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Lab 4: Cell Structure of Prokaryotes, Protists,

Plants and Animals

Name: ________________________________________

Prelab Exercises
Use your textbook or other reliable sources of information to help you answer the
following questions. You need to turn in this prelab exercise immediately upon entering
the lab session.

1. Bring your Biology text or ebook (laptop or tablet—not phone) to lab this week.
Chapters 6, 27 and 28.

2. You are given 4 beakers containing solutions of sucrose sugar in water. Solution A
is 25% sucrose, solution B is 10% sucrose, solution C is also 10% sucrose, and
solution D is 5% sucrose. Compare each solution to the other three solutions using
the terms isotonic, hypertonic, and hypotonic. Please write your answer in complete
sentences. (1pt)

3. Describe how prokaryotic cells differ from eukaryotic cells. (0.2pt)

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4. Describe at least 2 differences between animal cells and plant cells. (0.2pt)

5. List at least 5 membrane-bound organelles you might expect to see in eukaryotic


cells. You should become familiar with these prior to lab by reading about them in
chapter 6. (0.2pt)

6. Protists are eukaryotic organisms. They are often discussed in terms of their
methods and structures involved in locomotion. Fill in the chart with the names of the
organelle(s) that the example organisms use for locomotion. (0.3pt)
Example organisms Name of organelle(s) used for
locomotion

Excavates Euglena and


Euglenozoans Trypanosoma

SAR
Alveolates
ciliates Paramecium

Unikonts Entamoebas (Amoeba)


Amoebozoans and Slime molds

7. Using the information on pages 606-607 explain why systematists favor grouping
protistan red and green algae with land plants instead of with protists. (0.1pt)

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Lab 4: Cell Structure of Prokaryotes, Protists,
Plants and Animals

This week you will be combining many concepts you have learned and skills you
have acquired to make observations of cells. You will be practicing techniques in
microscopy; identifying cell structures you’ve learned about through your textbook and
in lecture, and becoming familiar with the types of cells found in the organisms we’ll
study as we move through the course.

Objectives:
1. Know the difference between prokaryotic and eukaryotic cells.
2. Be able to identify bacilli, spirilli and cocci bacterial shapes.
3. Be able to identify vegetative cells and heterocysts in Anabaena.
4. Be able to identify the representative protist organisms observed in lab.
5. Name the characteristic structures and their function of a plant cell.
6. Name the characteristic structures and their function of an animal cell.
7. Know the difference between the red blood cells of the frog and human.

PROKARYOTIC CELLS: Bacteria and Cyanobacteria

Exercise 1: Bacterial Cell Types


Bacteria are prokaryotic organisms. That is, they lack membrane-bound organelles (like
mitochondria, chloroplasts, Golgi complex, endoplasmic reticulum, nuclei, lysosomes,
etc.). Also, they are much smaller than most eukaryotic cells, so we won’t be able to
see much, if any, of their internal structure.

However, bacteria come in a variety of shapes, which we can see at 400x. Three
especially common shapes are rod, spiral, and spherical. Rod-shaped bacteria are
known as bacilli (singular, bacillus); spiral forms are known as spirilla (singular,
spirillum); and spherical bacteria are called cocci (singular, coccus). Examples of each
of these three common types of bacteria can be found in different regions of the same
microscope slide.

Materials:
Prepared slide “Bacteria 3 types separate smears” (Look at the slide before putting it on
the microscope. NOTE the 3 separately stained areas of the slide)

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Procedure:
1. Examine the slide of bacterial shapes under high power (focus with the 4X, then
10X objective and then switch to 40x objective). Move the slide from side to side
in order to see all 3 types.
2. In the spaces provided, draw each type of bacteria.
3. Record whether these different types of bacteria occur in pairs, chains, clusters,
or as individual, unattached cells.

Bacilli (400X) Cocci (400X) Spirilla (400X)

Exercise 2: Observation of Anabaena, a filamentous cyanobacterium


Anabaena is a prokaryotic organism like bacteria. Anabaena is a filamentous,
photosynthetic, cyanobacterium comprised mostly of photosynthetic cells. Sporadically
interspersed along the chain of photosynthetic cells are cells known as heterocysts.
Heterocysts have the unique ability to convert nitrogen (N2) into ammonia (NH3), a
process known as nitrogen fixation. This conversion is important because most plants
depend on nitrogen for growth, but are unable to use it unless it is converted to NH3.
Soil bacteria are another type of prokaryote that can fix nitrogen. It is not surprising that
soil bacteria have mutualistic relationships with plants.

Materials:
Clean microscope slide and coverslip
Living culture of Anabaena

Procedure:
1. Make a wet mount of Anabaena from the living culture on the side bench.
2. Observe the organism under 100x (using the 10x objective) and 400x and record
your observations. Be sure to label vegetative cells and heterocysts. Figure 27.14
of

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your text.

Label:
Photosynthetic cells
Heterocysts

Anabaena (100 X) Anabaena (400 X)


EUKARYOTIC CELLS: Protists, Animals and Plants

Exercise 3: Observation of Protists


During this lab session, you will begin to appreciate the enormous diversity of eukaryotic
organisms. We’ll begin with single celled eukaryotes called protists. You’ll also look at
some colonial protists and multicellular algae exhibiting greater and greater
interdependence among individual cells.

For each taxonomic group and representative organism, consider its relationship to
other taxa (see figure 28.2 on page 594), how they live (Where do they live? How do
they acquire food?) and consider their structure (What do their cells look like? Are they
unicellular, multicellular, colonial?)

Located on the side bench, you will find several cultures or specimens of representative
protists. They are:

Euglena, a flagellate (Excavata) (page 598, figure 28.8)


Laminaria, a brown alga (SAR, Stramenopile) (pages 600-601, figure 28.13)
Diatoms (SAR, Stramenopile) (page 595-figure 28.2, and page 599)
Paramecium, a ciliate (SAR, Alveolate) (page 604, figure 28.17)
Volvox, a colonial green alga, (Archaeplastida) (page 595-figure 28.2, and pages 602-
604)
Amoeba (Unikonta, Amoebozoan) (page 595-figure 28.2 and pages 609-611)

Materials:
Microscope slides and coverslips
Protoslo or Detain
Petroleum jelly
Living cultures of Paramecium, Euglena, Amoeba, Volvox, and Diatoms
Prepared slide of Laminaria blade with sporangia
Preserved specimen of Laminaria

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Procedures:
For Paramecium and Euglena:
1. Prepare wet mount slides of the living cultures using a small amount of Detain or
Protoslo to slow the organisms. Place a drop of cells and a drop of slowing agent
on the slide. Mix them together with a toothpick (your TA will demonstrate this
procedure).

2. Using your microscope focus with the 4X objective first, then 10X objective and
then switch to 40X objective. Observe the organism under high power. Use the iris
diaphragm to adjust the light.

For Chaos (the Amoebazoan) and the Diatoms:


1. The organisms will be on the bottom of the specimen jar. Using a pipet carefully
suck up some material from the bottom of the specimen jar. Make a wet mount. (Do
not use protoslo or detain)

2. Using your microscope, focus with the 4X first, then the 10X objective. Examine
each organism under medium power. Use the iris diaphragm to adjust the light.

For Laminaria:
1. Obtain a prepared slide of Laminaria blade (leaf like structure of brown algae). The
cells on the surface of the blade develop into sporangia (stained red). The
sporangia develop into zoospores that develop into male and female gametophytes
and then eventually a full grown brown alga.

2. Examine the blade with your microscope. Focus with the 4X objective first, then
10X objective. Use the iris diaphragm to adjust the light.

For Volvox:
1. You will be making a wet mount using petroleum jelly to slightly elevate the
coverslip so that it doesn’t flatten the organism. Your instructor will demonstrate
this technique for you.

2. The Volvox will be visibly suspended in the test tube. Hold the test tube up to the
light to see the green spheres in the tube. Using a pipet carefully suck up a couple
of the spheres. Make a wet mount as shown by your instructor.

3. Using your microscope, find a green dot with the 4x objective first and then move
the dot to the middle of the field and then focus with the 10x objective. Observe the
organism under medium power. Close the iris diaphragm to decrease the light.

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Euglena Laminaria Diatoms
400X 100X 100X
Key Characteristics: Key Characteristics: Key Characteristics:

Paramecium Volvox Chaos (an Amoeba)


400X 40X or 100X 100X or 400X
Key Characteristics: Key Characteristics: Key Characteristics:

Exercise 4: Observation of Plant Cells: Elodea


Two weeks ago, we observed plasmolysis in Elodea plant cells. This week we will look
at Elodea again to observe the structure of a typical plant cell. Elodea is a freshwater
plant commonly found in ponds and freshwater aquaria.

Materials:
Elodea
Microscope slide and coverslip
Water

Procedure:
1. Make a wet mount of an Elodea leaf.
a. Place two drops of water on a clean microscope slide.
b. Gently pluck a leaf from near the tip of a branch of an Elodea plant.
c. Place the leaf bottom-side-up in the water on your microscope slide.
d. Carefully cover the leaf with a cover slip making sure to avoid air bubbles.

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2. Examine the leaf under medium power (10x objective), and then increase the
magnification to high power (40x objective).

3. Study a single cell under high power. Use the fine adjustment and continually
focus up and down to perceive depth. Observe the cell wall completely enclosing
each cell.

The cell membrane adheres to the inside of the cell wall and is probably not visible.
Much of the cell consists of a large, central vacuole containing cell sap (mostly water).
The cytoplasm is present as a thin, nearly transparent fluid layer restricted to the
edges of the cell because of the space filled by the central vacuole. The chloroplasts
are seen as green, oval bodies in the cytoplasm. You may see them passively moving
around the cell as the cytoplasm flows about in what is known as cytoplasmic
streaming or cyclosis. Draw and label the Elodea leaf in the space provided.

The nucleus of the cell may be obscured by the chloroplasts in the cell. If you focus up
and down with the fine adjustment and scan the cytoplasm bordering the cell wall, you
may detect the nucleus as an enlargement in the cytoplasm. You may have to adjust
the amount of light using the iris diaphragm, to see the nucleus or other cellular
components.

Label:
Cell wall
Cell membrane
Chloroplasts
Central Vacuole
Nucleus?

Elodea 400X

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Exercise 5: Observations of Animal Cells – Epithelial cells of the
human mouth
Your mouth is lined with cells known as squamous epithelial cells. Being subject to the
wear and tear of everyday activity in the mouth, these cells are continually sloughed off
and replaced. You can spare a few for the purpose of education.

Materials:
Microscope slide and coverslip
Toothpick
Methylene Blue

Procedure:
1. Add 1 drop of methylene blue to the center of a clean slide.

2. Gently scrape the inside of your cheek with the tip of a toothpick.

3. Roll the tip of the toothpick with the scraping in the drop of methylene blue on the
slide.

4. Cover the drop with a coverslip.

5. Examine the cells by first, finding darker blue dots with the 4x objective. Then move
the dots to the middle of the field and the focus with the 10x objective and then
increase the magnification to high power (40x objective). You may observe some
prokaryotic bacterial cells attached to your eukaryotic animal cell. (Don’t panic, this
is normal.)

6. Draw a cheek epithelial cell as observed under high power, in the space provided.
Label the structures that you can see.

7. Using the field diameter of your 400x total magnification (0.5 mm -- Lab 2), estimate
the size of one of your cheek cells in micrometers.

8. Dispose of toothpick in the regular trash. Dispose of slide in glass disposal.

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Label:
Cell Membrane Nucleus
Nucleolus? Bacteria?
Cheek cell (400 X)
Estimated size _________µm

Exercise 6:
Observations of
Animal Cells –

Frog Blood and Human Blood


Blood is a tissue comprising many types of cells and performing several different
important functions in multicellular animals. The three most basic types of blood cells
are red blood cells, white blood cells and platelets; all of which are suspended in a fluid
called plasma, in which numerous types of molecules are dissolved or suspended.
We’ll be comparing blood from two different animals: frog and human.

Materials:
Prepared slide of frog blood
Prepared slide of human blood

Procedure:
1. Look at the prepared slides of the frog and human blood under high power (first
focus with the 4X, then 10X objective and then switch to 40x objective).
2. Draw and label the blood cells as observed under high power in the spaces
provided. Label the red blood cells and the white blood cells. For both organisms,
the red blood cells will be the most numerous cells in your field of view.

Label:
Red blood cells
White blood cells

Frog Blood (400X) Human Blood (400x)

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How do human red blood cells differ from those of the frog?

Do human red blood cells have a nucleus?

Do all white blood cells look the same?

Clean Up:
1. Clean your microscope eyepieces and lenses with lens cleaner and lens paper. Put
the low power objective down, replace cover, unplug from outlet, and leave on table. 2.
Return the Bacteria, Laminaria, human blood and frog blood slides to the proper slide
trays.
3. Volvox slides made with Vaseline, put the whole slide in the glass disposal.
4. Cheek cell slides put the whole slide in the glass disposal.
5. Coverslips  regular trash or glass disposal – do not put in sink.
6. Microscope slides – wash with warm soapy water, dry and return to the boxes at
your table.
7. Toothpicks, lens paper, paper towels  regular trash.
8. Wipe your table clean with wet paper towels.

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Discussion Questions:
1. What are some benefits that eukaryotes gain from having membrane-bound
compartments within their cells?

2. The endosymbiont or endosymbiotic theory of the history of mitochondria and


chloroplasts (plastids) (pages 532-533 and 593-597 of your text) suggests that these
organelles are descendants of ancestors that were free-living, single-celled
prokaryotic organisms that began living inside larger cells. Over many generations,
they evolved into key organelles in eukaryotic organisms. What evidence supports
this idea? What are their roles in cells?

3. Why is the relationship between surface area and volume of a cell important in
determining cell-size limits?

4. All multicellular eukaryotes (fungi, plants, animals) are descendents of unicellular


protistan ancestors. How do you think multicellular organisms could have arisen
from single celled ancestors? (consider the information from chapter 28 on colonial
and multicellular algae, cellular slime molds, and choanoflagellates)

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5. Why has it been difficult to classify protists?

6. What does it mean to be biologically “successful”? Which groups of organisms have


been the most successful over the history of life on Earth?

As a group, and with each group member contributing, make a diagram (drawing) of a
hypothetical cell that possess all of the eukaryotic cell structures listed in the chart on
pages 100 and 101 in your textbook (although all cells might not actually have all of
these structures. This is a hypothetical cell, not a real one.). As you add each structure
to the cell, make sure each person in the group understands the function of the
structure you are drawing.

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