Using the same method from above, obtain the mass of the other set of seven Orbeez beads for
Solution B on the balance and record their mass in Table 2. Place them on a paper towel labeled
“Solution B” and set aside.
5. Pour 20 mL of solution A into a 50 mL beaker labeled, “Solution A” and place the seven Orbeez
beads for Solution A into the beaker.
6. Pour 20 mL of Solution B into a 50 mL beaker labeled, “Solution B” and place the seven Orbeez
beads for Solution B into the beaker.
7. Allow the Orbeez beads to soak for 30 minutes. (Return to Activity 1.)
8. Remove the Orbeez beads from Solution A with a spoon and carefully blot them off with a paper
towel. Use the balance to determine their mass and record it in Table 2 under the Final Mass (g)
column. Make sure to place the paper muffin cup on the balance first and zero/tare its mass
before placing the beads on the balance.
9. Remove Orbeez beads from Solution B with a spoon and carefully blot them off with a paper
towel. Use the balance to determine their mass and record it in Table 2 under the Final Mass (g)
column. Make sure to place the paper muffin cup on the balance first and zero/tare its mass
before placing the beads on the balance.

5
 10. Calculate the percent change in mass for the Orbeez beads in Solution A and B using the formula
below. Record it in Table 2.

Percent Change in Mass = {(Final Mass -Initial Mass)/Initial Mass} x 100

11. Determine if each solution is hypertonic or hypotonic by analyzing the percent change in mass
calculations and record your results in Table 2.

Table 2. Osmosis in Orbeez beads.

Solution Initial Mass Final Mass Percent Change in Type of Solution
(g) (g) Mass (%) (Hypertonic or
Hypotonic)
Solution A
Solution B

***Please refer to the worksheet to answer questions for Activity 2. ***

6
BI 101 Lab Worksheet: Cell Membranes Name __________________________ Section _______

Activity 1: Diffusion of Solutes

1. Explain why the cell membrane is semipermeable.

2. Using Table 1, form a hypothesis identifying which molecule(s) were able to diffuse across the
membrane.

3. Using Table 1, form a hypothesis identifying which molecules were NOT able to diffuse across
the membrane.

4. Explain the evidence for your hypothesis in number 2 and 3. (Hint- Use the results of your
indicator tests.)

5. Provide reasoning for your hypothesis. (Hint- Think about the size of the molecules.)

Activity 2: Effects of Hypertonic and Hypotonic Solutions on Maintaining Equilibrium in the Cell

6. What causes the increase in mass for the beads in solution A? Would solution A be considered
as a hypertonic, isotonic, or hypotonic solution?

7. What causes the decrease in mass for the beads in solution B? Would solution B be considered
as a hypertonic, isotonic, or hypotonic solution?

8. What would happen to the mass of a cell if it was placed in an isotonic solution?

9. Briefly explain how you identified the solutions as hypertonic or hypotonic.

10. Extension Question: A patient receives an IV infusion at the hospital. What would happen if
the IV infusion contained distilled water?

7
BI 101 LAB 6
Enzyme Activity

Prepared by Dr. Emily Kasl, Dept. of Biology, UNA

OBJECTIVES
After completing these laboratory activities, you should understand / be able to:
• Understand the function of enzymes in biological systems.
• Define the following terms in relation to enzymes and enzyme activity: substrate, product, active
site, catalyst, activation energy, and denature
• Interpret data in table and graphs to draw conclusions supported by experimental data.
• Explain how environmental factors (temperature, pH, concentration) can affect enzyme function.

Introduction
Cells are responsible for carrying out the metabolic
processes that make life possible. These chemical
reactions require a minimum amount of energy
(the activation energy) to spur the process into action.
Activation energy can be defined as the amount of
energy required to bring about a chemical reaction. In
most cases, the speed of these reactions is limited by the
activation energy; under normal conditions these cellular
processes would occur too slowly to sustain life.

To meet the rate of reaction necessary to sustain

life, enzymes are utilized. Enzymes, which are typically
proteins, speed up chemical reactions by lowering the Figure 1. Enzymes lower the activation
activation energy. Enzymes catalyze (change the rate of) energy of a reaction without changing
reactions by binding to and reorienting the substrate the overall energy released (ΔG).
molecules (reactants) in a way that increases the likelihood
that the reaction will proceed (Figure 1). The binding site is referred to as the active site, which is
uniquely shaped in association with specific substrates, like a lock (active site) and key (substrate)
(Figure 2).

Figure 2. Diagram illustrating a substrate binding to the active site of an enzyme.

1
Enzymes have several notable characteristics:
• Enzymes are not permanently changed or "used up" by the reactions.
• Enzymes are specific, with most enzymes associated with one particular type of reactant
(substrate).
• The function of an enzyme is dependent on its structure; changes to the shape of an enzyme can
lead to loss of function.

As with all proteins, enzymes are particularly susceptible to denaturation due to their complex, three-
dimensional molecular structure. When a protein becomes denatured (losing its shape) it loses its
activity. Enzymes are primed to function under conditions of homeostasis (a stable internal
environment) and large fluctuations against the homeostatic equilibrium can often greatly affect the
ability for cells to carry out the necessary functions of life.

The Enzyme Catalase

Catalase is one of the most common enzymes found in living cells of nearly all organisms. We will be
using the enzyme catalase to investigate the effects of environmental changes on enzymatic activity.
Cells use catalase to breakdown hydrogen peroxide (H2O2), a toxic substance that is a common
byproduct of metabolic reactions, into water and oxygen.
#$%$&$'(
Catalase catalyzes the reaction: 2 H! O! %⎯⎯⎯⎯⎯' 2H! O + O!

For this lab, we will be using catalase enzyme extracted from a potato and measuring catalase activity
indirectly. When hydrogen peroxide (substrate) is added to the potato extract (catalase enzyme) it will
generate visible foam (gaseous O2) as the reaction proceeds. After the reactions are over, we will
measure the height of the foam column as a way of determining how active the enzyme was for the
tested conditions.

Before we test specific environmental conditions (Activities 2 – 4), we need to observe the effect of the
enzyme when it is exposed to substrate (positive control) and when it is not exposed to substrate
(negative control). This will allow us to develop a measure of enzyme activity.

Activity I: Measuring Enzyme Activity

Note: In preparation of Activity 2, fill a 250 mL beaker
halfway with tap water and heat it to boiling on a hot
plate. Once your beaker is placed on the hot plate you
may continue with Activity 1.

PROCEDURE:

1. Label one test tube with a plus sign (+) and one
with a negative (-) sign. These will be your
positive and negative controls, respectively. On
each of these tubes draw a horizontal line 1 cm up
from the bottom of the test tube and another line
4 cm from the bottom of the test tube (Figure 3). Figure 3. Test tube measurement markings.

2
 2. Using a pipette, measure 1 mL of potato extract (fill the pipette to just below the bulb) and
dispense into the positive (+) test tube.
3. Into the positive control tube, add 3 mL of H2O2. Allow the reaction to proceed for 1 minute. After
1 minute, use a ruler to measure the height of the foam (in cm) from the 4 cm sharpie line (Figure
3).
Positive control foam height = _________________ cm
4. Using a pipette, measure 1 mL of potato extract (fill the pipette to just below the bulb) and
dispense into the negative (-) test tube.
5. Into the negative control, add 3 mL of distilled water. Allow the reaction to proceed for 1 minute
before using a ruler to measure the height of the foam (in cm) from the 4 cm sharpie line (Figure 3).
Negative control foam height = _________________ cm
6. The amount of foam can tell us something about the amount of enzyme activity. Remember: the
foam (gaseous O2) is produced as a product during the breakdown of hydrogen peroxide. Our
positive control should have a substantial column of foam. The number of cm of foam is our
measure of enzyme activity. Our negative control should have no foam (0 cm) thus an enzyme
activity level of 0.

For the remainder of our activities, the length of the column of foam (in cm) will be the measure of
enzyme activity.

Activity 2: Effect of Temperature on Catalase Activity

The ability of temperature to affect the function of proteins (including enzymes) is well known. The
ideal temperature for enzyme function is typically associated with the homeostatic ideal of a given
organism. For instance, humans have an internal body temperature of 37oC (98.6oF). Thus, the enzymes
in our body are primed to function best at 37oC. In these experiments, the catalase is derived from
potatoes, which have a very different ideal temperature range.

Before we begin this activity, quickly recap the design of this experiment:

A. What is the independent variable tested in Activity 2? ______________________________

B. What is the dependent variable tested in Activity 2? ________________________________

PROCEDURE:

1. Set up the following temperatures:

• Fill a beaker with ice obtained from a Styrofoam cooler. This will be your 4ºC temperature bath.
• Grab an empty test tube rack and place at your station. Use a thermometer to record the ambient
temperature of the lab room. Typically, this is between 20-22ºC (68-72ºF).
Room Temperature (RT) = ___________ ºC
• Once your beaker atop the hot plate reaches a rolling boil you can turn down the temperature
slightly. This will be your 100ºC temperature bath.
3
 2. Take three clean test tubes and label them 4-E, RT-E, and 100-E. These labels represent the
temperature they will be placed in (e.g., 4ºC) and that the test tube will contain the enzyme. As in
Activity 1, draw a sharpie line at 1 cm and 4 cm from the bottom of the test tube (Figure 3). Into
each test tube place 1mL of potato extract (catalase). Use a sharpie to draw a line on your test tube
to mark the level of solution.
3. Take three clean test tubes and label them 4-HP, RT-HP, and 100-HP. Into each test tube place 3
mL of hydrogen peroxide. Set aside.
4. In pairs [e.g., 4-E and 4-HP] put the test tubes into the appropriate water baths and test-tube racks.
Do not mix the potato extract and hydrogen peroxide yet. There should be two test tubes (an -E and
a -HP) at each temperature. Allow the test tubes to sit in each temperature for at least 4 minutes.
5. After 4 minutes, use the test tube holder to remove the tubes from the boiling water and place them
in the room temperature test tube holder. Allow these two test tubes to cool to room temperature
over approximately 5 minutes (should be cool to the touch) before completing step 6.
6. Add the tube of hydrogen peroxide (-HP) into its corresponding tube of potato extract (-E; same
temperature). Start with the ice and room temperature test tubes first, immediately after removing
them from their respective temperatures. Do not test the 100ºC test tubes until they come to room
temperature. Allow the reaction to proceed for 1 minute. After 1 minute, use a ruler to measure the
height of the foam (in cm) from the 4 cm sharpie line. Record these results in Table 1.

Table 1. Effect of temperature on catalase activity.

Temperature Foam Height (cm) Observations
4ºC
RT = _____ºC
100ºC

Use this table to graph your data and answer questions on the end of lab worksheet (pg. 7).

Activity 3: Effect of pH on Catalase Activity

Much like temperature, pH is also an important characteristic of a stable internal environment.
Organisms can be adapted to function best in neutral, acidic, or basic environments, depending on where
they are commonly found. However, the ideal pH within an organism can vary. Enzymes often have a
very narrow pH range that coincides with optimal function.

Consider a human example: pepsin, an enzyme which assists in the digestion of proteins, is found in the
stomach. The stomach is filled with very acidic hydrochloric acid. Thus, pepsin works best at an acidic
pH (~2). In contrast, trypsin, a digestive enzyme found in the intestine (pH ~7), works best at a neutral
pH.

Before we begin this activity, quickly recap the design of this experiment:

A. What is the independent variable tested in Activity 3? ______________________________

B. What is the dependent variable tested in Activity 3? ________________________________

4
 C. Predictions: based on the results of Activity 2, how do you think pH may affect the activity of
catalase? Prior to starting the experiment, write down your predicted enzyme activity levels for a
pH of 3 (acidic), 7 (neutral), and 11 (basic) in Table 2.

PROCEDURE:

1. Label 3 test tubes “pH 3,” “pH 7,” and “pH 11.” On each test tube draw two sharpie lines at 1 cm
and 6 cm from the bottom.
2. Add 1 mL of potato extract (catalase) into each tube.
3. Add 2 mL of the appropriate pH solution to the coinciding test tube.
4. Cover each test tube with parafilm and thoroughly mix the contents by flipping the test tube upside-
down and right-side up for 5 seconds.
5. Place three test tubes in a test tube rack at room temperature. Wait for 10 minutes.

Note: While waiting, set a timer and continue to Activity 4. After 10 minutes, resume with step 6.

6. Add 3 mL of hydrogen peroxide to each test tube. If necessary, cover test tube with parafilm and
agitate slightly to mix. Allow the reaction to proceed for 30 seconds. After 30 seconds use a ruler to
measure the height of the foam (in cm) from the 6 cm sharpie line. Record these results in Table 2.

Table 2. Effect of pH on catalase activity.

pH Predicted Foam Height (cm) Foam Height (cm) Observations
3
7
11

Use this table to graph your data and answer questions on the end of lab worksheet (pg. 7).

Activity 4: Effect of Concentration on Catalase Activity

Recall that enzymes, on a molecular level, are not permanently changed (or "used up") during a reaction.
Rather, enzymes are like a pair of scissors or a stapler, they operate multiple times converting substrates
to products. So long as there is substrate available for the enzyme to bind with, the reaction will proceed.
Enzyme concentration, or the amount of enzyme available to be utilized in a reaction, can be an
important driver of the overall rate of reaction. This experiment will review how the ratio of enzyme
concentration (amount) compared to substrate amount effects catalase activity. In addition to foam
amount, also pay attention to the differences in speed as these reactions occur.

Before we begin this activity, quickly recap the design of this experiment:

A. What is the independent variable tested in Activity 4? ______________________________

B. What is the dependent variable tested in Activity 4? ________________________________

5
PROCEDURE:

1. Take three clean test tubes and label them 1, 2, and 3.

Tube 1: Draw a sharpie line 3 cm from the bottom. Add 1 mL of potato extract into tube 1.
This is a 0.5:1 ratio [33% concentration] of enzyme to substrate.
Tube 2: Draw a sharpie line 4 cm from the bottom. Add 2 mL of potato extract into tube 2.
This is a 1:1 ratio [50% concentration] of enzyme to substrate.
Tube 3: Draw a sharpie line 5 cm from the bottom. Add 3 mL of potato extract into tube 3.
This is a 1.5:1 ratio [60% concentration] of enzyme to substrate.
2. Add 2 mL of hydrogen peroxide into each tube. Allow the reaction to proceed for 30 seconds or
until the foam “boils over” the top of the test tube. After 1 minute, use a ruler to measure the height
of the foam (in cm) from your previous sharpie line. If the foam extends past the test tube lip record
the activity as 5+. Record these results in Table 3.

Table 3. Effect of enzyme concentration on catalase activity.

Enzyme Concentration Foam Height (cm) Observations
33% enzyme (1 mL)
50% enzyme (2 mL)
60% enzyme (3 mL)

Use this table to graph your data and answer questions on the end of lab worksheet (pg. 8).

Image Credits:
Figure 1: Effect of Enzyme on Activation Energy | Biology 2e Fig 6.15 [Link]
Figure 2: Enzyme Binding | Biology 2e Fig 6.16 [Link]
Figure 3: Test Tube Illustration By EL Kasl

6
BI 101 Lab Worksheet Name _________________________________ Section ______
Lab 6: Enzyme Activity

Activity 1: Preparing Catalase Controls

1. In the reaction being studied, what is the substrate? _____________________________________

what is the enzyme? _____________________________________

Activity 2: Effect of Temperature on Catalase Activity

Use the data from Activity 2 (Table 1) to make a line graph. On the x-axis (horizontal axis) place the
independent variable and on the y-axis (vertical axis) place the dependent variable. Label the axes and
draw a line connecting the three data points.

Figure 5. Effect of temperature on catalase activity.

2. What is the temperature at which the catalase activity is the greatest? _______________________

3. What is the temperature at which catalase has the least amount of activity? ___________________

4. What is a biological explanation for why the potato catalase works best (and worst) at these
temperatures? Think about where this enzyme is being obtained from and where it naturally occurs.

Activity 3: Effect of pH on Catalase Activity

5. At what pH is catalase activity the greatest? ___________________________________________

6. How do your predicted values compare to the results of this experiment?

7
Use the data from Activity 3 (Table 2) to make a line graph. Label the axes and plot both predicted and
experimental values. Differentiate and label the predicted and experimental result lines.

Figure 6. Effect of pH on catalase activity.

Activity 4: Effect of Concentration on Catalase Activity

Use the data from Activity 4 (Table 3) to make a line graph. Label the axes and draw a connecting line.

Figure 7. Effect of enzyme concentration (%) on catalase activity.

7. In addition to differences in enzyme activity rates (Fig. 7), did you observe any difference in the
speed of the reaction as the enzyme concentration increased? Explain why the reaction rate may
differ.

8
BI 101 LAB 7
Respiration and Photosynthesis

Prepared by Ms. Rachael Nelson, Dept. of Biology, UNA

OBJECTIVES
After completing these laboratory activities, you should understand / be able to:
• define cellular respiration, ATP, aerobic respiration, anaerobic respiration, and photosynthesis.
• explain how carbon dioxide produced by germinating seeds can be detected.
• explain how we determined carbon dioxide was consumed by photosynthesis.
• explain how a change in pH can be used to detect the presence of photosynthesis and cellular
respiration.

Introduction
Living organisms rely on photosynthesis and cellular respiration, as two processes that transfer energy
from one form to another. These metabolic processes are coupled, meaning the form of energy
produced in one process (photosynthesis) is used to fuel the other process (respiration). During
photosynthesis, organisms use solar energy to produce food (e.g., glucose) from carbon dioxide and
water. The chemical bonds present in glucose are then broken down through the process of cellular
respiration to form adenosine triphosphate (ATP), an energy molecule cells use to perform many of
their daily functions.

Activity 1: Creating an Indicator Solution

In this activity, you will be creating an indicator solution known as phenol red. The indicator solution
has the capability of changing colors as the pH of the solution changes. The phenol red solution is
red/pink under basic conditions (pH 8-14) and turns orange/yellow when it becomes more acidic (pH 0-
6). We will use this indicator for Activities 2 and 3 to explore cellular respiration and photosynthesis.
Figure 1 summarizes the color change that takes place.

Figure 1. The use of Phenol Red solution as an indicator for cellular respiration and photosynthesis.

1
PROCEDURE:

1. Obtain a 250 mL beaker and add 100 mL of tap water.

2. Place twenty drops of phenol red solution into the tap water. Gently mix with the glass stirring
rod.

Activity 2: Observing Carbon Dioxide Production of Germinating Seeds

Cellular respiration occurs when chemical energy from carbohydrates is transferred to ATP. If oxygen
is present in the cell, aerobic respiration will occur producing a relatively large yield of ATP, but if it
is absent, anaerobic respiration will occur producing a much smaller yield of ATP. Cellular
respiration occurs in both plants and animals. Animals obtain carbohydrates from consuming other
organisms, and plants produce glucose, a carbohydrate, through the process of photosynthesis. The ATP
produced from cellular respiration provides the required energy for many important metabolic processes
in the cells such as muscle contractions and active transport of molecules across the cell membrane.

Figure 2. The chemical equation for aerobic cellular respiration.

The generalized process (summary reaction) of aerobic cellular respiration is shown in Figure 2. To
determine if respiration is occurring, you can observe indirectly the production of carbon dioxide. In
Activity 2, you will compare the rate of cellular respiration occurring in germinated versus
ungerminated bean seeds. A germinated seed has begun to sprout and grow. Ungerminated seeds are
metabolically inactive, i.e. dormant. When carbon dioxide is produced, the pH drops because it
combines with water to produce carbonic acid (Figure 3). In the presence of the phenol red, a color
change should occur from red/pink to orange or yellow (Figure 1). The germinated bean seeds will be
enclosed in a test tube allowing the carbon dioxide to infiltrate the indicator solution.

CO2 + H2 O H2CO3
Carbon dioxide water carbonic acid

Figure 3. The chemical equation for the production of carbonic acid.

PROCEDURE:

1. Obtain two large test tubes labeled germinated and ungerminated beans. Using a pipet, transfer 6
mL of indicator solution into a graduated cylinder and pour it into the germinated beans test tube.
Repeat for the ungerminated beans test tube. See Figure 4 as a guide for set-up.
2. Using the large tweezers, push one cotton ball towards the bottom of the large test tubes, but not
touching the indicator solution. (approximately 2-3 cm above indicator solution)
3. Collect ten germinated beans from the side counter and place them in the test tube labeled,
“germinated”. The tweezers might be helpful in pushing the beans down. Place the stopper top
on the large test tube.
2
 4. Place ten ungerminated beans (from the bag at your station) in the test tube labeled,
“ungerminated”. Place the stopper on the large test tube.
5. Place both of the large test tubes into the test tube rack and record the color of indicator solution
every 15 minutes for the next hour.

Figure 4. Germinated and ungerminated bean apparatus.

Table 1. Results of Respiration in Germinated vs. Ungerminated Beans Experiment

Time (min) Ungerminated Beans Germinated Beans
(indicator solution color) (indicator solution color)
0
15
30
45
60

***Please refer to the worksheet to answer questions for Activity 2. ***

Activity 3: Absorption of Carbon Dioxide during Photosynthesis

Plants and some other organisms such as blue-green algae convert carbon dioxide and water into glucose
and oxygen through the process of photosynthesis. During this process, light energy is transformed into
chemical energy in the form of glucose. The generalized process (summary reaction) for photosynthesis
is shown in Figure 5.

3
Figure 5. Chemical equation for photosynthesis.

In this activity, you will use the indicator solution to demonstrate the occurrence of photosynthesis in a
plant sprig of Elodea. You will need to adapt your indicator solution to begin in an acidic form, so as
photosynthesis occurs and the plant uses the CO2 from the solution, the pH will rise resulting in a
red/pink color.

PROCEDURE:

1. Using a clean straw, carefully blow into the beaker of indicator solution until it turns from red to
orange-yellow. The carbon dioxide from your breath causes the pH of the solution to drop or
become more acidic.
2. Obtain the medium size test tubes labeled, “photosynthesis-exp and photosynthesis-con”. “Exp”
represents the experimental group, and “con” represents the control group.
3. Retrieve a full piece of Elodea (approximately 4.5 inches) and place it into the test tube labeled,
“photosynthesis-exp.”
4. Measure 25 mL of the acidified phenol red solution using a graduated cylinder and pour it into
the test tube labeled “photosynthesis-exp”. Please make sure the sprig has been completely
covered with the indicator solution. This will serve as your experimental group.
5. Measure 25 mL of the acidified phenol red solution using a graduated cylinder and pour it into
the test tube labeled “photosynthesis-con”. You will not add an Elodea sprig to this test tube.
This will serve as your control group.
6. Place a piece of Parafilm over each test tube to prevent the CO2 from escaping.
7. Place the test tube rack containing the two test tubes labeled, “photosynthesis-exp and
photosynthesis-con” in front of the light. Make sure the light is directed toward the test tubes.
Turn the light on.
8. If carbon dioxide is utilized by the Elodea, then the pH of the solution will increase (Figure 1).
The phenol red will return to its original color (red/pink) as it becomes more basic.

Table 2. Results of Carbon Dioxide Absorption Experiment

Time (min) Photosynthesis-experimental Photosynthesis-control
(indicator solution color) (indicator solution color)
0
15
30
45
60

***Please refer to the worksheet to answer questions for Activity 3. ***

4
Activity 4: Observing Cells of the Leaf Epidermis
As discussed in the previous activity, plants use photosynthesis to convert carbon dioxide and water into
glucose and oxygen. Oxygen and carbon dioxide are exchanged through tiny pores, known as stomata,
on the surface of the leaf. Water can also evaporate through the stomata in the process of transpiration.
The opening and closing of the stomata (stoma, singular) are regulated by guard cells found
surrounding the stomata. Figure 5 illustrates the underside of a leaf containing stomata and guard cells.

In Activity 4, we will use an ornamental house plant known as zebra plant, Tradescantia zebrina, to
observe the stomata and their guard cells on the underside of the leaf surface.

stomata

Guard cells

Epidermal cells

(a) (b)

Figure 5. Leaf Stomata (a) magnification of 40x (b) magnification of 100x

PROCEDURE:

1. Obtain a clean glass side from the petri dish. Obtain a small, square portion of a Trandescantia
zebrina leaf from your instructor and place it on the slide with the bottom portion facing up.
(The upper surface of the leaf has purple stripes while the lower surface of the leaf is uniformly
reddish-purple.)
2. Place a drop water from the glass container on the leaf and place a coverslip over it.
3. Place the glass slide onto the mechanical stage of a microscope. Use the scanning power
objective (4x) to bring the surface of the leaf into focus.
4. With the leaf in focus, rotate the low power objective (40X) in place to observe a total
magnification of 400X. Refocus and reposition the mechanical stage as needed to get a clear
view of the leaf’s surface features.
5. Use Figure 3 to identify the epidermal cells, guard cells, and stomata.

***Please refer to the worksheet to answer questions for Activity 4. ***

Image Credits:
Figure 1 image by RH Nelson
Figure 2 image by RH Nelson
Figure 3 image by RH Nelson
Figure 4 image by RH Nelson
Figure 5 image by RH Nelson
5
BI 101 Lab Worksheet: Respiration and Photosynthesis Name ___________________ Section ___

Activity 2: Measuring Carbon Dioxide Production of Germinating Seeds

1. What is the purpose of the tube containing ungerminated beans?

2. Explain why the indicator solution changed colors from red to orange/yellow in the test tube with
germinated beans?

3. How long (in minutes) did it take for carbon dioxide to be detected?

Activity 3: Absorption of Carbon Dioxide during Photosynthesis

4. Briefly explain why the pH of the phenol red was reduced prior to the activity.

5. Identify two constant/controlled variables between the experimental and control group in this
activity.

6. Why did the indicator solution turn from orange/yellow to red/pink in the tube with Elodea?

Activity 4: Observing Cells of the Leaf Epidermis

7-8. Draw a portion of the Zebra leaf as observed under the microscope and label the stomata and
guard cells.

9. As you observed the leaf’s surface, what cells contained chloroplasts? How could you tell these
were chloroplasts?

10. In most leaves the bottom layer contains many more stomata than the top. What benefit does this
provide for the organism?

7
BI 101 LAB 8
The Cell Cycle - Mitosis
Prepared by Dr. Robin Whitekiller, Dept. of Biology, UNA
OBJECTIVES
After completing these laboratory activities, you should understand and be able to:
• Provide some functions for the cell cycle.
• Define mitosis and cytoplasmic division or cytokinesis.
• Identify and describe cell cycle phases in both plant and animal cells.
• Determine the amount of time that cells spend in each cell cycle phase.

Introduction
Every day, our body cells divide and make copies of their DNA. The new cells must be identical to the
original cells to carry out the same functions. Cells go through the cell cycle to replace dead or damaged
cells (e.g., skin cells damaged by scrapes and cuts) and for growth and development. Some organisms
reproduce asexually (a mode of reproduction with a single parent). They rely on the cell cycle to
produce genetically identical offspring or clones.

Our chromosomes carry our DNA. Because we are eukaryotes, our chromosomes are located within the
nucleus of each body’s cells. The new cells produced must have the same number of chromosomes as
the original cell. In humans, our body cells have 46 chromosomes (23 pairs). For example, each new
skin cell produced should have 46 chromosomes.

The cell cycle includes interphase and mitosis. Mitosis is defined as nuclear division and includes the
following phases (in order): prophase, prometaphase, metaphase, anaphase, and telophase. Cytoplasmic
division (division of the cytoplasm) or cytokinesis is the final step of mitosis. The two new daughter
cells produced are identical to the parent cell except they are smaller with approximately 50% of the
cytoplasm and associated organelles. Each new cell produced will then begin the cell cycle again.

Cells spend the largest portion of the cell cycle in interphase. In interphase, the cell grows and prepares
for cell division. The chromosomes are also replicated or copied so that each new cell produced will
have an exact copy. The single-stranded chromosomes become double-stranded with two sister
chromatids joined via a centromere. The chromosomes are not visible during this portion of the cell
cycle but appear as chromatin, material that makes up the chromosomes. A nuclear membrane is
present and clearly visible. You may also see a nucleolus.

The first stage of mitosis is prophase. The chromosomes coil and condense so that they are clearly
visible. The nuclear membrane and nucleolus begin to break down and are not visible by
prometaphase. In the cytoplasm of animal cells, centriole pairs move toward opposite ends or poles of
the cell and form mitotic spindle fibers. In the absence of the nuclear membrane, the spindle fibers move
toward the double-stranded chromosomes. Plant cells do not possess centrioles.

1
The centriole pairs are seen at opposite poles of the cell during metaphase. We also see that the spindle
fibers have attached to the centromeres of each double-stranded chromosome. In both plant and animal
cells, the chromosomes line up along the midline or metaphasic plate of the cell.

During anaphase, the double-stranded chromosomes are pulled apart, and the sister chromatids of each
double-stranded chromosome begin to migrate toward opposite poles of the cell. At this point, the sister
chromatids become chromosomes once again.

Each group of single-stranded chromosomes reach opposite poles of the cell and become enclosed
within nuclear membranes during telophase. The chromosomes uncoil becoming invisible much like
they were during interphase. The mitotic spindle breaks down.

During cytoplasmic division or cytokinesis in animal cells, a cleavage furrow is visible as the cell
membrane begins to come together between the two nuclei. The result is the formation of two new
daughter cells. In plant cells, a cell plate made of cell wall chemicals forms between the two nuclei.
Once cytoplasmic division or cytokinesis is complete, the cell plate will become a component of the cell
wall of each new daughter cell and is responsible for dividing the parent cell in two. We should have
two new daughter cells with the same number of chromosomes as the original parent cell.

Figure 1 illustrates the cell cycle with five chromosomes in the original parent cell and five in each new
daughter cell. A drawing of prophase has not been included. PROCEDURE: Before you begin Activity
1, label the following structures in the figure below (in one phase): cell membrane, nuclear membrane,
nucleolus, chromatin, centriole pairs. double-stranded chromosomes, mitotic spindle fibers, single-
stranded chromosomes and cleavage furrow.

Figure 1. Cell cycle illustration with five chromosomes in an animal cell.

Activity 1: Modeling the Cell Cycle of an Animal Cell

To better understand how cell division works, we will begin by modeling the process for an animal cell
with five chromosomes. PROCEDURE: Work in pairs. There is a bag with all of the materials you
need in your tray. It include strings, straw pieces, pipe cleaners and pennies. Each string is labelled. The
longest string will represent a cell membrane. The second longest string will represent the nuclear
membrane, and the two shortest strings will represent the nuclear membranes for the daughter cells.
Four straw pieces will be used to represent the centriole pairs (two for each pair), but keep in mind that
plant cells do not possess centrioles. There are five different colored pipe cleaners to use as your
2
chromosomes. Each pair will vary in length. You will use two pipe cleaners of each color to represent
each double-stranded chromosome. One penny can be used to represent a nucleolus. Two are provided
for the daughter cells.

Interphase - Lay out the longest string to create a circle on the table to illustrate your cell membrane.
Add your nuclear membrane (second longest string). The chromosomes will not appear as double-
stranded until prophase. The material that you see is chromatin. However, the chromosomes are
replicated during interphase. Using two pipe cleaners of each color (and matching length), twist them
together at the center (once or twice) to represent the location of the centromere. Place all of your
double-stranded chromosomes in a pile in the nucleus. Include a penny in your model to represent a
nucleolus within the nucleus of the cell. Keep in mind that a plant cell would have an outer cell wall.

Prophase – Remove the nuclear membrane and nucleolus. Lay your double-stranded chromosomes out
so that they are clearly separate and distinct from one another.

Prometaphase – Centriole pairs (straw pieces) should be placed at opposite poles or ends of the cell
since this model represents an animal cell. No spindle fibers have been provided. Remember that plant
cells do not have centrioles.

Metaphase – Line up your double-stranded chromosomes along the midline or metaphasic plate of the
cell. They should be aligned between the centriole pairs. Mitotic spindle fibers attach to the centromere
of each double-stranded chromosome; however, the spindle fibers are not present in your model.

Anaphase – Gently untwist each double-stranded chromosomes and separate each sister chromatid.
Each chromatid begins to migrate towards opposite poles of the cell (towards the centriole pairs). They
are now referred to as chromosomes.

Telophase – Move the two sets of chromosomes to opposite poles of the cell. The animal cell develops
a cleavage furrow for cytoplasmic division. Demonstrate this by pulling together the cell membrane
along the center of the cell between the two nuclei. In plant cells, no cleavage furrow appears. Instead, a
cell plate will develop between the two nuclei. Find the two shortest strings (i.e., nuclear membranes)
and place one around each set of chromosomes. Each nucleus includes five single-stranded
chromosomes; however, they will uncoil and appear like the material you saw in interphase (before the
chromosomes were replicated). You can add a penny in each nucleus to represent the nucleoli.

Cytoplasmic division or cytokinesis – The cytoplasm pinches in completely creating a cell membrane
for each new animal cell produced. When you are finished, place all materials back in the bag and return
them to your tray.

Activity 2: Allium (Onion) Root Cell Cycle

We will use Allium a genus that includes onions to study the cell cycle in plants. PROCEDURE: Obtain
an onion mitosis (Allium root tip) slide from the box. There will be two to three onion root tip
longitudinal sections on the slide; therefore, if you do not find a stage on one, you can search the others.
Place your slide onto the stage. Looking through the ocular and using the coarse focus knob raise the
stage until one of the root tips is clearly in view. You can also use the fine focus knob. Using the
mechanical stage knob, move the root tip until the curved end is in view (see Figure 2 on page 3).

3
Figure 2. Allium (onion) root tip (400x magnification).

Center the root tip in your field of view. Rotate the 10x objective lens into place and use ONLY the fine
focus to sharpen the details. You should now notice that the cells are rectangular. Rotate the 40x
objective into place and use ONLY the fine focus to sharpen image details. Refer to Figure 3 for
assistance in identifying each phase. Once you have finished, leave the slide in place for the next
activity. If you have time before the next activity, move to Activity 4.

Interphase Prophase Prometaphase

Metaphase Anaphase Telophase

Figure 3. Allium (onion) root cell cycle phases (400x magnification).

4
Activity 3: Determine the Percent of Time Cells Spend in Each Phase of the Cell
Cycle

You may have noticed that most of the cells that you observed were in interphase. Cells spend the
majority of the life cycle in interphase. Now that you have learned to identify the phases of the cell cycle
in the plant cell, we will determine the percentage of time that these plant cells spend in each phase.
PROCEDURE: Under the 40x high power, scan the cells from left to right and top to bottom. Keep a
tally of the cells in each phase. Once you reach 100 cells, stop. Record your individual counts in Table 1
and provide your data to the instructor. Once everyone has finished, we will complete Table 1 with class
counts. We are increasing the accuracy of our counts by including class data.

To determine percent of cells in each phase, first determine the total number of cells counted in all five
phases using the class counts. Now determine the percent for each phase by taking the number of cells
seen in each phase and dividing that number by the total number of cells counted. Now multiply that
number by 100 to get the percent.

Table 1: Cell cycle phase counts.

Phase Individual Class Percent
Count Counts
Interphase
Prophase and Prometaphase
Metaphase
Anaphase
Telophase
Total 100 100

Based on the class data in Table 1, we can calculate how much time cells spend in each phase of the cell
cycle in a 24-hour period. First, change the percent to decimal form by moving the decimal two places to
the left. For example, 2% is equal to 0.02. Round to the nearest 1/100 (e.g., 0.02). Multiple 24 by the
number in decimal form. For example, 24 x 0.02 is equal to 0.48 and complete Table 2.

Table 2: Length of each cell cycle phase.

Phase Percent in decimal form Time (hours)
Interphase
Prophase and Prometaphase
Metaphase
Anaphase
Telophase
Total 1.00 24

Activity 4: Whitefish (Fish) Blastula Cell Cycle

To better understand the cell cycle in animal cells, we will view the stages of the cell cycle, in a
whitefish blastula. The blastula is one embryonic (developmental) stage seen in animals.
PROCEDURE: There are five microscopes set up around the room. Each microscope has a whitefish
blastula cross section in one of the cell cycle phases. Be sure you visit each microscope. Refer to Figure

5
4 for assistance in identifying each phase. If a cell has recently divided, you may see two smaller cells
side-by-side indicating cytoplasmic division or cytokinesis is complete.

Interphase Prometaphase Metaphase

Anaphase (early) Telophase (with cleavage

furrow)

Figure 4. Whitefish blastula cell cycle phases (400x magnification).

Image Credits:

Cell cycle illustrated By JM Marvin

Apical meristem in onion root tip By Wikimedia Commons
([Link]
Allium (onion) root cell cycle phases (400x magnification) By JM Marvin and RR Whitekiller
Whitefish blastula cell cycle phases (400x magnification) By JM Marvin and RR Whitekiller

6
BI 101 Lab Worksheet: The Cell Cycle-Mitosis Name ____________________ Section ___
______
What are two functions of the cell cycle? (page 1)
1.
2.

3. List the phases of the cell cycle in order. (pages 1 and 2)

4. Define mitosis. (page 1)

5. List the phases of mitosis in order. (page 1)

6. In which phase of the cell cycle do chromosomes line up along the midline or metaphasic plate of the
cell? (page 2)

7. In which phase of the cell cycle does each group of single-stranded chromosomes reach opposite
poles of the cell and become enclosed within nuclear membranes? (page 2)

8. In which phase of the cell cycle are chromosomes replicated? (page 1)

9. In which phase of the cell cycle do the double-stranded chromosomes first become visible after they
have coiled and condensed? (page 1)

10. In which phase of the cell cycle are the double-stranded chromosomes pulled apart? (page 2)

11. Based on Table 1 class data from Activity 3, what percent of time do cells spend in interphase?

12. Based on Table 2 class data from Activity 3, how much time do cells spend in interphase (in a 24-
hour period)?

13. In plant cells, what does the cell plate become following cytoplasmic division or cytokinesis? (page
2)

What are two differences between animal and plant cell cycles? [Hint: What do we see in plant cell
cycles that we do not see in animal cells? (pages 1 and 2)]
14.
15.

16. If a parent cell begins the cell cycle with 38 chromosomes, how many chromosomes should each
new daughter cell possess? (page 2)

7
BI 101 LAB 9
DNA - The Double Helix
Prepared by Dr. Robin Whitekiller, Dept. of Biology, UNA
OBJECTIVES
After completing these laboratory activities, you should understand and be able to:
• Describe the structure of DNA.
• Define key terms: DNA, nucleotide, DNA fingerprinting, gel electrophoresis, and polymerase chain
reaction.
• Construct a model of DNA.
• Extract DNA from plant tissue.
• Explain the process of DNA fingerprinting.
• Determine paternity of a child and solve a crime using DNA fingerprinting band sharing.

Introduction
DNA or deoxyribonucleic acid is a nucleic acid found in the nucleus of eukaryotic cells and the nucleoid
region of prokaryotic cells (e.g., bacteria). Each molecule of DNA is made of two nucleotide strands
bonded together and coiled into a double helix. Each nucleotide, the basic structural unit of nucleic
acids, is made of a nitrogen-containing base, a phosphate group and a five-carbon sugar (deoxyribose).
Each of four possible nucleotides differs in its nitrogen-containing base: adenine, thymine, guanine or
cytosine. The sugar-phosphate backbones of DNA are parallel to each other but run in opposite
directions. Since they run in opposite directions, they are considered antiparallel. Hydrogen bonds join
the two strands between specific nitrogen-containing bases. Two hydrogen bonds bind complementary
bases adenine and thymine, and three bind complementary bases guanine and cytosine. Watson and
Crick proposed the working model of DNA based on contributions by other scientists including
Franklin, Wilkins, and Chargaff.

Activity 1: Modeling the Structure of DNA

PROCEDURE: Work in pairs to construct a model. We will use a K’NEX DNA Replication and
Transcription Set to construct a DNA double-stranded molecule. Follow the instructions provided with
your set (pages 1-5). Your DNA model should include at least 10 nucleotides for each strand. Once you
have constructed the model, you will form the model into a double helix (pages 6 and 7). If you have
done everything correctly, your model should look like the image on page 7 (next to step 5). See Figures
1 and 2 on the next page for assistance. As your instructor to check your model before moving on.

1
 Figure 1. DNA, the molecular structure. Figure 2. DNA, the double helix.

Answer the questions found in the Lab Worksheet.

2
Activity 2: DNA Extraction

Strawberries will be used for this activity because they produce large amounts of DNA. We will use a
series of physical and chemical steps to “purify” the DNA from the cells of the strawberries.

PROCEDURE: Students will work in pairs.

1. Add one large strawberry or two small strawberries to a sealable plastic bag. Remove any air from the
bag, seal it and mash the strawberry or strawberries for three minutes to break open the cells. Do not to
leave any large chunks.

2. Open the bag and add four pipettes full of the DNA extraction solution (i.e., dishwashing soap and
salt). Close the bag and mix the strawberries with the solution for two minutes. The dishwashing soap in
the extraction solution will dissolve the cell and nuclear membranes to release the DNA. The salt
(sodium chloride) in the extraction solution will help neutralize the negative charge of DNA, making it
less soluble in water. The salt also removes some of the cellular proteins.

3. Add a piece of cheesecloth to a funnel, and insert the funnel into a beaker. Slowly, pour the contents
of the strawberry mixture from the bag into the funnel. The strawberry mixture will be filtered leaving
liquid in the bottom of the beaker. Fill a test tube approximately ½ full with the filtered liquid (i.e., the
filtrate).

4. Collect one pipette full of cold ethanol and add it slowly down the side of the test tube containing the
filtered liquid so that the ethanol does not mix with the filtrate. The stringy-looking DNA will become
visible between the ethanol and filtered liquid layers as it is less dense than the filtered liquid but more
dense than the ethanol (Figure 3).

5. Carefully, insert a glass rod below the layer of DNA and slowly pull some out.

Answer the questions found in the Lab Worksheet.

3
Activity 3: Using DNA Fingerprinting to Determine Paternity

DNA fingerprinting can be used to identify individuals since individuals (with the exception of
identical twins) have unique DNA patterns. Since we get 50% of our genes or alleles (forms of the
genes) from each parent, DNA fingerprinting can be used to determine relatedness between individuals.
It can also be used to identify suspects in criminal cases (e.g., murder) and to identify individuals who
cannot be identified through other means (e.g., a badly burned victim). To determine paternity (i.e., the
biological father of a child or offspring), a sample of blood is taken from potential fathers and the child.
DNA can also be extracted from other sources (e.g., saliva, semen, hair roots). The test is over 99%
accurate.

Refer to Figure 4 on the next page as we discuss the process of DNA fingerprinting. Once the DNA
is extracted (e.g., from a blood sample), it is cut into small fragments of different lengths (sizes) using
restriction enzymes. Restriction enzymes cut the DNA at specific sequences. The DNA fragments from
each individual are loaded into separate wells (small rectangular indentations) in an agarose gel using a
micropipette. An electric current is applied and the fragments of DNA are separated using gel
electrophoresis. An electric current will separate the fragments based on their size and charge. The
shortest fragments of DNA will travel the farthest (and fastest) in each lane. The electrical current
running through the gel causes the negatively charged DNA to migrate to the positive end. Next, the
fragments in the gel will be transferred to a nylon membrane using a vacuum blotter. The nylon
membrane will be incubated with a radioactive probe. Finally, the membrane will be exposed to X-ray
film (i.e., autoradiography). The fragments will appear as dark bands on the DNA fingerprint. Band-
sharing probabilities are used to determine relatedness. Remember that offspring share 50% of their
alleles with each parent. In some cases, there is not enough of a DNA sample. PCR or polymerase
chain reaction can be used to produce millions to billions of copies of a specific DNA segment, which
can then be used for fingerprinting.

Y-S
Figure 4: Steps involved in gel electrophoresis.

4
PROCEDURE: We will determine the paternity of the child (C) in the figure below. M = mother, F1 =
potential father 1, F2 = potential father 2, and F3 = potential father 3. In the space provided at the
bottom of the fingerprint, record number of bands shared by the mother and three potential fathers.
Answer the question found in the Lab Worksheet.

M C F1 F2 F3
___
___
___ ___
___ ___
___
___ ___
___
___
___ ___
___
___ ___

___ ___

Image Credits:

DNA, the molecular structure By Francescakb |

[Link]
DNA, the double helix By OpenStax College |
[Link] File:229 [Link]
DNA between the layer of ethanol and filtered liquid By RR Whitekiller
Steps involved in gel electrophoresis By Jenifer 0328 |
[Link]
Hypothetical DNA Fingerprints By RR Whitekiller

5
BI 101 Lab Worksheet: DNA - The Double Helix Name ____________________ Section ___

Activity 1: Modeling the DNA Structure

1. How many nucleotides are present in the DNA of Figure 1? [Note the rectangle drawn around one
nucleotide in the figure: S = deoxyribose sugar, P = phosphate group, and Guanine = nitrogen-
containing base]

2. If we look at nitrogen-containing bases on one short piece of a DNA strand, what would be the
sequence of the bases on the complementary strand? The first base on the complementary strand is
provided.

A T T G A C G A

3. What type of chemical bond joins the complimentary nitrogen-containing bases together between the
two DNA strands?

Activity 2: DNA Extraction

4. Why did we mash the strawberries before adding the extraction solution?

5. What is the purpose for the dishwashing detergent in the DNA extraction solution?

6. What are two purposes for the salt in the DNA extractions solution?
A.
B.

Activity 3: Using DNA Fingerprinting to Determine Paternity

Refer back to the bottom of page three and Figure 4 on page four to fill in the blanks below.
Once DNA is extracted, it is cut into fragments using _______________ _______________ (7). The
fragments are separated using _______ _______________ (8). The fragments are transferred to a nylon
membrane and incubated with a ________________ ______________ (9). The membrane will be then
be exposed to X-ray film in a process known as ____________________ (10). The DNA fragments will
appear as dark bands. If there is not enough of a DNA sample, _______________ __________
__________ (11). can be used to produce millions to billions of copies of a specific DNA segment.
12. Based on the DNA fingerprint results (i.e., band sharing) on page four, which individual is the
biological father of the child?
6
Gel electrophoresis separates DNA fragments based on their ___ and ___.
13.
14.
As stated previously, DNA can also used to solve crimes. Below, we have a DNA fingerprint from a
criminal case. Tissue was collected from underneath the victim’s nails. In the space provided below the
fingerprint, record number of bands shared by the tissue sample, the victim and the suspects.

Victim Tissue Sample Suspect 1 Suspect 2 Suspect 3

_____
_____ _____
_____
_____
_____ _____
_____

_____
_____
_____
_____ _____
_____
_____
_____
_____

15. In the DNA fingerprint above, which suspect’s DNA was found under the victim’s nails?

7
BI 101 LAB 10
Microevolution in Populations

Prepared by Ms. Rachael Nelson, Dept. of Biology, UNA

OBJECTIVES
After completing these laboratory activities, you should understand / be able to:
• Define population, microevolution, gene pool, genetic drift, bottleneck effect, founder effect, and
natural selection.
• Explain how random events can cause a change in a population’s gene pool.
• Explain how natural selection can cause a change in a population’s gene pool.
• Collect data, graph data, and draw conclusions based upon the data.

Introduction
Environments are home to many different populations of organisms. A population is a group of
organisms of the same species living in the same area. Environments are constantly changing over time,
and these environmental changes may favor different genetic traits within a population. Microevolution
is defined as the change in allele (different version of a gene) frequencies over time. We will look at
two events that cause microevolution: genetic drift and natural selection.

Natural selection is a non-random event in which the environment selects for a favored genetic trait such
as a particular fur color or the shape of a bird’s beak. Since this trait is selected for, these organisms are
more likely to survive and pass on these traits to their offspring. For example, if an environment faces a
severe drought, birds with larger beaks may be more likely to survive. They were able to crack open the
larger, thicker seeds to maintain energy for their daily lives. Through natural selection, the gene pool
(i.e., all the alleles available in the population), can shift favoring one allele over another. In the case of
frequent droughts, larger beak size is favored.

Genetic drift is a chance event in which the allele frequencies of a population can randomly change
from one generation to the next. Unlike natural selection, one trait is not favored over another. The
bottleneck effect and founder effect are two examples of genetic drift. Bottleneck effect happens when
there is a drastic reduction in the population size. For example, a tornado would reduce the size of a
population, but who survives is due to chance alone. Overhunting and disease are two other examples
that might lead to a bottleneck effect. The founder effect occurs when a small portion of a population
becomes separated from the main population and forms a new population. There is a reduction in the
genetic variation due to the small number of individuals in the new population. For example, a strip of
land separating a stream might cause a large population of fish to become two smaller populations.

In the following activities, you will analyze the change in gene pools in a population of artic hares with
brown or white fur. The first activity will simulate a population experiencing genetic drift. The second
and third activities will simulate a population experiencing natural selection. Lastly, you will graph the
data from all three activities to determine how genetic drift and natural selection affect the allele
frequencies.

1
Activity 1: Simulation of Genetic Drift with the Bottleneck Effect
Brown and white rabbit beads will be used to represent brown and white alleles present in a species of
artic hares. We will simulate some catastrophic event (e.g. a blizzard) to reduce the population size.
Chance alone will determine which individuals survive to produce offspring. Each lab bench will
determine if the population’s allelic frequencies of brown and white hares are changing over time.

PROCEDURE:

1. Place 20 brown and 20 white-colored hare beads in the plastic beaker, gently mix them, and
randomly place them on the tray. Be sure they are spread out. This will represent your starting
population (Generation One) of 40 hares that consists of 20 white hares and 20 brown hares.
2. Record the amount of brown and white hares for Generation One (Table 1).
3. A catastrophic event occurs and only four of the forty hares survive. Without looking, pick up four
hares and place them in the beaker. Remove the remaining 36 hares. (These 36 hares did not survive
the catastrophe.)
4. Record the numbers of surviving brown and white hares (in the beaker) for Generation One in Table
1.
5. The surviving hares reproduce to build the population size back to 40. (For example, if you had 3
brown hares and 1 white hare, multiply each color total by 10 to calculate the new starting
population size. In this example, “Generation Two” would start with 30 brown hares and 10 white
hares.)
6. Record the number of hares at the beginning of Generation Two in Table 1.
7. Replenish your brown and white hares in the tray to represent Generation Two and place the hares
not needed back in the original containers.
8. Repeat steps 3-7 to determine which hares survived Generation Two and started Generation Three.
Make sure to record your data in Table 1. Continue this process until all data has been collected
through Generation Five.
If fixation occurs, then your lab group can stop the simulation at that generation. Fixation occurs
if either the brown or white hare number reaches zero and the surviving color occurs in every
member of the population. One allele has been lost from the population and the other is fixed at
100%.
9. Record each lab group’s number of white hares at the beginning of each generation in Table 2.
These will be used to construct a graph in Activity 4.

Table 1. Lab group results from the simulation of population experiencing genetic drift (Activity 1).
Generation Number of Hares at the Beginning of Number of Surviving Hares
the Generation (Out of 4)
(Out of 40)
White Brown White Brown
One
Two
Three
Four
Five

2
Table 2. Class results from the simulation of population experiencing genetic drift (Activity 1).
Generation Number of White Hares at the Beginning of Each Generation
Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
One
Two
Three
Four
Five

Activity 2: Simulation of a Population Experiencing Natural Selection during the

Summer
In this activity, you will analyze the effects of natural selection. Natural selection is the differential
survival and reproduction of organisms due to heritable adaptive traits (i.e. traits that may lead to
survival and/or reproduction). The adaptive characteristic you will be investigating is cryptic coloration.
Cryptic coloration or camouflage is an antipredator mechanism used by organisms to avoid detection in
their environments. It may include coloration, shape and or behavior. Figure 1 provides two examples
of cryptic coloration.

You will simulate the predation of brown and white hares against a dark background. The dark
background will include a textured, brown, gray, and black fabric that represents the ground cover
during the summer months in the taiga. Each lab group will determine if the population’s allelic
frequencies of brown and white hares are changing over time.

Figure 1. (a) A walking stick with the same color and shape as the stem of this plant, and (b) the
chameleon use body shape and color to avoid predation. (OpenStax 2e)

3
PROCEDURE:

1. Have the two predators look away while the other partners place 20 brown and 20 white-colored
hares in the plastic beaker, gently mix them, and randomly place them on the dark textured
fabric where they are evenly spread out. Take a few minutes to fluff the fabric so the hares fall
in between the faux fur. This will represent your starting population (Generation One) of 40
hares that consists of 20 brown hares and 20 white hares.
2. Record the amount of brown and white hares in the beginning of Generation One (Table 3).
3. First round of predation:
a. One person will be the timer, two people will be predators, and the fourth person will be the
recorder.
b. The predators need to wear sunglasses and the instructor may choose to dim the lights before
predation begins.
c. When the timer says, “GO”, the predators will begin to feed for 10 seconds. The predators
will use their sight to capture one hare at a time and place it into a beaker. The predators
should capture prey as quickly as possible. After ten seconds, the timer says, “STOP”.
4. Record your lab group’s total “number of hares captured” (in the beakers) for each phenotype
(brown and white hares) for Generation One in Table 3.
5. Determine the “number surviving” brown and white hares using the formula below. Record
these values in Table 3.

# of surviving white hares = # of white hares at the beginning of the generation - # of total white hares
captured
# of surviving brown hares = # of brown hares at the beginning of the generation - # of total brown
hares captured

6. To build the population back up to 40, assuming the surviving hares produced offspring similar
to themselves:
a. Calculate the frequency of the surviving brown and white hares using the formula below.
Record the frequency of brown and white hares to two decimal places (e.g., you would round
0.456 to 0.46) in Table 3.

# of surviving white hares = frequency of surviving white hares

total # of surviving hares

# of surviving brown hares = frequency of surviving brown hares

total # of surviving hares

b. Determine the number of each color hares at the beginning of the next generation.

# of white hares beginning the next generation= Frequency of surviving white hares x 40
# of brown hares beginning the next generation= Frequency of surviving brown hares x 40

(e.g., 0.30 x 40 = 12 brown hares)

(e.g., 0.70 x 40 = 28 white hares)

4
 c. Record these values as the beginning numbers of brown and white hares in the next
generation of Table 3. To check you work, the beginning numbers of brown and white hares
should add together to equal 40. Ask the instructor to check your work before proceeding.
d. Spread the new generation’s beginning number of brown and white hares evenly on the dark
textured fabric and fluff the faux fur to help camouflage the hares.
7. Repeat steps 3-7 to determine which hares survived Generation Two and started Generation
Three. Make sure to record your data in Table 3. Continue this process until all data has been
collected through Generation Five.
If fixation occurs, then your lab group can stop the simulation at that generation.
Fixation occurs if either the brown or white hare number reaches zero and the surviving
color occurs in every member of the population. One allele has been lost from the
population and the other is fixed at 100%.
8. Record each lab group’s number of white moths and the beginning of each generation in Table 4.
These will be used to construct a graph in Activity 4.

Table 3. Lab group results from the simulation of population experiencing natural selection during the
summer (Activity 2).
Generation Number of Hares Number of Hares Numbers Surviving Frequency of
at the Beginning Captured Surviving Hares
of the
Generation
(Out of 40)
White Brown White Brown White Brown Total White Brown
One
Two
Three
Four
Five

Table 4. Class results from the simulation of natural selection (Activity 2).
Generation Number of White Hares at the Beginning of Each Generation
Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
One
Two
Three
Four
Five

5
Activity 3: Simulation of a Population Experiencing Natural Selection during the
Winter
You will simulate the predation of brown and white hares against a light background. The light
background will include a textured, white fabric that represents the snow-fallen ground during the winter
months in the taiga. Artic hares’ fur normally changes color from brown to white in the winter to
camouflage with the snow. Each lab group will determine if the population’s allelic frequencies of
brown and white hares are changing over time.

PROCEDURE:

1. Have the two predators look away while the other partners place 20 brown and 20 white-colored
hares in the plastic beaker, gently mix them, and randomly place them on the light textured
fabric where they are evenly spread out. Take a few minutes to fluff the fabric so the hares fall
in between the faux fur. This will represent your starting population (Generation One) of 40
hares that consists of 20 brown hares and 20 white hares.
2. Record the amount of brown and white hares in the beginning of Generation One (Table 5).
3. First round of predation:
a. One person will be the timer, two people will be predators, and the fourth person will be the
recorder.
b. The predators need to wear sunglasses and the instructor may choose to dim the lights before
predation begins.
c. When the timer says, “GO”, the predators will begin to feed for 10 seconds. The predators
will use their sight to capture one hare at a time and place it into a beaker. The predators
should capture prey as quickly as possible. After ten seconds, the timer says, “STOP”.
4. Record your lab group’s total “number of hares captured” (in the beakers) for each phenotype
(brown and white hares) for Generation One in Table 5.
5. Determine the “number surviving” brown and white hares using the formula below. Record
these values in Table 5.

# of surviving white hares = # of white hares at the beginning of the generation - # of total white hares
captured
# of surviving brown hares = # of brown hares at the beginning of the generation - # of total brown
hares captured

6. To build the population back up to 40, assuming the surviving hares produced offspring similar
to themselves:
a. Calculate the frequency of the surviving brown and white hares using the formula below.
Record the frequency of brown and white hares to two decimal places (e.g., you would round
0.456 to 0.46) in Table 5.

# of surviving white hares = frequency of surviving white hares

total # of surviving hares

# of surviving brown hares = frequency of surviving brown hares

total # of surviving hares

6
 b. Determine the number of each color moths at the beginning of the next generation.

# of white hares beginning the next generation= Frequency of surviving white hares x 40
# of brown hares beginning the next generation= Frequency of surviving brown hares x 40

(e.g., 0.30 x 40 = 12 brown hares)

(e.g., 0.70 x 40 = 28 white hares)

c. Record these values as the beginning numbers of brown and white hares in the next
generation of Table 5. To check you work, the beginning numbers of brown and white hares
should add together to equal 40. Ask your instructor to check your work before proceeding.
d. Spread the new generation’s beginning number of brown and white hares evenly on the dark
textured fabric and fluff the faux fur to help cover the hares.
7. Repeat steps 3-7 to determine which hares survived Generation Two and started Generation
Three. Make sure to record your data in Table 5. Continue this process until all data has been
collected through Generation Five.
8. If fixation occurs, then your lab group can stop the simulation at that generation. Fixation occurs
if either the brown or white hare number reaches zero and the surviving color occurs in every
member of the population. One allele has been lost from the population and the other is fixed at
100%.
9. Record each lab group’s number of white moths and the beginning of each generation in Table 6.
These will be used to construct a graph in Activity 4.

Table 5. Lab group results from the simulation of population experiencing natural selection during the
winter (Activity 3).
Generation Number of Hares Number of Hares Numbers Surviving Frequency of
at the Beginning Captured Surviving Hares
of the
Generation
(Out of 40)
White Brown White Brown White Brown Total White Brown
One
Two
Three
Four
Five

Table 6. Class results from the simulation of natural selection (Activity 3).
Generation Number of White Hares at the Beginning of Each Generation
Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
One
Two
Three
Four
Five

7
Activity 4: Interpretation of Class Results from the Simulations of Genetic Drift and
Natural Selection
Make sure your graphs for Figure 2, 3, and 4 all have the same x and y-axis scales. Label your x-axis
with “Time” (generations) and your y-axis “Number of White Hares” (beginning of generation sizes).
Create a key using different symbols (e.g., square, circle) or colors to represent each lab group.
Construct a graph that connects the data points with straight lines. Label each line G1, G2, etc. to
represent each lab group.

PROCEDURE A: Interpretation of Effect of Genetic Drift with Bottleneck Effect

1. To complete Figure 2, make a line graph of the class results (Table 2) from Activity 1.

Figure 2. Effects of genetic drift on the number of white hares over several generations.

8
Procedure B: Interpretation of Natural Selection during Summer Simulation
1. To compete Figure 3, make a line graph of the class results (Table 4) from Activity 2.

Figure 3. Effect of natural selection on the number of white hares during summer over several
generations.

9
Procedure C: Interpretation of Natural Selection Simulation
1. To compete Figure 4, make a line graph of the class results (Table 6) from Activity 3.

Figure 4. Effect of natural selection on the number of white hares during winter over several
generations.

Image Credits:
Cryptic Coloration Photos by OpenStax2e | [Link]
Graph Paper | [Link]

10
BI 101 Lab Worksheet: Microevolution Name ___________________________ Section _______

1. After reviewing the trends in Figure 2 (from Activity 1- Genetic Drift), did the frequency of
white hares consistently increase or decrease over several generations, or were the changes in the
frequency of white hares random from one generation to the next? Explain.

2. Did you see any of the alleles in the populations reach fixation? (Refer to Figure 2) If so, is
there a pattern to which specific allele reached fixation among groups? Explain.

3. If a larger number of individuals survived the catastrophe in Activity 1, would fixation be more
or less likely to occur? Why?

4. After reviewing the trends in Figure 3 from Activity 2-Natural Selection for your lab group, did
the frequency of white hares consistently increase or decrease over several generations, or were
the changes in the frequency of white hares random from one generation to the next?

5. For Activity 3-Simulation of a Population Experiencing Natural Selection during Winter Months,
how did the changes in frequency of white hares compare among lab groups? Were the results
from Figure 4 similar or did they vary among lab groups?

6. Compare the results from Activity 3 and 4 (Figure 3 and 4). Is there a selective advantage for
each fur color during the respective seasons? Why or why not?

11
BI 101 LAB 11
Predation and Competition
Prepared by Dr. Robin Whitekiller, Dept. of Biology, UNA
OBJECTIVES
After completing these laboratory activities, you should understand and be able to:
• Define population, ecology, predation, foraging success, camouflage, intraspecific competition, and
interspecific competition.
• Learn about the factors that affect foraging success: prey density, whether the prey are camouflaged,
whether other competitors are present, and the handling time of prey.
• Determine other variables that affect foraging success (e.g., mobility of prey).

Introduction
A population is a group of individuals of one species living in a particular area. Ecology is the study of
the interactions between individuals within a population and the environment. Predation occurs when
one organism preys on another. Foraging success is the number of prey collected per unit of time.
Foraging success will determine whether an individual will survive (e.g., over harsh winter months) and
whether an individual will be able to produce offspring. There are numerous factors that can affect an
individual’s foraging success including (but not limited to) the density of prey, whether the prey are
camouflaged (i.e., their color or color pattern allows them to blend into their surroundings), whether
there is competition for prey from others and handling time of the prey. For today’s activities, we will be
looking at carnivores or animals that eat other animals.

Activity 1: Foraging Success Based on Prey Density

We might hypothesize that individuals will spend more time foraging when prey density (the number of
prey in a given area) is lower. It should take longer to locate prey if there are fewer prey available.
PROCEDURE: Work in pairs for this activity. To test this hypothesis, we will set up a habitat
(container of rice) with prey (navy beans). The navy beans will be relatively camouflaged against the
rice for this activity. Each individual will forage twice while standing. The density of prey will be 15 the
first time and 30 the second time. Each individual foraging should not see where the prey are hidden in
the container; therefore, ask your partner to turn around while hiding the prey. Be sure prey are
randomly dispersed and hidden (covered by rice). Predator 1 will forage until 10 prey are captured using
one pair of tweezers to find (and capture) a prey item (a bean) and place it into a cup. Record the time in
Table One in seconds (s). The other individual will need a stopwatch to determine time spent foraging.
Once the individual has captured 10 prey, stop and record the time. Set up the container up with 30 prey.
The first predator will forage again until 10 prey are captured. Predator 2 will perform the same activity
with both prey densities. Record the values in Table One (on the next page) and calculate the averages
for the two individuals for both prey densities.

1
Table One: Time spent foraging based on density of the prey.

Predator(s) Prey density Time (sec)

1 15
1 30
2 15
2 30
1&2 15 Average:
1&2 30 Average:

Activity 2: Foraging Success when Prey are not Camouflaged

Many prey are camouflaged which provides some protection against predation. Other prey are more
visible. We might hypothesize that time spent foraging should decrease when prey are more visible.
PROCEDURE: Work in pairs. Be sure that the person foraging does not see where prey are hidden.
The tray (container with rice) will be set up with 15 prey randomly distributed and hidden; however, the
prey will be clearly visible once they are uncovered. We will work with red kidney beans this time. Each
predator will foraging while standing. Predator 1 will forage until he or she is able to capture 10 prey.
Record the time in Table Two. Reset the tray with prey randomly distributed and hidden. The second
predator will forage. Record the time. Calculate the average foraging time for both predators and record
the value in Table Two.

Table Two: Time spent foraging when prey are not camouflaged.

Predator Times (sec)

1
2
1&2 Average:

Activity 3: Foraging Success when Another Predator, Competing for the Same Prey,
is Present

So far, we have looked at how prey density and prey coloration may affect foraging success. We have to
remember that many individuals experience competition for prey with others within their own species
(intraspecific competition) and/or other species (interspecific competition). Some individuals will
even steal food from others. We might hypothesize that time spent foraging for prey will increase with
competition from another predator. PROCEDURE: To test this hypothesis, we will set up a habitat
(container of rice) with prey (navy beans). This time both individuals will forage at the same time (while
standing across from each other). The density of prey will be 15. The foraging individuals should not see
where the prey are hidden in the container; therefore, ask both predators to turn around while someone

2
from another table hides the prey. Each predator can attempt to interfere with the other’s ability to
capture prey, but do not injure each other or make a mess. An individual can steal a prey item IF it has
not made it into a cup. Whenever one student successfully collects 10 prey, stop the competition and
record the time for that individual in Table Three.
Table Three: Time spent foraging with a competitor present.

Predator # Time spent foraging when

a competitor is present
(sec)

Activity 4: Foraging Success when the Handling Time of Prey is Increased

Prey are not always easy to capture. Some prey are fast, while others have antipredator mechanisms.
For exam, porcupines have quills. They are sharp and able to pierce the skin of would-be predators.
Skunks can spray from anal scent glands. The chemicals in the spray irritate the eyes and nose of
predators. PROCEDURE: To test this hypothesis, we will set up a habitat (container of rice) with prey
(navy beans). The density of prey will be 15. The foraging individuals should not see where the prey are
hidden in the container; therefore, ask the predator to turn around while the prey are hidden. To simulate
increased handling time, the predator must capture the prey then alternate between touching the prey or
blunt end of the tweezers to his or her chin and back to the table five time before placing it in the cup
and attempting to capture another prey. Do NOT cut yourself or others with the tweezers. When the
predator has captured 10 prey, record the time in Table Four. Reset the tray with prey randomly
distributed and hidden. The second predator will forage. Both predators should stand to forage.
Calculate the average foraging time for both predators and record the value in Table Four.
Table Four: Time spent foraging when the handling time of prey increases.

Predator Times spent foraging

with increased
handling time (sec)
1
2
1&2 Average:

Activity 5: Collect Class Data, Calculate Averages and Construct a Bar Graph

When you finish with your activities, please share your group’s table values with the instructor. The
information will be entered in Table Five on the next page. Copy the data and calculate averages for
time spent foraging (seconds) for each of the variables: prey density 15, prey density 30, with prey that
are not camouflaged, with a competitor and with increased handling time. The prey density of 15 will
serve as our control. These prey were camouflaged, the density was 15, no competitor was present and
3
there was no increase in handling time. Construct a bar graph on page six with the five variables along
the X-axis and Time Spent Foraging (sec) along the Y-axis. You will need to label the X-axis using the
abbreviations provided (e.g., PD 15 for prey density 15). Use the class averages to answer the questions
in the Lab Worksheet.
Table Five: Average time spent foraging for each of the variables
Group Prey Prey Prey not Competitor Increased
density 15 density 30 camouflaged present handling time
(PD 15) (PD 30) (PNC) (CP) (IHT)
1
2
3
4
5
6
7
8
9
10
11
12
AVERAGES

4
BI 101 Lab Worksheet: Predation and Competition Name ____________________ Section ___

Activity 1: Foraging success based on prey density

1. Define predation.

2. What is one way a predator might be affected if its foraging success declines or is too low?

3. Based on average time spent foraging, do the results support our hypothesis? Did individuals spend
more time (on average) foraging when prey density was lower? Include data in your answer. Remember
to use class averages.

4. In the real world, prey are mobile. They are looking for food, too. If the prey had been mobile, would
the time spent foraging increase or decrease?

Activity 2: Foraging success when prey are not camouflaged

5. Based on the average time spent foraging, do the results support our hypothesis? Did individuals
spend less time foraging when the prey were more visible? Include data in your answer. Remember to
use class averages.

6. If prey are not camouflaged but brightly colored (e.g., coral snakes), how may they escape predation?
You may need to ask the instructor for assistance.

Activity 3: Foraging success when another predator, competing for the same prey, is present
7. Define the following types of competition:
A. intraspecific competition –

B. interspecific competition -

8. Based on the predator’s time spent foraging, do the results support our hypothesis? Did the time spent
foraging increase with competition from another predator? If not, explain why. Include data in your
answer. Remember to use class averages.

5
Activity 4: Foraging success when the handling time of prey is increased
9. Based on the predator’s time spent foraging, do the results support our hypothesis? Did the time spent
foraging increase with the increased handling time? Include data in your answer. Remember to use class
averages.

10. Most predators need to avoid becoming prey while foraging. Would the need to avoid predation
increase or decrease foraging success? Briefly explain.

AVERAGE TIME SPENT FORAGING BASED ON DENSITY OF PREY, FORAGING FOR PREY
THAT ARE NOT CAMOUFLAGED, FORAGING WITH A COMPETITOR AND FORAGING WITH
INCREASED HANDLING TIME

6
BI 101 LAB 12
Biodiversity and Ecological Interactions

Prepared by Dr. Emily Kasl, Dept. of Biology, UNA

OBJECTIVES
After completing these laboratory activities, you should understand / be able to:
• Define the concept of biodiversity and its application at macro and micro scales.
• Identify and explain the symbiotic partnership between termites and termite flagellates.
• Identify and explain the symbiotic partnership between Azolla and Anabaena.
• Use a quadrat sampling technique to investigate fundamental and realized niches of barnacles.

Introduction
Biodiversity is the variation in genes, species, and ecosystems that are found on Earth. The currently
observed biodiversity is a result of evolution, is an incredibly broad topic. In fact, an entire laboratory
(or course) could be devoted to only tackling that concept! The importance of recognizing the variety of
species and environments on Earth is two-fold.

First, appreciation of biodiversity is intrinsically valuable. We can have great appreciation for the
sound of bird calls, or the smell of flowers, or even an Alabama road trip from the Appalachian foothills
to the salty air of Gulf Shores. In the intrinsic sense, biodiversity is important because it exists as a part
of our natural world. Conserving species and maintaining the environment is thus our (human)
responsibility because it exists, regardless of its utility.

But utility is also important to consider. Biodiversity also has practical values that are directly
important to human values and services. For instance, photosynthetic organisms (algae, plants) produce
oxygen via photosynthesis which we need to breathe (conduct cellular respiration). Plants and animals
provide food. Medicines are often developed from the chemicals produced by bacteria, fungi, and plants.
Even our creative endeavors (such as tv and movies, art, and engineering) take inspiration from the
natural world. This lab exercise provides you the opportunity to investigate some of the diverse
structures and ecological interactions that are easily overlooked by casual observers.

An important aspect of biodiversity is interactions, both between organisms and between organisms and
their environments. These interactions are studied within the field of ecology. Symbiosis is a term that is
broadly used to describe the interaction between two organisms where at least one organism benefits.
These interactions can have positive, negative, or neutral effects on the organisms involved. For
example, mutualism describes a relationship between two organisms which results in both organisms
benefiting. On the other hand, if one organism benefits but the second is harmed, the relationship would
be considered parasitism. A third type of symbiosis, commensalism, describes one organism benefiting
from the interaction while the other is not otherwise affected.

Organisms also interact with each other in antagonistic ways, competing for resources such as food or
habitat. In this lab, you will have the opportunity to investigate the impact of both beneficial
(mutualism) and detrimental (competition) interactions.
1
Activity I: Mutualism – Termite Microbiome
Termites, which are well known for eating wood, are not capable of digesting the wood they ingest.
Specifically, they are not capable of digesting the tough cellulose that comprises this plant material.
Instead, the termites themselves will acquire a diverse gut microbiome of bacteria and protists that are
capable of breaking down the cellulose in wood.

Living as mutualistic symbionts in the gut of termites, termite flagellates, of the genus Trichonympha,
digest wood (specifically cellulose) by releasing enzymes into the termite gut. These critical components
of the termite microbiome must be obtained by licking the rectal end of another colony member or by
ingesting fecal matter of other colony members. This symbiotic relationship is an excellent example of
obligate mutualism, whereby neither partner can survive without the other. Without flagellates,
termites would be unable to obtain the necessary nutrition; the flagellates, on the other hand, are
incapable of living outside of the hospitable gut habitat of the termite. Related flagellates are also found
in wood-eating cockroaches and other insects.

PROCEDURE:
Create your own wet mount slide of the guts of a termite. To locate the protists on the microscope slide,
look for pear-shaped, “fuzzy” cells. This fuzzy appearance is due to the presence of multiple flagella
extending from their cell membrane. Observe the motion of the flagella as these organisms move
around.

1. Lay a paper towel flat on your desk.

On top of the paper towel place a
clean microscope slide.
2. In the center of your slide place one
drop of 0.3% NaCl solution (Figure
1). This saline solution is isosmotic
with the contents of the termite gut to
prevent damage to the flagellates.
3. With fine tweezers, center a living Figure 1. Wet mount slide preparation.
termite in the drop 0.3% NaCl on the
slide. Using the tweezers, gently pull
the termite apart at the abdomen. This will rupture the gut and release the creamy-brown gut
contents on to the slide. If necessary, apply some pressure to the abdomen to release more contents.
4. Remove the body of the termite and gently lower the cover slip on an angle until the droplet is fully
covered (Figure 1).
5. Place the slide on your compound scope. Focus the slide first with the scanning objective (4x)
before switching to the low-power objective (10x). Scan the slide for movement: termite flagellates
often appear as if they are vibrating due to the numerous hair-like flagella extending from their
cell membrane. Note: these flagellates may be spread throughout the NaCl solution or clustered
alongside tissue.
6. Once you have located your flagellates, you can switch to the high-power objective (40x) for closer
inspection. Draw your observations on the end of lab worksheet (pg. 7). In your drawing label the
flagella. Answer the questions based on your observations.

2
Activity 2: Mutualism – Anabaena and Azolla
Another interesting case of obligate symbiosis is that of the
fern Azolla and the cyanobacterium Anabaena. When
observing your ferns, notice they are “planted” in water rather
than soil. While most plants are dependent on the fertilization
of limiting nutrients (such as nitrogen) via soil, Azolla is able
to obtain nitrogen directly from the atmosphere and is thus not
restricted to soil habitats. The key to this ability is the close
association with Anabaena.

Anabaena is a type of filamentous cyanobacteria forming long

chains of vegetative cells (Figure 3). In low-nitrogen
conditions, a few of these cells will differentiate into
specialized nitrogen-fixing cells called heterocysts. These
cells are capable of converting atmospheric, inorganic
nitrogen into a form that is usable by Azolla. In turn, Azolla
provides an enclosed environment within which the Anabaena
can live.
Figure 2. Anabaena.
PROCEDURE:

Observe the Azolla provided on each lab bench. After your initial
observations, make a wet mount slide to observe the microscopic components of this association.

1. Obtain a dime-sized piece of Azolla and place on a clean microscope slide. Add 2-3 drops of
distilled water.
2. Use tweezers to “chop up” the Azolla – pinch and/or gently grind the plant material with the tips of
your tweezers until it is a well mashed pile.
3. Using a 10µL mini-pipette, transfer liquid taken from the Azolla slurry and place onto the center of
a second, clean microscope slide. Repeat this transfer 10 times. This will ensure you have adequate
material to observe.
4. Add a coverslip (Figure 1) and observe using a compound scope. You should be able to identify the
two different organisms (Azolla and Anabaena) present using shape and color differences.
Answer the questions on the end of lab worksheet (pg. 7) based on your observations.

Activity 3: Competition – Niche Modeling

Competition, as you have observed in previous labs, is another common interaction between organisms;
however, unlike symbiosis, neither organism ultimately benefits from the interaction. If given the
opportunity, a given species would expand to fill its ecological niche (or role) within its environment. In
the absence of limitations, such as competition, the organism can take advantage of its fundamental
niche, having access to all the space and resources needed to survive. When competition is present,
resources are limited and organisms are restricted to their realized niche, the part of a fundamental
niche that an organism actually occupies.

3
In this activity you will have a chance to further explore the findings of a classic competition experiment
and use a typical ecological assessment technique. Joseph H. Connell’s 1961 work focused on
competition between two barnacle species that commonly co-occur in Scotland’s rocky intertidal
coastline: the larger, long-lived Balanus and the smaller, more desiccation-resistant Chthamalus.
Barnacles are a type of marine crustacean that encrust on rocks and hard structures that extend below the
tide line. As adults, they cannot move from where they have settled, but they can close their shells to
protect themselves from predators and drying out (desiccation) as the water level ebbs and flows
throughout the day. Because of the tides, barnacles spend part of each day fully submerged, and part of
the day exposed to air. Competition for access to the lower tidal zone, which stays submerged longer,
can be fierce.

PROCEDURE:

Quadrat sampling is a method used in ecology to estimate population abundance, density, and
distribution. Because barnacles are sessile, they are an ideal organism to use with this sampling
technique. Our intertidal study sites have been divided into 16 quadrats along a transect (straight line)
that extends from high tide to low tide along the study site. For this experiment we will be estimating the
abundance of each species of barnacle at three Scottish beaches. Site 1, Argyll and Bute, has on
Chthamalus mussels. Site 2, Loch Ailort, has only Balanus mussels. Site 3, Musselburgh Beach, has
both Balanus and Chthamalus present.

1. Work in groups. Get one set of alphabet chips (A-D). These will be used to randomize the plots
sampled.
2. Find Site 1, Argyll and Bute. Starting at the high tide line (row 1), randomly remove one chip from
the container. Count all barnacles in the corresponding square, making a note of each species.
Record the total in Table 1. Return the chip to the container prior to sampling the next quadrat.
3. Continue with this procedure as you progress down each row towards the low tide mark. You
should have 4 quadrats sampled and recorded for each site.
4. Repeat steps 2-4 for Site 2 (Loch Ailort) and Site 3 (Musselburgh Beach).
5. To better visual the differences in distribution between the barnacle species, estimate the
population size within each tidal zone. To do this, add together the number of barnacles counted
within the tidal zone quadrants (e.g., high tidal zone contains rows 1 and 2) and multiple by 4.
Record your estimates in Table 1.

Optional: Quadrat sampling is a useful tool to more quickly assess organisms, however, it is still a
method of estimation and as such can be prone to either under-sampling or over-sampling. Assess the
accuracy of your population estimates using the steps below:

1. To estimate the total population size (of each barnacle species), add together the estimated totals
for both higher and lower tidal zones at each site.
2. Obtain the actual population counts from your laboratory instructor and add them to Table 1.
3. Use the following formula to calculate the percent error between your population estimate and the
actual population:
𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎 𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝 − 𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒 𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝
% 𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒 = × 100
𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎 𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝

4
Table 1. Barnacle Sampling Data
Site 1: Argyll and Bute Site 2: Loch Ailort Site 3: Musselburgh Beach
Quadrat
Balanus Chthamalus Balanus Chthamalus Balanus Chthamalus

Row 1

Row 2

Estimated
Total - Higher
Tidal Zone

Row 3

Row 4

Estimated
Total - Lower
Tidal Zone

Optional:
Total
Population
Estimate
Actual
Population
Count

Percent Error
of Estimate

Once you have completed the previous observations, answer the questions on the end of lab worksheet
(pg. 8).

Image Credits:
Wet Mount Preparation by Jeremy Sato | CC-BY-NC-SA [Link]
oer/blob/master/figures/microscopy/[Link]
Anabaena sphaerica by Elapied | CC-BY-SA [Link]

5
BI 101 Lab Worksheet Name Section
Lab 12: Biodiversity/Ecological Interactions

1. Drawing from your personal experiences (both inside and outside the classroom), provide an
example of an intrinsic value and practical value of biodiversity.

Intrinsic value:

Practical value:

Activity 1: Symbiosis - Termite Microbiome

2. In your own words, define mutualism:

3. Draw the termite flagellate and label the flagella:

4. The relationship between termites and termite

flagellates is best described as an
mutualism.

5. What does the termite provide in this relationship?

6. What does Trichonympha provide? Figure 3. Drawing of termite

flagellate (400x magnification)

Activity 2: Symbiosis – Anabaena and Azolla

6. What is the function of a heterocyst?

7. Draw your observation of the Azolla and

Anabaena. Label (and color) each organism:

8. What does the Azolla provide in this relationship?

9. What does Anabaena provide?

Figure 4. Drawing of Azolla and
Anabaena (400x magnification)

7
Activity 3: Competition – Niche Modeling

11. Use the estimated total barnacle density at higher vs. lower tidal zones data from Activity 4
(Table 1) to make three bar graphs. The y-axis has already been provided for you. On the x-axis,
place the independent variable (higher tidal zone, lower tidal zone). For Figure 7, choose colors
or cross-hatching to denote each barnacle species and label their respective bars.

110 -
Estimated number of barnacles

80 -

50 -

20 -

Figure 5. Number Figure 6. Number Figure 7. Number of Chthamalus and Balanus

of Chthamalus of Balanus barnacles observed at higher and lower tidal
barnacles observed barnacles observed zones in Musselburgh Beach (Site 3).
at higher and lower at higher and
tidal zones in Argyll lower tidal zones in
and Bute (S1). Loch Ailort (S2).

12. Consider Musselburgh Beach (Site 3), which species was more competitive in the lower tidal
zone?

13. In your own words, define the concept of fundamental niche vs. realized niche.

14. Of the three sites, which site shows the fundamental niche of Chthamalus?

15. Which site shows the realized niche of Chthamalus?