Laboratory Manual Part 1

(Labs 1 – 7)

BIOL 1106
Biology for Science Majors I
Laboratory Course

San Jacinto College, South Campus

Table of Contents
Lab Safety Rules for Biology Lab....................................................................................................3
Lab #1 Metric Measure..................................................................................................................4
Lab #2 pH and Buffers..................................................................................................................11
Lab #3 Molecular Models.............................................................................................................19
Lab #4 Chemical Composition of Cells.........................................................................................28
Lab #5 Microscopy....................................................................................................................... 38
Lab #6 Prokaryotic Cells and Eukaryotic Animal Cells..................................................................46
Lab #7 Eukaryotic Plant Cells....................................................................................................... 60
Review for Lab Practical #1..........................................................................................................69

–2–
 Laboratory Safety

Lab Safety Rules for Biology Lab

(student copy)

Keep this copy of the lab safety rules for yourself.

These rules are designed to minimize the possibility of injury as you work in the lab. These
rules apply at all times including exam days and during open lab. A student who fails to comply
with these rules will be asked to leave the lab.

1. Closed-toe shoes must be worn at all times.

2. Eating, drinking, chewing gum, all tobacco products, and the application of makeup
or lipstick are absolutely forbidden in the lab at all times. Do not place anything in
your mouth while in the lab. Keep your hands away from your mouth.
3. Cell phones are only allowed for taking photos and only at the discretion of the instructor.
4. Place all extra clothing, books, purses, backpacks, and other paraphernalia on the tables
at the sides of the lab or in the designated areas inside the lab bench. The only thing on
the lab bench should be your lab manual. Take reasonable precautions to protect your
belongings – for example, don’t leave wallets in plain sight.
5. Only students enrolled in the course are allowed in the lab. Children are NOT allowed in
the lab at any time.
6. Tables must be cleaned with 70% alcohol before and after use. Note that the alcohol in
these bottles contains contaminants such as methanol and is absolutely unsafe to
drink.
7. All reagents and equipment must be returned to their proper places.
8. Wash your hands well before leaving the lab.
9. Report all cuts or other injuries to the instructor immediately.
10. If you are or think you may become pregnant, consult with your doctor to determine
any additional precautions that he or she may suggest.

–3–
 Lab #1 – Metric Measure

Lab #1 Metric Measure

Introduction

Throughout the world the metric system is an integral tool, from science to commerce.
Although the United States does not have the metric system as well integrated into society as
other countries in the world, it does play an important role. Based on the powers of 10 and
decimal notation, it allows for seamless conversions between large and small measures. For
the following measures the metric unit is in parenthesis: length (meter), weight (gram), volume
(liter), and temperature (Celsius).

Table 1 – Metric Prefixes

Length Prefix Abbreviation
Gigameter giga Gm 1000000000.= 109
Megameter mega Mm 1000000. = 106
kilometer kilo km 1000. = 103
meter m 1.0
decimeter deci dm 0.1 = 10-1
centimeter centi cm 0.01 = 10-2
millimeter milli mm 0.001 = 10-3
micrometer micro μm 0.000001 = 10-6
nanometer nano nm 0.000000001 = 10-9

These prefixes are the same for every metric unit. For example, 1 kilometer is 1000 meters
(when measuring length) and 1 kilogram is 1000 grams (when measuring weight). Likewise, 1
milliliter is 0.001 liters (when measuring volume) and 1 milligram is 0.001 kilogram.

Pre-Lab Questions

1. What prefix denotes 0.001 meter?

2. Name the metric unit of length. _
3. Name the metric unit of weight.
4. Name the metric unit of volume.
5. Name the metric unit for temperature. _
6. What is the abbreviation for nanometer? _
7. What prefix denotes 1000? _
8. What prefix denotes 0.1? _

–4–
 Lab #1 – Metric Measure

Purpose of the Lab

This lab will familiarize you with the basic units of the metric system and how to convert
between them. You will also learn how to measure length, weight, volume, and temperature
with an acceptable level of scientific accuracy.

Materials

Metric ruler
Meter stick
Bone
Wooden paddles
Test tube
Thermometer (Celsius)
Triple-beam balance
Plexiglass block
Fossil
Warm-water bath
Bucket with ice
Graduated cylinder
Plastic pipettes
Pipette Pump
Water

Lab Procedures and Data

Length
Measure the length of this line: cm

Convert this measurement to mm and nm.

Measuring irregularly shaped three-dimensional objects can be tricky. To make an accurate

measurement, we will use a meter stick and two wooden paddles.

1. First, place the meter stick on a flat surface.

2. Sandwich the object you are measuring between the two paddles, making sure the
paddles are at a right angle to the object you are measuring (or as close as
possible).
3. Place the object and paddles along the meter stick and note the location of the inner
edges of the paddles.
4. Subtract to find the length of the object.

–5–
 Lab #1 – Metric Measure

For example, if you are measuring a bone and one paddle is at 13 cm and the other is at 41 cm,
the bone is 41 cm – 13 cm = 28 cm.

The set-up for measuring irregularly shaped objects

How long is the bone in the picture above? cm = mm

Measure both bones at your lab bench in cm using the meter stick and wooden paddles – then
convert to mm.
Bone #1: cm = mm
Bone #2: cm = mm

–6–
 Lab #1 – Metric Measure

Weight
To measure weight, we use a triple beam balance. To use the triple beam balance:

1. IMPORTANT: Before using your balance, make sure it is aligned

properly. Make sure there is nothing on the balance, then look
at the right side. The arrow should be aligned with a small line. If
the two are not aligned, ask your instructor for assistance.
2. Carefully place the object you are going to weigh on the round
metal platform on the left side of the balance.
3. ALWAYS begin with the largest weight! Move the weight to the
left one-notch at a time (you should hear a small click or feel the weight drop into place
when it is in the correct position).
4. Keep moving the weight to the left until the arrow drops below the mark on the
right side of the balance (meaning it is heavier than the object).
5. Slide the weight you just moved BACKWARDS (to the right) ONE NOTCH until you
are again lighter than the object.
6. Repeat steps 4 and 5 with the smaller weight, then repeat again with the smallest
weight of all. This smallest weight does not have distinct notches – it just slides along
the beam. Slide it back and forth until the arrow is exactly aligned with the line on the
right side of the balance.

A correctly used triple beam balance.

What is the weight of the object in this picture? g= mg

Using the triple-beam balance weigh the fossil and plexiglass block, then convert the weight
from grams to milligrams.

Weight of fossil g= mg

Weight of plexiglass block g= mg

–7–
 Lab #1 – Metric Measure

Volume
Large volumes of liquids are most accurately measured in a graduated cylinder, which can
come in various sizes. Put 20 mL of water in your graduated cylinder and examine it closely.

You should have a curve at the surface of the water. Because of cohesion, the water clings to
the inner walls of the graduated cylinder, resulting in what we call a meniscus. IMPORTANT:
To measure the water volume ACCURATELY, you should always look at the surface of the water
at eye level (put your eye parallel to the surface of the water) and measure from the bottom of
the meniscus.

How much water is in the graduated cylinder on the right? mL = L

Graduated cylinders can also be used to measure the volume of solid objects. The volume of an
object is equal to the volume of water displaced when the object is submerged.

Use your graduated cylinder to determine the volume of the fossil.

ml = L

With the tools that you have on the table, how would you find the maximum volume of water
the test tube can hold?

Check your experimental plan above with your instructor, then carry it out. What is the
maximum volume (in mL) of water the test tube can hold? mL

Reusable pipettes are very common lab equipment, and you will be using them for the rest of
the semester. A pipette pump is used in combination with these pipettes to measure and
transfer liquid. Each pump has a thumb wheel – when you use your thumb to rotate the wheel,
this causes fluid to be taken up or released from the pipette, depending on the direction of
movement.

–8–
 Lab #1 – Metric Measure

1. Attach the pipette pump to the 10 mL pipette provided.

2. Use the thumb wheel to draw up a total of 10 ml of water into the pipette.
3. Now add 5 mL of water to the graduated cylinder. Notice that if you remove your
thumb from the wheel, the solution may leak out of the pipette!
4. Read the graduated cylinder to test your accuracy.

Now transfer 2.5 mL of water from the pipette into to the test tube. How much water is left in
the pipette?

What total volume can the smaller plastic pipette hold?

Use the smaller plastic pipette to add 0.5 mL of water to the test tube. Could you have
measured that accurately with the graduated cylinder? Explain why or why not.

Temperature
Temperature can be measured in either Celsius or Fahrenheit. For example, the average
human body temperature is both 98.6oF or 37oC. Most scientific labs use the Celsius scale.

What is the temperature? (make sure to use the proper unit!)

–9–
 Lab #1 – Metric Measure

o o
Freezing point of water is C and F
o o
Boiling point of water is C and F

Read the thermometer in the following locations and record the

temperature (make sure to use the correct unit!)

Lab table

Ice bucket

Warm water bath

Read the thermometer after you have held the bulb of

the thermometer in your hand for 2 minutes

Post-Lab Cleanup
Dump all used water out in the sink in the middle of the lab table or the sinks on the sides of
the lab. Make sure all the equipment is clean and in its proper place. Place test tubes upside
down in the rack at your desk so they can drain. Ethanol the desks and push in your chairs.

Post-Lab Review Questions 1.

Conversion Problems:

1 cm = mm 1 mm = µm 1 µm = nm 1mm = cm 1 µm = mm 1 nm = µm
1 km = m 23 cm = mm 3 nm = µm
185 mm = cm 6 µm = nm 0.7 mm = µm
500 mg = g 0.02 g = mg 10 mg = g
1 mL = L 75 mL = L 10 L = mL

1. Which part of the meniscus is used to read the volume of water in a graduated cylinder?

2. Do metric thermometers read in Celsius or Fahrenheit? _

3. What is the freezing point of water in Celsius? __
4. What is the freezing point of water in Fahrenheit? __________
5. What is the boiling point of water in Celsius? ___
6. What is the boiling point of water in Fahrenheit?

References

Hoobler, Cynthia, Karen Duston, Adam Eiler, Jennie Plunkett, Kirsten Raines, and Mary
Wisgirda. (2007). Metric Measure. General Biology I and II, 1-6.

– 10 –
 Lab #2 – pH and Buffers

Lab #2 pH and Buffers

Introduction

All of you interact with acids and bases daily. Remember the taste of lemon juice on your
tongue or using bleach when doing laundry? How acidic or basic these and other substances
can be determined using the pH scale. This scale is numerical, logarithmic, and based on the
concentration of H+ (hydrogen ions). The scale ranges from 0-14, with 7.1 and above basic, 7
neutral, and 6.9 and lower acidic.

You may have experienced discomfort from excess acid in your stomach and taken an antacid
for relief. Antacids have buffering capabilities. A buffer is a substance that resists changes in
pH. Our blood stream has buffers to prevent swings out of a very narrow pH range (around
7.4). A bloodstream pH out of this range can have serious to lethal consequences.

Pre-Lab Questions

1. Is a pH of 11 acidic or basic?
2. Is a pH of 2 acidic or basic?
3. Does our bloodstream need buffers? Why or why not?

4. The pH scale is based on what?

5. What is the purpose of this lab?

Materials
pH paper
Common Household Liquids to Test
Inorganic buffer
Organic buffer
Several different antacids
Pipette
Wax pencil
Pipette
Pipette pump
Stirring rod
Test tubes
Test tube rack
Beakers
Bromocresol purple (pH indicator)

Lab Procedures and Data

– 11 –
 Lab #2 – pH and Buffers
The pH paper we will use in this lab can detect both acids and bases. To read the pH:
1. Tear off a small (~ ½ inch) piece of pH paper.
2. Add one drop of liquid to the paper.
3. As quickly as possible, compare the paper to the scale on the outside of the
package (the color will change over time, resulting in a less accurate reading).
4. Decide on the best pH reading by comparing the colors. Sometimes, you may record a
pH that is in between two different numbers (e.g. 4.5)

Left strip – acidic pH (<7)

Middle strip – neutral pH
(7) Right strip – basic pH
(>7)

Determine the pH of various substances using pH paper.

Examine the various substances on your lab bench. For each, make a hypothesis – do you think it is an
acid, a base, or neutral? Test your hypothesis using the provided pH paper. Also, record the pH of
Hydrochloric Acid (HCl) and Sodium Hydroxide (NaOH, a strong base).

– 12 –
 Lab #2 – pH and Buffers

Aside from the HCl, what was the strongest acid tested?
Aside from the NaOH, what was the strongest base tested?
Did the pH of any of the substances surprise you? Explain.

Why would tap water and rainwater not have the same pH?

Cells and pH

This next section will demonstrate how buffers in living cells help to regulate pH. To perform
the experiment:

1. Label the beakers 1, 2, and 3.

2. Add 6.5 mL of water to beaker 1, 6.5 mL of inorganic buffer to beaker 2, and 6.5 mL
of organic buffer to beaker 3. The buffers can be found at the front of the room.

3. Use the stirring rod to add one drop from each beaker to pieces of pH paper, and
determine the pH of the solutions in all three test tubes. Make sure you use a
different stirring rod for each beaker to avoid contamination!

4. After adding 5 drops of HCL to all three beakers, determine the pH of each solution
by dipping the stir rod into the solution then touching it to a small piece of pH paper.

5. Continue adding drops of HCl to beakers 2 and 3 in increments of 5 drops, determining

the pH after each addition of HCl, and recording the pH in the chart.

6. You should continue adding drops of HCl until you have added 50 drops to each beaker.
You should then graph your results.

– 13 –
 Lab #2 – pH and Buffers

Comparison of Organic and Inorganic Buffers

pH in Beaker 2 pH in Beaker 3
Number of Drops
( buffer) ( buffer)
0 (Initial pH from previous
table)
5 (Ending pH from previous
table)

10

15

20

25

30

35

40

45

50

– 14 –
 Lab #2 – pH and Buffers

What was the effect of having a buffer (either organic or inorganic) in the solution?

Which buffer became acidic faster, the organic or the inorganic?

Which was the better buffer, the organic or the inorganic? How do you know?

– 15 –
 Lab #2 – pH and Buffers
Finding the Strongest Buffer

Antacids help to reduce stomach acidity. They are usually weak bases that can also act as
buffers. This helps to prevent damage to the stomach. In this portion of the lab we will test
several different antacids to determine which is the best buffer.

Which antacid do you think will be the best buffer?

To test the buffers:

1. Take enough test tubes to test each buffer found at the back of the room. Number each
test tube (1, 2, 3, etc).

2. Add 4.5 mL of a different antacid to each test tube. Stir each antacid well before
pipetting and rinse the pipette with distilled water between each antacid. Make sure to
record which antacid is in each test tube.

3. Add 4 drops of Bromocresol purple to each test tube. Bromocresol purple is a pH

indicator. A purple color indicates the solution is basic, while yellow shows that the
solution is acidic.
What color is the solution now?
Does this mean the solution is acid or basic?

4. Add HCl to each antacid, one drop at a time, until the solution turns yellow. For each
antacid record the number of drops it takes to make the solution change color.

– 16 –
 Lab #2 – pH and Buffers

What was the final color of each test tube?

Does this mean the solution is acidic or basic?
Which antacid was the strongest buffer?
Explain your reasoning.

Post-Lab Cleanup

Clean all test tubes with soap and water either in the sink in your lab table or the sinks along
the edges of the room. MAKE SURE ALL WAX PENCIL MARKS ARE REMOVED. Place test tubes
upside down in rack to dry. Ethanol the desks and push in the chairs.

Post-Lab Review Questions

1. If the pH paper turns a bright red, does that indicate that the substance is an acid or a base?
________________________________________________________________________________
________________________________________________________________________________
2. If you are presented with two tubes, one tube with a buffered solution + acid and one tube with
water + acid, how will you know which tube has the buffer and which tube does not have the
buffer? _________________________________________________________________________
________________________________________________________________________________
3. Name the pH indicator used in the antacid experiments. __________________________________
4. What color is the antacid solution when the pH indicator is first added? _____________________
5. What color is the antacid solution/pH indicator when the antacid is no longer effective?
______________
6. You are presented with two test tubes; one has a yellow substance, and one has a purple
substance. What is the color of the tube with the lowest pH number? ______________________
7. You are presented with two test tubes; one has a yellow substance, and one has a purple
substance. What is the color of the tube with the higher pH number? ______________________
– 17 –
 Lab #2 – pH and Buffers
8. You are presented with two tubes; one has a yellow substance, and one has a purple substance.
Which tube indicates the most effective antacid? Explain your reasoning. ____________________
________________________________________________________________________________
9. Which antacid tested in this lab was the most effective? __________________________________
10. If a solution has a pH of 11, is that solution an acid or a base? ______________________________

References

Brooker, Robert, Eric Widmaier, Linda Graham, and Peter Stiling. (2014). The Chemical Basis of
Life I: Atoms, Molecules, and Water. Biology 3rd Edition, 21-41.

Hoobler, Cynthia, Karen Duston, Adam Eiler, Jennie Plunkett, Kirsten Raines, and Mary
Wisgirda. (2007). pH and Buffers. General Biology I and II, 45-48.

– 18 –
 Lab #3 – Molecular Models

Lab #3 Molecular Models

Introduction

Atoms are the smallest unit of matter. When two or more atoms bond, a molecule is formed.
The result of the bonding between atoms of two or more different elements is a compound. Of
all the elements, carbon is of special importance because it enables the diversity we see in life.
Compounds which contain carbon are organic compounds, while those compounds that do not
contain carbon are referred to as inorganic compounds. The different properties which organic
compounds have are determined by the functional groups which they contain. The following
are some examples of functional groups:

 NH2 (amino group),

H
N
H

COOH (carboxyl group),
O

C
O H

OH (hydroxyl group)
O H

“R” group (a variety of different groups of atoms which make one amino acid
different from another). The R group is much like the “x” in an algebraic equation.

H H These are only

a few of the
H C H C S H many possible
R groups
H H
A diagrammatic representation of how a molecule is constructed is its structural formula.
When both the number and type of atoms in a particular molecule are given, it is referred to as
an empirical formula.

H 2O
Empirical formula Structural formula
of water of water

– 19 –
 Lab #3 – Molecular Models

Pre-Lab Questions

1. Define an organic compound.

2. What is a molecule?
3. What is a structural formula?
4. What is the smallest unit of matter?
5. Define an empirical formula.

6. Are you permitted to wear flip-flops in the lab?

7. Are you permitted to chew gum or eat in the lab?

Purpose of the Lab

This lab will familiarize you with the ideas of atoms, molecules, and double and single bonds.
You will also learn to recognize several important functional groups and organic and inorganic
molecules.

Materials

Molecular Model Kit

Lab Procedures and Data

Examine the Molecular Model Kit:

The following atoms are represented in your molecular model kit:

Carbon (C) – can bond up to four different atoms

Nitrogen (N) – can bond up to three different atoms

Oxygen (O) – can bond up to two different atoms

Hydrogen (H) – can only bond to one other atom

Using this information, what atoms do the following colors represent?

Black Red Yellow Blue

Long sticks, short sticks, and the springs are used to form covalent bonds between the atoms.
Use the following rules to make covalent bonds between atoms:

When creating a single covalent bond, use a short stick.

When creating a carbon to carbon single covalent bond, use a long stick.

When creating a double bond, use two springs.

– 20 –
 Lab #3 – Molecular Models

Building Molecules

Using your kit, construct and name the following functional groups:

--COOH

Name:

--NH2

Name:

--OH

Name:

Construct models of the following molecules:

Water (perhaps the most important compound on Earth)

Is this molecule organic or inorganic?

What is its empirical formula?

Methane (the main component of natural gas, sometimes produced as a byproduct by

microorganisms)

H C H

H
Is this molecule organic or inorganic?
What is its empirical formula?

– 21 –
 Lab #3 – Molecular Models

Ethane (a colorless, odorless gas produced as a byproduct of petrochemical refining. It is

a common contaminant of moonshine and can make you blind.)

H H

H C C H

H H
Is this molecule organic or inorganic?
What is its empirical formula?

Ethyl alcohol (Also called ethanol, it can be used as a fuel source. It is also the alcohol found in
adult beverages).

H H

H C C H

H OH
Is this molecule organic or inorganic?
What is its empirical formula?

Ribose (Ribose is a 5-Carbon sugar molecule found in RNA strands. RNA stands for
RiboNucleic Acid)

H H OH H O
H C C C C C

OH OH H OH H
What is the empirical formula?
Why is this molecule called a “pentose sugar?” (Hint: What does “pent-” mean?)

– 22 –
 Lab #3 – Molecular Models

Deoxyribose (This is the sugar found in DNA strands. DNA stands for DeoxyriboNucleic Acid).

H H OH H O
H C C C C C

OH OH H H H
What is this molecule’s empirical formula?
What is the difference between this molecule and ribose?

What does “deoxy-“ mean?

Fructose (The monosaccharide (simple sugar) is found in many plants (for example,
high fructose corn syrup comes from corn). It is also found in honey)
OH H OH H O H

H C C C C C C OH

H OH H OH H

Is this molecule organic or inorganic?

What is its empirical formula?

Glucose (This monosaccharide is the main energy source for most animals)

OH H H OH H O
H C C C C C C

H OH OH H OH H
What is this molecule’s empirical formula?
Do glucose and fructose have the same empirical formula?
Do they have the same structural formula?

– 23 –
 Lab #3 – Molecular Models

Glucose (ring) (Glucose can be stored in the body in its “ring” form. Many rings can be linked
together to form long, very compact chains. These chains of glucose are unbranched in
plants (starch) and branched in animals (glycogen)).

What is this molecule’s empirical formula?

Do linear glucose and ring glucose have the same empirical formula?
Do they have the same structural formula?
Why do you think glucose is called a hexose sugar?

Fructose can also form a ring structure:

– 24 –
 Lab #3 – Molecular Models

Sucrose (disaccharide) In their ring structures, glucose and fructose (two monosaccharides) can
bind together to form a disaccharide called sucrose. Sucrose is also called table sugar and
usually comes from sugar cane or sugar beets.

What does the prefix “di-“ mean in “disaccharide”?

What do you think the suffix “-ose” means?
What atom is removed from the fructose to form sucrose? (red boxes)
What functional group is removed from the glucose? (blue boxes)
What molecule is formed when these atoms combine?

Amino Acids
Glycine and alanine are amino acids. There are 20 amino acids differentiated by various “R”
groups. Note that each amino acid contains carboxyl and amino groups.

– 25 –
 Lab #3 – Molecular Models
What two functional groups are present on both of these molecules?

The R group is the only functional group that is different on different amino acids.
What R group is present on glycine?
What R group is present on alanine?

Amino acids link together to form peptides, or proteins. When two amino acids are joined
together it forms a dipeptide, while three form a tripeptide, and many form a polypeptide.
Amino acids can only be added to the carboxyl end (--COOH) of a growing peptide.

What atoms were removed from glycine when you made the dipeptide?
What atom was removed from alanine when you made the dipeptide?
What molecule can the atoms you removed combine together to make?

Working together with another group in your class, join together three amino acids to form a tripeptide.
Sample tripeptides include:
Glycine-Glycine-Glycine
Glycine-Alanine-Alanine
Alanine-Alanine-Glycine
(Each amino acid in the tripeptide could be either alanine or glycine).

How many water molecules were also made when you made the tripeptide? How many different
tripeptides can be formed from these two amino acids?

List all the possible tripeptides below:

Post-Lab Cleanup

Count all balls, sticks, and springs to make sure you have the correct amount. Bundle sticks and springs
together with rubber bands. Place all balls, sticks, and springs back into the box. Ethanol the desks and
push in the chairs.

Post Lab Review Questions

– 26 –
 Lab #3 – Molecular Models

1. What atom does the blue ball represent? __________________

2. What atom does the red ball represent? __________________
3. What atom does the black ball represent? _________________
4. What atom does the yellow ball represent? ________________
5. What do the springs represent? _____________________________________________________
6. Is water an organic or inorganic molecule? ____________________________________________
7. Is ethyl alcohol an organic or inorganic molecule? ______________________________________
8. How can you tell the difference between ribose and deoxyribose? __________________________
________________________________________________________________________________
9. Name the amino acids we made in the lab. Give the empirical formula for the R group found on
each of them. ____________________________________________________________________
10. Name this functional group: -NH2 ___________________________
11. Name this functional group: -COOH _________________________
12. What two molecules are produced when the two monosaccharides fructose and glucose are
combined? ______________________________________________________________________
13. What two molecules are produced when two amino acids are combined? ___________________

References

Brooker, Robert, Eric Widmaier, Linda Graham, and Peter Stiling. (2014). The Chemical Basis of
Life I: Atoms, Molecules, and Water. Biology 3rd Edition, 21-41.

Hoobler, Cynthia, Karen Duston, Adam Eiler, Jennie Plunkett, Kirsten Raines, and Mary
Wisgirda. (2007). Molecular Models. General Biology I and II, 7-11.

– 27 –
 Lab #4 – Chemical Composition of Cells

Lab #4 Chemical Composition of Cells

Introduction
In living organisms and in common substances that we use daily, the large biological molecules
– proteins, lipids, nucleic acids, and carbohydrates – are present. By utilizing different chemical
reagents, and for one experiment a simple brown paper towel test, you will be able to
determine (with the exception of nucleic acids), if these large macromolecules are present in a
variety of substances. In all the experimental procedures water will be the control. A control is
a substance included in the experiment that we already know does not contain the
macromolecule we are testing for. This is important to ensure the accuracy of the experiment.

A different reagent will be used for each molecule we are testing for. This reagent will have
certain chemical properties, and there will be a visible change in the reagent when it interacts
with the molecule we are testing for. A positive test for a molecule will be indicated by a color
change as compared with the control (meaning the substance we are testing for is present). If
the reagent is the same color as the control, the result is a negative test, and we know that the
substance we are testing for is not present.

Left test tube (water): the negative control

Right tube (starch): color change = a positive test result

– 28 –
 Lab #4 – Chemical Composition of Cells

Pre-Lab Questions

1. What substance is used as the control?

2. What reagent is used to test for protein?
3. What reagent is used to test for starch?
4. What reagent is used to test for sugar?
5. How do you test for lipids?
6. What is the purpose of today’s lab?

7. What safety precautions are needed for today?

Materials

Wax pencil
Test tubes
Test tube rack
Test tube holder
10 mL pipette
Pipette pump
1.5 mL pipette
Biuret reagent
Iodine
Benedict’s reagent
Brown paper towel
Albumin
Pepsin
Starch
Glucose
Onion juice
Potato juice
Distilled Water
Unknown
Vegetable oil
Boiling water bath

– 29 –
 Lab #4 – Chemical Composition of Cells

Lab Procedures and Data

Protein Test: Biuret’s reagent reacts with the peptide bonds found in proteins. If a test is
positive for protein the substance will turn purple. A negative test will remain light blue.

Test reagent: Biuret

Test procedure:
The following solutions will be tested to determine if they contain or do not contain protein:
albumin (egg whites) (test tube 1), pepsin (an enzyme) (test tube 2), starch (test tube 3),
unknown (test tube 4), and distilled water (test tube 5).

Hypothesis: Of the above solutions, which do you think will contain proteins?

1) Label five test tubes with the numbers 1 through 5.

2) Add 3 mL of the correct solution to each test tube. Rinse the pipette with distilled
water between each use. Record the initial appearance of the solution in the table
below.
3) Use the small pipette to add 10 drops of biuret reagent to each test tube, and
then swirl to mix.
4) Record the final appearance of each test tube. In the results column, indicate
whether each solution tests positive or negative for protein.

Results of the Biuret Test for Protein

Tube Initial appearance of Final appearance of

Solution Results
Number solution solution
1

– 30 –
 Lab #4 – Chemical Composition of Cells

What was the control for this experiment? Did the control perform as expected?

What is pepsin? Explain why it makes sense that a pepsin solution contains proteins.

Carbohydrates come in two main forms – monosaccharides and polysaccharides.

Monosaccharides are single-sugar units (e.g. glucose and fructose):

glucose fructose .

Polysaccharides are formed by linking together many monosaccharides. There are several
different types of polysaccharides. Starch is an unbranched polysaccharide used for energy
storage in plants, while glycogen is a heavily branched polysaccharide used for energy storage
in animals. Cellulose is polysaccharide used for structural support in plants – in makes up the
cell wall that surrounds most plant cells. Humans cannot digest cellulose; it passes straight
through our digestive system as dietary fiber.

– 31 –
 Lab #4 – Chemical Composition of Cells

Starch Test: If a test is positive for starch the solution will turn blue-black. The color of the
negative test will be yellow brown.

Test reagent: iodine

Test procedure:
The following solutions will be tested to determine if they contain or do not contain starch:
glucose (test tube 1), starch suspension (test tube 2), unknown (test tube 3), distilled water
(test tube 4), onion juice (test tube 5), and potato juice (test tube 6).

Hypothesis: Of the above solutions, which do you think will contain starch?

1) Label six test tubes with the numbers 1 through 6.

2) Add 1.5 mL of the correct solution to tubes 1-6. Make sure to stir the solutions well
before pipetting them and rinse the pipette after each use.
3) Record the initial appearance of each solution in the table below.
4) At this point add 10 drops of iodine to each test tube. Swirl each test tube to mix.
5) Record the color change in each test tube. In the results column of the
table, indicate whether each solution tests positive or negative for starch.

Results of the Iodine Test for Starch

Tube Initial appearance of Final appearance of

Solution Results
Number solution solution

– 32 –
 Lab #4 – Chemical Composition of Cells

What was the control for this experiment? Did the control give the expected result?

Does glucose give a positive result for starch? Explain why or why not.

Sugar (monosaccharides)Test: This test is only for monosaccharides, not polysaccharides like
starch. If a test is positive for sugar it will turn green, yellow, yellow-orange, orange, or orange-
red. The color changes indicated (beginning with green) are in order of increasing
concentrations of sugar.

Test reagent: Benedict’s

Test Procedure: The following solutions will be tested to determine if they contain or do not
contain sugar: glucose solution (test tube 1), starch suspension (test tube 2), unknown (test
tube 3), distilled water (test tube 4), onion juice (test tube 5), and potato juice (test tube 6).

Hypothesis: Of the above solutions, which do you think will contain monosaccharides?

1) Label six test LARGE test tubes with the numbers 1 through 6.
2) Fill test tubes 1-6 with 1.5 mL of their respective solutions. Rinse the pipette
between solutions.
3) Then add 2 mL of Benedict’s reagent to all six test tubes. Record the initial
appearance of the solution in each test tube.
4) Place all test tubes in a boiling water bath at the same time.
5) After 5 minutes, carefully use the test tube clamps to remove the test tubes
from the boiling water.
6) Let the test tubes sit for 1-2 minutes to cool, then gently shake the test tube to mix
and suspend any precipitate (solid particles at the bottom of the test tube).
7) Record the color of the solution in each test tube. In the results column, indicate
whether each solution tests positive or negative for sugar.

– 33 –
 Lab #4 – Chemical Composition of Cells

Results of the Benedict’s Test for Monosaccharides

Initial Appearance of Final appearance
Tube
Solution appearance solution after of solution after Results
Number
of solution adding Benedict’s heating
1

What was the control for this experiment? Did you see the expected result?

What result did you see for the glucose solution? Explain this result.

What result did you see for the starch suspension? Explain this result.

Compare the results of this test with those from the starch test. Which plant – onions or
potatoes – store most of their carbohydrates as monosaccharides? How do you know?

Which plant – onions or potatoes – store most of their carbohydrates as polysaccharides?

Explain your answer.

– 34 –
 Lab #4 – Chemical Composition of Cells

Lipid Test: On brown paper a positive test for lipids shows an oily residue and no absorption.
By contrast a negative test will be absorbed by the paper and eventually evaporate.

The following solutions will be tested to determine if they contain or do not contain lipids:
vegetable oil, glucose solution, potato juice, pepsin, water and the unknown.

Hypothesis: Of the above solutions, which do you think will contain lipids?

Test Procedure:

1) Draw six circles on a large piece of paper towel (2 layers recommended). Label
these circles “oil,” “water,” “glucose solution,” “potato juice,” “pepsin,” and
“unknown.”
2) Place one drop of each liquid in the correct circle on the paper and observe
what happens.
3) Record your results. Indicate whether each substance tests positive or negative
for lipids.

Results of the Brown Paper Test for Lipids

Final appearance of
Circle Initial appearance of
Solution paper towel after 5 Results
Number liquid on paper town
minutes

– 35 –
 Lab #4 – Chemical Composition of Cells

What was the control for this experiment? Did it yield the expected result?

What would be another way to test if a solution contains lipids (HINT: are lipids hydrophobic or
hydrophilic)?

Summary of Results on the Unknown Solution

Test Observations Conclusions

Biuret/Protein

Iodine/Starch

Benedict/Sugars

Brown Paper/Lipid

Unknown solution:

Post-Lab Cleanup

Clean all test tubes with soap and water either in the sink in your lab table or the sinks along
the edges of the room. MAKE SURE ALL WAX PENCIL MARKS ARE REMOVED. Place test tubes
upside down in rack to dry. Ethanol the desks and push in the chairs.

Post-Lab Review Questions

1. What reagent is used to test for proteins?

___________________________________
2. What color indicates the presence of protein? _______________________________
3. Is this color a positive or negative test for protein? ___________________________
4. What color indicates the absence of protein? ________________________________
5. Is this color a positive or negative test result for protein?
_______________________
6. What reagent is used to test for starch?
_____________________________________
7. What color indicates the presence of starch? ________________________________
8. Is this color a positive or negative test result for starch? _______________________

– 36 –
 Lab #4 – Chemical Composition of Cells
9. What color indicates the absence of starch?
_________________________________
10. Is this color a positive or negative test result for starch?
________________________
11. What reagent is used to test for monosaccharides (sugars)?
_____________________
12. What color(s) indicates the presence of monosaccharides? _____________________
13. Is this color a positive or negative test result for monosaccharides?
_______________
14. What color indicates the absence of monosaccharides? ________________________
15. Is this color a positive or negative test result for monosaccharides?
_______________
16. Describe the appearance of the paper towel when a lipid is present.
_______________
_____________________________________________________________________
17. Is this a positive or negative result? ________________________________________
18. Describe the paper towel when a lipid is not present.
__________________________
19. Is this a positive or negative result? ________________________________________

References

Brooker, Robert, Eric Widmaier, Linda Graham, and Peter Stiling. (2014). The Chemical Basis of
Life II: Organic Molecules. Biology 3rd Edition, 42-64.

Hoobler, Cynthia, Karen Duston, Adam Eiler, Jennie Plunkett, Kirsten Raines, and Mary
Wisgirda. Chemical Composition of Cells. (2007). General Biology I and II, 13-21.

– 37 –
 Lab #5 – Microscopy

Lab #5 Microscopy
Introduction

Scientists use microscopes to see objects that are too small to be seen with the unaided eye.
Microscopes fall into two general categories – light microscopes and electron microscopes.

Light microscopes use light and a series of glass lenses to magnify the organisms being viewed.
Magnification generally ranges from 40x to a high of 1000x. At 1000x, however, special oil must
be placed on the slide for adequate resolution. Because of this, we will never use the 1000x
magnification in this lab. Organisms observed may be dead (prepared slide) or living. A wet
mount preparation (to be discussed later) enables observation of living organisms. A subset of
the light microscopes often used in college biology laboratories is the dissection microscope.
As the name implies dissections can be performed on organisms (such as a mosquito) which
would be too small to see otherwise. Details of plants (for example, guard cells in plant leaves)
and insects can readily be seen with a dissecting microscope.

Human tissues viewed under a light microscope

Electron microscopes use a beam of electrons to magnify the organisms being viewed. The two
general categories of electron microscopes are the transmission electron microscope and the
scanning electron microscope. A transmission electron microscope sends a beam of electrons
through a very thin section, or slice, of the specimen. If a scientist wants to see minute details
of cellular organelles, a transmission electron microscope with magnifications approaching
100,000x is ideal. Scanning electron microscopes bounce a beam of electrons off the surface of
the specimen. They are useful to observe cell surfaces and very small organisms. Unlike the
light microscope, however, specimens must be dead before they can be viewed with electron
microscopes.

– 38 –
 Lab #5 – Microscopy

A) picture of scanning electron microscope B) scanning electron microscope image of a flea C) picture of an
electron scanning microscope D) Transmission electron microscope image of human pancreatic cell

Pre-Lab Questions

1. Which microscope uses a series of glass lenses to magnify the organisms being viewed?

2. Which microscope is used to view living organisms?

3. Name the two types of electron microscopes:

4. Which electron microscope is used to view surface features of cells?

5. What is the purpose of today’s lab?

– 39 –
 Lab #5 – Microscopy

6. Are you allowed to have food and drink in the laboratory?

7. How many hands should you use when carrying a microscope around the classroom?
Where should they be located?

8. What two things should you use to clean a microscope?

Materials

Microscope lab handout

Microscope
Plant Root slide
Slides
Coverslips
Protoslo™ or Detain™
Pond water
Lens paper
Lens cleaning solution

Lab Procedures and Data

Rules for Microscope Use

1. When carrying the microscope, one hand should be on the base and the other on
the arm.
2. Use only lens paper and lens cleaning solution when cleaning objectives.
3. Do not allow moisture to get into the inner parts of the microscope.
4. Do not tilt microscope when viewing a wet mount.
5. Scanning power objective should be in the downward position at close of the lab.
6. Turn off the light when you are finished using the microscope.
7. Report any malfunctions to your instructor.

– 40 –
 Lab #5 – Microscopy

Parts of the Microscope

Use the diagram below to label all the parts of the microscope:

– 41 –
 Lab #5 – Microscopy

Give the function of the following microscope parts:

Arm

Body Tube

Nosepiece

Ocular lenes

Objective lenses

Stage

Stage clips

Large stage control knob

Small stage control knob

Coarse adjustment knob

Fine adjustment knob

Filter Slot

Condenser

Diaphragm control

– 42 –
 Lab #5 – Microscopy

How to Use a Microscope

1. Make sure the slide is firmly held in place by the stage clips. The stage clips should touch
the edges of the slide, not be on top. Once it is in place, use the stage control knobs to
move the slide; do NOT move it with your fingers.

2. ALWAYS BEGIN VIEWING ON THE LOWEST POWER OBJECTIVE (4x).

3. Use the stage control knobs to move the slide around until you see something of
interest. Use the COARSE adjustment knob to bring the object into focus.

4. If you want to zoom in on the object (increase magnification), first make sure the object is
at the tip of the pointer on the microscope. If you do not see the pointer, look in the other
ocular or use your other eye. Then increase the magnification by moving to a different lens.
Our microscopes are parfocal, meaning that all the objective lenses focus to the same point.
So, once you have focused on an object it should stay in focus as you change the
magnification. However, at higher magnifications you may still need to make slight
adjustments using the fine adjustment knob.

5. IMPORTANT: Never use the coarse adjustment knob at higher magnifications – you may
damage the microscope.

Magnification
In this lab we are using compound light microscopes, which means that the images we will be
viewing will be magnified twice – once by the objective lens and once by the ocular lens (found
in the eye piece). In these microscopes you can choose to use one of several different
objectives, but you cannot change out the ocular lens. The total magnification is calculated by
multiplying the magnification of the two lenses together. For example, if you are viewing a
specimen with a 5x ocular lens and a 10x objective, the total magnification is 5 x 10 = 50x total
magnification.

Use your microscope to complete the following table.

Objective lens Ocular lens
Objective Total magnification
Magnification Magnification
Scanning
(red line)
Low power
(Yellow Line)
High power
(blue line)
Oil immersion
(white line)

– 43 –
 Lab #5 – Microscopy
*** this objective is NOT used in BIOL I ***
Depth of field
Using any of the slides provided examine the slide at 40X total magnification. Move the stage
so that the tip of the pointer is in the middle of the object, then increase the total
magnification to 100X. Slowly focus up and down using the fine adjustment knob to see how
the image changes at different depths of field. Re-center the object and repeat at 400x total
magnification.

What do you notice as you change the focus?

Making a Wet Mount

A wet mount is a quick and easy way of viewing living specimens on a light microscope. The slide
is made with just enough liquid to allow the specimen to move around on the slide, but not so
much that liquid escapes to contaminate the inner workings of the microscope.

To make a wet mount:

1. Obtain a clean slide and cover slip from your instructor. To avoid contaminating the slide with
your skin oils, handle the slide and coverslip by the edges as much as possible.

2. Place one drop of the liquid or suspension you want to view (e.g. pond water) on the slide.

3. Add one drop of Protoslo™ or Detain™ to the slide. (These are high density chemicals used to
slow down the movement of fast-swimming protists.)

4. Depending on what you are viewing, you may also need to add a drop of stain to make the
different parts of the specimen show up better (follow the instructors directions).

5. Once all the necessary drops are on the slide, slowly lower the coverslip. It is best to start with
the coverslip at an angle so that only one edge is touching the slide, then to slowly lower it the
rest of the way down. Try to have as few air bubbles as possible.

Since living specimens need air to live (even those that swim), many times the best place to find the
specimens is to look around the edges of the coverslip. Remember, always begin your search on the
lowest power objective, then slowly move up to higher magnifications.

– 44 –
 Lab #5 – Microscopy
Observe and attempt to identify the organisms.
Use the space below to draw some of the microorganisms you see in the pond water. Make sure to
record what total magnification you are using to view the specimens (they may look different at
different levels of magnification). Use the posters on the walls of the lab to identify these pond
microorganisms. Sketch, identify, and show to your instructor two different organisms.

Post-Lab Cleanup

Make sure all labeled, prepared slides are returned to the appropriate slide box. Return the
microscope to the CORRECT spot in the cabinet (with the 4x objective in the viewing position). All
slides used in making wet mounts should be put in the buckets at the back of the room, and all used
cover slips should go in the trash. Ethanol the desks and push in the chairs.

Post-Lab Review Questions

1. If the scanning power objective lens is in viewing position, what is the total magnification?
_________
2. What is the function of the diaphragm control? ______________________________________
3. What is the function of the condenser? ____________________________________________
4. Which stage control knob moves the stage from left to right? ___________________________
5. Why do we use DetainTM? _______________________________________________________
6. May you tilt the microscope when viewing a wet mount? ______________________________
7. What is meant by parfocal? ______________________________________________________
8. What objective lens will you never use in this course? _________________________________
9. What are the two types of lenses found in a compound microscope? _____________________

References

Brooker, Robert, Eric Widmaier, Linda Graham, and Peter Stiling. (2014). General Features of
Cells. Biology 3rd Edition, 66-96.

Hoobler, Cynthia, Karen Duston, Adam Eiler, Jennie Plunkett, Kirsten Raines, and Mary
Wisgirda. Microscopy. General Biology I and II, 23-32.

– 45 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Lab #6 Prokaryotic Cells and Eukaryotic Animal Cells

Introduction

Cells can be divided into two major categories – prokaryotic and eukaryotic.

Prokaryotic cells are simple cells that do not have internal membranes. These cells have DNA
within a nucleoid region which directs cellular activity, but it is not membrane bound.
Organelles (small structures within the cell that have specialized function) are lacking in
prokaryotic cells. Bacteria, such as those which cause disease, have prokaryotic cells.

In contrast to prokaryotic cells, eukaryotic cells are quite complex. They have a membrane
bound nucleus that serves as the “brain” of the cell and numerous other membrane-bound
organelles that carry out many functions within the cell. The mushrooms you eat, the trees
that provide you shade, your pet cats and dogs, and you are all composed of eukaryotic cells.

typical prokaryotic cell typical eukaryotic cell

Pre-Lab Questions

Using the information in your textbook, give the function of each of the following organelles
and cellular structures:

Plasma membrane

Nucleus

Nucleoid region

– 46 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Smooth endoplasmic reticulum

Rough endoplasmic reticulum

Ribosomes

Golgi apparatus

Mitochondria

Lysosome

Centriole

Cytosol/cytoplasm

What is the purpose of this lab?

Materials

Models: animal cell, Euglena, Paramecium, Amoeba

Sterile swabs
Glass slides
Coverslips
Methylene blue (stain)
Anabaena slide
Amoeba slide
Paramecium slide
Live cultures of Anabaena, Amoeba, Euglena, and Paramecium (if available).
Protoslo™ or Detain™
Microscope
human epithelial cells slide

– 47 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Lab Procedures and Data

Review the “Making a Wet Mount” section of Lab #5 – Microscopy before beginning. We will
be using the same procedure to create slides of living protists and animal cells in this lab.

Prokaryotic Cells

Anabaena is a genus of bacteria that contains several different

species. All Anabaena are photosynthetic, meaning they can get
energy from sunlight. This means Anabaena are autotrophs –
they can make their own food. Members of this genus are
found all over the world, usually as a type of algae or plankton.

We will be looking at both prepared and stained specimens of

Anabaena and living (wet mount) specimens.

Prepared slide of Anabaena

Anabaena is best viewed at 400x total magnification.
(Remember, always start at with the 4x scanning objective until
you find a specimen on the slide, then zoom in step-by-step!)
Use the space below to sketch a filament (line) of Anabaena Anabaena spherica
cells. Label the prokaryotic cells.

Do these cells have a nucleus?

– 48 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Wet mount of Anabaena (if available)

Sketch a filament of the living Anabaena below. A filament is a chain of cells. They often coil
up, looking like a string of beads.

What differences do you see between the living and prepared Anabaena?

What type of taxonomic classification is the name Anabaena (kingdom, genus, species, etc)?

Are Anabaena prokaryotic or eukaryotic?

To what domain does the organism Anabaena belong?

Eukaryotic Cells

Eukaryotic cells are divided into four different kingdoms: Plant, Animal, Fungi, and Protists.
Today we will examine specimens from the Protist and Animal kingdoms; Lab 7 will examine
specimens from the plant kingdom.

Protists are a loosely defined and incredibly diverse group of microorganisms that live in moist
habitats and are usually single-celled. Some form algae and plankton, others can cause human
diseases. We will examine three representative types of protists today: Paramecium, Euglena,
and Amoeba.

We will also examine human epithelial cheek cells. Epithelial cells are any cells that form
surface tissues – i.e. skin and the lining of any body cavities. We will use cheek cells because
they are easily obtainable without the use of a scalpel.

– 49 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Paramecium
Paramecium is a genus of unicellular protists that are covered in tiny
cilia, which allow them to move through the water. Paramecia are
heterotrophs – they must eat other organisms for energy. They eat
by endocytosis of food particles (including bacteria) through a deep
oral groove. Like most protists, they use contractile vacuoles to get
rid of any excess water they absorb from their environment.
Paramecia are usually between 50 and 350 µm long and are found
mainly in freshwater, though some species have been identified in
saltwater environments.

Model of Paramecium
Use this figure to identify the following structures: macronucleus, Paramecium
micronucleus, oral groove, food vacuole, contractile vacuole, cilia.

– 50 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Prepared Slide of Paramecium

Paramecium is best viewed at 100x or 400x total magnification, though some structures are
easily seen at 40x total magnification (these are large protists). Use the space below to draw a
picture of a Paramecium.

Label the following structures on your sketch: macronucleus, contractile vacuoles, cilia, and
oral groove (NOTE: All of these are structures we could point at and ask you to identify on a lab
practical!)

Wet Mount of Paramecium (if available)

Sketch a living Paramecium below.

What differences do you see between the living and prepared Paramecium?

Are Paramecium eukaryotic or prokaryotic?

Do Paramecium have a nucleus?

– 51 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Euglena
Euglenas are a species of photosynthetic protists found in both salt
and fresh water. Like plant cells, Euglenas have chloroplasts and
are photosynthetic; this makes them autotrophs. They move by
using a flagellum, a long whip-like organelle. Euglenas are often
studied in laboratories as a model organism (an organism that
survives well in the lab and which can be studied to learn about a
variety of other similar organisms).

Model of Euglena
Use this figure to identify the following structures: flagella, eyespot,
Euglena stained to show
contractile vacuole, nucleus, chloroplasts their flagellum

– 52 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Prepared Slide of Euglena

Euglena is best viewed at 400x total magnification if you want to see the chloroplasts. Use the
space below to draw a picture of a Euglena.

Label the following structures on your sketch: nucleus and flagellum.

Wet Mount of Euglena (if available)

Sketch a living Euglena below.

Are Euglenas eukaryotic or prokaryotic?

Are Euglenas protists, animal, or bacteria?

– 53 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Amoeba

Amoebas are a genus of “animal-like” protists –

they hunt and “trap” their food before they eat it.
Amoebas move by using pseudopodia (“pseudo” =
fake and “pod” = feet), long extensions of the
cytoplasm that allow them to creep along a surface in
search of food. Amoebas can even wrap these
pseudopodia around prey (like bacteria or even
paramecia) to bring it inside the cell to eat.

Model of Amoeba
Use this figure to identify the following structures: Hungry Amoeba in search of food
pseudopodium, nucleus, food vacuole, and contractile vacuole.

– 54 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Prepared Slide of Amoeba

Amoeba is best viewed at 100x or 400x total magnification. The pseudopodia are best seen at
100x, but some cellular organelles can only be seen at 400x magnification. Use the space below
to draw a picture of an Amoeba.

Label the following structures on your sketch: pseudopodium, nucleus, food vacuole, and
contractile vacuole.

Wet Mount of Amoeba (if available)

Sketch a living Amoeba below.

Do Amoeba have a nucleus? Are they prokaryotic or eukaryotic?

Are Amoeba protists, animal, or bacteria?

– 55 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Animal cells (human epithelial cells)

All animal cells have different types and functions

of organelles; their specific make-up fits their
function. For example, liver cells have lot of
smooth endoplasmic reticulum because it is used
to break-down toxins, while muscle cells have lots
of mitochondria because they need the energy.
However, the model we will use in this class
shows the most common structures found in
almost all animal cells.

Human epithelial cell photographed

using phase contrast microscopy

How to Make a Wet Mount of Human Epithelial Cells:

1. Use a sterile swab to rub the inside of your cheek. Do NOT stab the inside of your cheek.
We are not looking for blood cells.

2. Rub the swab over the middle section of a clean slide. The more you rub, the better
you will spread out your cells. Blobs are bad.

3. Add one drop of Methylene blue stain to your slide. Warning: Methylene blue stains
skin cells, so it will stain your fingers if you are not careful.

4. Carefully place a coverslip over the stained area. It is best to start with the coverslip at an
angle so that only one edge is touching the slide, then to slowly lower it the rest of the
way down. Try to have as few air bubbles as possible.

5. Note: Sometimes you will see small black specks in a wet mount of cheek epithelial cells.
These are small deposits of tar often found in the cheeks of smokers.

– 56 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Prepared Slide of Human Epithelial Cells

Human epithelial cells are best viewed at 400x total magnification to see the interior structures.
Use the space below to draw a picture of a human epithelial cell.

Label the following structures on your sketch: nucleus, cell membrane, and cytoplasm.

Wet Mount of Human Epithelial Cells

Sketch your wet mount of human epithelial cells below.

Are these cells prokaryotic or eukaryotic?

Why do you not see any cellular structures besides a nucleus in these cells?

– 57 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Model of a Typical Animal Cell

Use this figure to identify the following structures: plasma membrane, lysosome,
mitochondria, ribosomes, rough endoplasmic reticulum, smooth endoplasmic reticulum,
Golgi body, centrioles, secretory vesicle, nuclear membrane, nuclear pore, nucleolus, and
chromatin.

– 58 –
 Lab #6 – Prokaryotic Cells and Eukaryotic Animal Cells

Post-Lab Cleanup

Make sure all prepared, labeled slides are returned to the appropriate slide box. Return the
microscope to the CORRECT spot in the cabinet (with the 4x objective in the viewing position).
All slides used in making wet mounts should be put in the buckets at the back of the room, and
all used cover slips should go in the trash. Ethanol the desks and push in the chairs.

Post-Lab Review Questions

1. Differentiate between prokaryotic and eukaryotic cells. __________________________

_______________________________________________________________________
2. Do the cells in Anabaena have a nucleus? _____________________________________
3. Are Paramecium a protist or a bacteria? Give one piece of evidence for your answer.
_______________________________________________________________________
4. How do smooth and rough endoplasmic reticulum differ? ________________________
5. Which of the cells that we looked at today had a nucleus? ________________________
6. What was unique about the nucleus of the Paramecium? ________________________
7. Which type of cell has a nucleoid region? _____________________________________
8. Which cells that we looked at today were heterotrophs? _________________________
9. Which cells that we looked at today were autotrophs? __________________________

References
Brooker, Robert, Eric Widmaier, Linda Graham, and Peter Stiling. (2014). General Features of
Cells. Biology 3rd Edition, 66-96.

Hoobler, Cynthia, Karen Duston, Adam Eiler, Jennie Plunkett, Kirsten Raines, and Mary
Wisgirda. (2007). Prokaryotic and Animal Cells. General Biology I and II. 33-40.

– 59 –
 Lab #7 – Eukaryotic Plant Cells

Lab #7 Eukaryotic Plant Cells

Introduction

Plants have many of the cellular structures of animal cells, but with some significant
differences. A prominent feature is a large central vacuole which may contain water and other
substances. Plants have cell walls which provide rigidity and protection. Within plant cells are
several unique organelles. These include the chloroplast which contains the pigment
chlorophyll. Chloroplasts allow plant cells to carry out photosynthesis; a series of chemical
reactions whereby the plants extract energy from sunlight. This means the plants are
autotrophs – they can make their own food. The pigment carotene is found in chromoplasts.
Amyloplasts contain starch which appears blue-black when iodine is added to a potato wet
mount.

In this lab, we will be focusing on the structures that are unique to plant cells. You will need to
be able to determine whether a cell is a plant or an animal cell. This lab will give you the tools
to do so.

Pre-Lab Questions

Use your textbook to answer the following questions.

1. Give the function of each of the following cellular structures.

Chloroplast

Central vacuole

Cell wall

Chromoplast

Amyloplast

Plasmodesmata

2. Do plants have centrioles?

3. What is the purpose of this lab?

4. What safety precautions should be taken today?

– 60 –
 Lab #7 – Eukaryotic Plant Cells

Materials

Onion (prepared slide and fresh)

Elodea (prepared slide and fresh)
Carrot
Flower petal
Potato
Glass slides
Coverslips
Microscope
Water
Knife
Cutting board

Lab Procedures and Data

Review the “Making a Wet Mount” section of Lab #5 – Microscopy before beginning. We will
be using the same procedure to create slides of living plant cells in this lab.

Make wet mounts of the following tissues. Use a clean slide, water, and new coverslip for each slide.

Onion Epidermal Cells

Prepared Slide of Onion (genus name Allium)

Slides of onion epidermal cells are best viewed at 100x total magnification. These samples have been
stained to show the cell walls and nuclei.

Use the space below to sketch several onion cells. Label the cell wall, cytoplasm, and nucleus.

Are these cells prokaryotic or eukaryotic?

– 61 –
 Lab #7 – Eukaryotic Plant Cells

Wet Mount of Onion Cells

To make a wet mount of onion cells:

1. Carefully remove one layer of the onion from the bulb.

2. Peel away the thin membrane from the inside curve

of the onion layer (use your fingernails).

3. Place a small piece of this thin layer on the slide

4. Add one drop of iodine to the onion sample.

5. Place a coverslip over the sample.

Sketch several of the living onion cells below. Label the cell wall, cytoplasm, and nucleus.

What differences do you see between the living and prepared onion cells?

What is the genus name for Onions?

To what domain do onions belong?
To what kingdom do onions belong?

– 62 –
 Lab #7 – Eukaryotic Plant Cells

Elodea

Elodea is a type of seaweed usually found in freshwater

environments. It is often used as aquarium decorations and
as a model species. We will use it in many different labs
this semester. Though it is native to North America, it has
invaded several other continents and is considered an
invasive species.

Make a wet mount of an Elodea leaf. The leaves are thin

enough that you can simply place one on the slide, add a
drop of water, and place a cover slip over it. Elodea is best
viewed at both 100x total magnification (to see the cell wall)
and 400x total magnification (to see the chloroplasts). The
chloroplasts of the Elodea leaf are where the chlorophyll
pigment is stored and where photosynthesis takes place (as
mentioned in the introduction to this lab).

Sketch several Elodea cells. Label the cell wall, cytoplasm, and chloroplasts.

What color are the chloroplasts inside the cells?

What chemical is inside the chloroplasts making them that color?

Are the chloroplasts moving inside the cell? What could be causing this?

– 63 –
 Lab #7 – Eukaryotic Plant Cells

Flower Petal Cells

Flower petals are used to attract pollinators

to the flowers of plants so that they will
gather and spread pollen. Several different
pigments can be found inside the central
vacuole of flower petal cells. One of the most
common of pigments is called anthocyanin.
There are over 400 different types of
anthocyanins, which produces red, purple,
and blue flowers. These pigments are most
often stored in central vacuoles of the petal
cells. White flowers do not have any pigments
in their cells.

Tear off a small section of a flower petal and make a wet mount. For best results focus on the
cells near the torn edge. Use 400x total magnification to see the cellular structures.

Sketch several cells. Label the cell wall and central vacuole.

What color are these cells?

What pigment is producing the color in these cells?
Where is anthocyanin stored in the plant cell?
How many central vacuoles are there per plant cell?
Describe what the central vacuole looks like under the microscope. How would you recognize it
if you saw it again?

– 64 –
 Lab #7 – Eukaryotic Plant Cells

Carrot cells

Carrots are known for their orange color (though they

can actually come in a variety of shades). This color is
produced by a pigment called carotene. In plants this
pigment is important for photosynthesis, but in humans
it is broken down to form retinol, a form of Vitamin A.
Vitamin A is very important for proper functioning of
the eye retina.

Cut a very, very thin, transparent slice of carrot (it

should be the thickness of a piece of paper). Make a
wet mount of this thin slice in water. The carrot cells
are best viewed at 100x total magnification at the edge
of your slice.

Use the space below to sketch several carrot cells.

Label the cell wall, cytoplasm, and chromoplast.

What color are these carrot cells? Is the color spread throughout the cells?

What is the name of the pigment stored in these cells?

Where is the pigment stored in the cell?
Roughly how many chromoplasts are there per plant cell?
How is the storage of this pigment different from the anthocyanin storage in the flower petal
cells?

– 65 –
 Lab #7 – Eukaryotic Plant Cells

Potato Cells

Potatoes are a form of long-term energy storage for

plants. Potatoes store starch, which is a
polysaccharide made of long, unbranched chains of
glucose. During the winter, the plant will break down
the starch into individual glucose monomers that can
be used to fuel cellular respiration and provide energy
to the plant. Alternatively, the potatoes can be eaten
by humans who can also break down the starch and use the glucose for energy. Potatoes are
the world’s fourth-largest food crop (behind rice, wheat, and corn).

Cut a very, very thin, transparent slice of potato (it should be the thickness of a piece of paper).
Make a wet mount of this thin slice in water. The potato cells are best viewed at either 100x or
400x total magnification at the edge of the slice.

Use the space below to sketch a few potato cells. Label the cell wall, cytoplasm, and
amyloplast.

Carefully remove the coverslip from your potato wet mount slide and add a drop of iodine to
the slice of potato. Use the space below to sketch the appearance of the potato cells after
expose to iodine.

How did the appearance of the cells change after you added iodine?

What chemical is stored in these potato cells?

Explain why the color change you observed took place (Hint: Refer to Lab#4).

Which cellular organelle stores the chemical found in these cells?

– 66 –
 Lab #7 – Eukaryotic Plant Cells

Model of a Typical Plant Cell

Use this figure to identify the following structures: cell wall, plasmodesmata, plasma
membrane, mitochondria, ribosomes, rough endoplasmic reticulum, smooth endoplasmic
reticulum, Golgi body, central vacuole, chloroplasts, nuclear membrane, nuclear pore,
nucleolus, and chromatin.

– 67 –
 Lab #7 – Eukaryotic Plant Cells

Post-Lab Cleanup
Make sure all prepared and labeled slides are returned to the appropriate slide box. Return the
microscope to the CORRECT spot in the cabinet (with the 4x objective in the viewing position).
All slides used in making wet mounts should be put in the buckets at the back of the room, and
all used cover slips should go in the trash. Large pieces of unused plant materials should be left
at on the lab bench; small pieces should go in the garbage. Ethanol the desks and push in the
chairs.

Post-Lab Review Questions

1. Name the pigment inside the chromoplast. ____________________________________________

2. Name the pigment inside the chloroplast. _____________________________________________
3. In the flower petal, the pigment anthocyanin is contained in what structure? _________________
4. Name the substance inside the amyloplasts. ___________________________________________
5. What color are amyloplasts after the addition of iodine? Why? ____________________________
________________________________________________________________________________
6. Name three organelles found in plant cells but not in animal cells. __________________________
________________________________________________________________________________
7. For each of the following, state whether this is a characteristic of a plant cell, animal cell, or
protist and why.
a. Rectangular shape _____________________________________________________________
b. Cilia_________________________________________________________________________
c. Pseudopodia_________________________________________________________________
d. Green structures inside it _______________________________________________________
e. Orange structures inside it ______________________________________________________
f. Oral groove __________________________________________________________________
g. Food vacuole _________________________________________________________________
8. Compare how the colorful pigments are stored in flower petal cells vs. carrot cells. ____________
________________________________________________________________________________

References
Brooker, Robert, Eric Widmaier, Linda Graham, and Peter Stiling. (2014). General Features of
Cells. Biology 3rd Edition, 66-96.

Hoobler, Cynthia, Karen Duston, Adam Eiler, Jennie Plunkett, Kirsten Raines, and Mary
Wisgirda. (2007). Plant Cell Structure. General Biology I and II. Boston: McGraw-Hill, 41-44.

– 68 –
 Review for Lab Practical #1

Review for Lab Practical #1

How a lab practical works
Here are some things you need to know in order to prepare for a lab practical in General
Biology I or II. 30 stations will be set up, so each student has his/her own spot to start at.
There will be an index card with 2 questions on it -- so there is a total of 60 questions on the
exam. Students stand, not sit, to take the exam. That is why the chairs have been pushed out
of the way.

Lab practicals are not multiple choice. Answer sheets will be provided. Answers are usually a
single word or short phrase (food vacuole, nucleus, fine focus, etc.).

Examples of stations:
 A microscope with a slide, label covered, focused, pointer pointing at a structure on
the slide.
 A model or specimen with a piece of tape or a numbered pin attached to a structure.
 A test tube rack with test tubes containing solutions that represent the outcome of
the experiment.
 A piece of equipment that was used in an experiment.

The material for exam questions (including Critical Thinking questions) is derived from the
lab manual. Students are responsible for knowing everything covered by the lab manual.

In Bio I, many questions are about the materials and equipment used and the outcome of
the experiment, asking the student to interpret the results. Students will not be asked to
perform experiments or find specimens on microscope slides. For example, questions about
the lab on Chemical Composition of Cells might include:
 Name the reagent used to test for the presence of proteins.
 Of the two test tubes here, which one shows a positive test for proteins?
 Of the two test tubes here, which one is the
control? Questions about the Paramecium might include:
 Name this organism.
 Name the structure at the tip of the pointer (macronucleus, contractile vacuole, etc.).

The lab practical is timed with a stopwatch. When the instructor says “Move”, all students
move to the next station at the same time. There will be 10 minutes at the end of the exam to
go back to stations to recheck questions. Only one student is allowed per station at a time.

– 69 –
 Review for Lab Practical #1

To prepare for Lab Practical I: use lab manual and this review! Hint: Pre-lab questions
make excellent lab practical questions!

Metric measure

Know how to do metric conversions. Be able to convert between cm, mm, µm, and nm.

Practice Conversion Problems (Answers at the end of the review.)

1 cm = mm 1 mm = µm 1 µm = nm

1mm = cm 1 µm = mm 1 nm = µm

23 cm = mm 3 nm = µm 368 mm = µm

185 mm = cm 6 µm = nm 0.7 mm = µm

Length
Using a metric ruler, be able to measure the length of a line.
Using a meter stick and wooden paddles, be able to measure the length of a bone.
Be able to give your answers in either centimeters or millimeters.
On the practical, be sure to write the correct unit of measure beside your answer.

Temperature
Know how to read a thermometer and properly record the results.
Do Biol I thermometers read in Celsius or Fahrenheit?
Which is the metric unit for temperature?
Memorize: freezing point of water in both oC and oF
boiling point of water in both oC and oF

Weight
Be able to read the weight of an object on the triple beam balance.
Be able to convert g to mg and mg to g. Example: 32 g = mg or 32 mg = g

Volume
Be able to read the volume of water in a graduated cylinder (units: ml)
Term to know: meniscus
Which part of the meniscus is used to read the volume of water in a graduated cylinder?

pH and Buffers

Be able to determine the pH of a substance using pH paper and the colored scale.
Know if the pH number indicates presence of an acid or a base.
pH greater than 7 is a pH less than 7 is a

– 70 –
 Review for Lab Practical #1
Is HCl an acid or a base? What gives you the clue?
Is NaOH an acid or a base? What gives you the clue?
Be able to identify: a common substance we tested that is an acid.
a common substance we tested that is a base.

Buffers
Know the pH of distilled water.
Know what happens to the pH of distilled water when acid is added.
Know what happens to the pH of a buffered solution (pH 7) when acid is added.

Presented with two tubes: one tube with a buffered solution + acid and one tube with water +
acid, how will you know which tube has the buffer and which tube does not have the buffer?

Presented with a graph similar to the one you drew in lab, be able to interpret which buffered
solution was the best at maintaining the pH of a solution

Effectiveness of Antacids
Name the pH indicator used in this experiment
What color is the antacid solution when the pH indicator is added?
What color is the antacid solution/pH indicator when the antacid is no longer effective?
Presented with two test tubes, one has a yellow substance, and one has a purple
substance,
What is the color of the tube with the lower pH number?
What is the color of the tube with the higher pH
number?

Be able to predict which tube indicates the more effective antacid.

Name an antacid we tested in lab that was very effective.

Molecular Models

Know the atom each colored ball represents: red yellow black blue
Know what type of bond the sticks and springs represent. Be specific!
Given a molecular model, be able to determine whether the molecule is organic or inorganic.
Given a molecular model, be able to write the empirical formula of the molecule.

– 71 –
 Review for Lab Practical #1

Be able to identify the following molecular models and functional groups:

Molecules Functional groups
water amino acid: glycine amino group
ethyl alcohol amino acid: alanine carboxyl or acid group
glucose dipeptide hydroxyl group
ribose “R” group
deoxyribose
fructose
disaccharide

How many monosaccharides make up a disaccharide? What molecule is removed to make it?
How many amino acids make up a dipeptide? What molecule is removed to make it?

Chemical Composition of Cells

Testing for protein:

What reagent is used to test for protein?
Presented with test tubes of substances with biuret added to each tube:
What color indicates the presence of protein?
Is this color a positive or negative test result for protein?
What color indicates the absence of protein?
Is this color a positive or negative test result for protein?
In this experiment, what substance was used as a control?

Testing for starch:

What reagent is used to test for starch?
Presented with test tubes of substances with iodine added to each tube:
What color indicates the presence of starch?
Is this color a positive or negative test result for starch?
What color indicates the absence of starch?
Is this color a positive or negative test result for starch?
In this experiment, what substance was used as a control?

Testing for monosaccharides (simple sugars):

What reagent is used to test for monosaccharides? Be specific!!
Presented with test tubes of substances with Benedict’s solution added and each tube boiled:
What color(s) indicates the presence of monosaccharides?
Is this color a positive or negative test result for monosaccharides?
What color indicates the absence of monosaccharides?
Is this color a positive or negative test result for monosaccharides?
In this experiment, what substance was used as a control?

Testing for lipids:

– 72 –
 Review for Lab Practical #1
How did we test for presence of lipids?
Describe the appearance of the paper towel when a lipid is present.
Is this a positive or negative result?
Describe the appearance of the paper towel when a lipid is not present.
Is this a positive or negative result?
In this experiment, what substance was used as a control?

Microscopy

Know the parts of the compound light microscope we use in lab.

Know the function of each part.

Know how to determine the total magnification when using different objective lenses.

Why is the word compound used to describe our microscope?

Be able to name the two types of lenses in our compound scope.
What is meant when it is said that our microscopes are parfocal?

Prokaryotic Cells

Be able to recognize Anabaena when looking through the microscope.

Recognize Anabaena on the prepared slide and on a fresh mount of Anabaena.
Know that Anabaena is a type of bacteria.
Do the cells in Anabaena have a nucleus? Are Anabaena cells prokaryotic or eukaryotic?

Eukaryotic Cells

Protists
Know that protists are eukaryotic.
Be able to recognize the following organisms and know these organisms are protists.
Amoeba prepared slide and model
Parts to identify on both the model and slide:
cytoplasm, nucleus, pseudopodium, contractile vacuole, food vacuole

Paramecium prepared slide and model

Parts to identify on model: cilia, oral groove, macronucleus,
micronucleus, food vacuole, contractile vacuole
Parts to identify on slide: cilia, oral groove, contractile vacuole,
macronucleus.

Euglena prepared slide and model

Parts to identify on model: flagellum, eyespot, contractile vacuole,

– 73 –
 Review for Lab Practical #1
nucleus, chloroplast
Parts to identify on slide: nucleus

Animal Cells
Know that animal cells are eukaryotic.
Be able to recognize squamous epithelial cells taken from the inside of the cheek.
Parts to identify: cell membrane, nucleus, cytoplasm

Animal Cell Model

Be able to identify the following organelles.
plasma membrane ribosomes
nuclear membrane mitochondria
nuclear pore lysosomes
nucleolus Golgi body
chromatin centrioles
rough endoplasmic reticulum smooth endoplasmic
reticulum secretory vesicle

Plant Cells

Know that plant cells are eukaryotic.

Be able to recognize the following as plant cells.
Onion epidermal cells (fresh preparation and prepared
slide) Parts to identify: cell wall, cytoplasm, nucleus

Elodea cells (fresh preparation)

Parts to identify: cell wall, cytoplasm, chloroplasts
Know the name for the green pigment inside the chloroplasts.

Carrot cells (fresh preparation)

Parts to identify: cell wall, cytoplasm, chromoplasts.
Know the name of the orange pigment inside the chromoplasts.

Potato cells (fresh preparations - one unstained and one stained with iodine)
Parts to identify: cell wall, cytoplasm, amyloplasts
Know the substance inside the amyloplasts.

Flower petal cells (fresh preparation)

Parts to identify: cell wall, central vacuole
Know the name of the red pigment inside the vacuole.

Plant cell model Be able to identify the following organelles.

cell wall ribosomes
plasma membrane mitochondria

– 74 –
 Review for Lab Practical #1
nuclear membrane Golgi body
nuclear pore chloroplast
nucleolus plasmodesmata
chromatin central vacuole
rough endoplasmic reticulum smooth endoplasmic reticulum

Answers to the practice conversion problems:

1 cm = 10 mm 1 mm = 1000 um 1 um = 1000 nm

1mm = 0.1 cm 1 um = 0.001 mm 1 nm = 0.001 um

23 cm = 230 mm 3 nm = 0.003 um 368 mm = 368,000 um

185 mm = 18.5 cm 6 um = 6000 nm 0.7 mm = 700 um

– 75 –