Name __________________________________________________ Date______________________

Instructor’s signature ______________________

Laboratory Activity 4 – THE PROKARYOTIC CELL

INTRODUCTION

Microorganisms come in considerable variety of shapes, sizes and colonial arrangements. Among all
microorganisms, bacteria are the most studied because they are widely distributed in our environment.
The size of bacteria is expressed in micrometer (µm) and its shape is constant because of its rigid cell wall.
Thus, the basic classification is on the basis of their morphology (form). Bacteria have three basic shapes:
spherical (cocci/ coccus); rod (bacilli/ bacillus); and spiral (spirilli/spirillum).

OBJECTIVES
1. To list the structures present in a prokaryotic cell,
2. To distinguish a prokaryote from other cell types,
3. To observe live specimens and see them in their natural behavior.

MATERIALS
 Compound microscope
 Prepared slides of bacteria (Staphylococcus, Bacillus)
 Concave slide
 Yogurt
 Charts or models of prokaryotic cells

CONTENT

A prokaryotic cell is a type of cell that has no membrane-bound nucleus (pro = before; karyo = nucleus).

Prokaryotes have the following characteristics:

1) Simplest of the organisms
2) Cells are small; cytoplasm is surrounded by a plasma membrane that is enclosed in a cell wall
3) Cells have no distinct interior compartments; no membrane-bound organelles
4) Nucleoid (DNA) is seen as a dense central region apart from the cytoplasm with no nuclear
membrane
5) Contain complex organelles; lack elaborate cytoskeleton but have actin-like proteins

In a prokaryote, the proteins, enzymes, DNA and other molecules are not bounded by a membrane,
therefore, they can be in any part of the cell when needed for chemical reactions. The prokaryote
functions as a single unit. Some prokaryotes are photosynthetic; others are chemosynthetic. They are
important ecologically, have various industrial uses and can also cause diseases.
The simplest method in examining microorganisms is through the “hanging drop” method. This method
allows examination of the characteristics of live cells in relation to its motility, shape and arrangement.
Reproductive activity like binary fission can be also observed in hanging drop method. This technique uses
a concave slide (a slide that has a well in the center) and a cover slip that holds a drop of suspended fluid
with microorganism.

PROCEDURE

A. Hanging Drop Method

1. Get a cover slip and clean it, making it free from grease. In cleaning the cover slip, you may dip it
in alcohol or wash with soap and water then polish it with dry tissue.
2. Get a concave slide by holding it along its edges to prevent contamination with dirt from the finger
tips. Heat the slide to remove the grease and kill some microorganism that may be present. After
heating, place the slide on the table, its concave part is facing up on the table.
3. Place a thin film of petroleum jelly around the ring side of the concavity on the slide.

4. Sterilize the wire loop by holding at 45 degrees angle to the flame. Wait until the wire loop turns
red hot. Then heat up to ¾ of the handle by passing in the flame at the same angle.
5. Take a loopful of culture and place it at the center of the cover slip.
If you’re using a solid culture media, place a drop of normal saline
solution (NSS) on the cover slip before placing the loopful of
bacteria. If you’re using a liquid culture media, no need to place
the NSS, just place the loopful of bacteria directly on the cover slip.
6. Sterilize the wire loop before placing it in the container.
7. Carefully invert the cover slip with bacterial drop and place it over the concavity of the concave
slide. Press it gently so that the petroleum jelly adheres to the cover clip. Make sure that the
bacterial drop is at the center and hanging in the cover slip without touching the concavity of the
slide.

or

8. Place a drop of oil over the cover slip of the prepared specimen and observe the specimen under
oil immersion objective.
Cover slip Petroleum jelly

Microorganism

9. Observe the activities performed by the bacteria such as motility and binary fission.
10. Record your observation in the report sheet.
11. After you observed the slides, discard the slide by placing it a container with disinfectant.
 B. Prepared Slide Viewing
1. Study the typical parts of a prokaryote. Research on the function(s) of each cellular
component and fill in the table below.
2. Examine prepared slides of bacteria at 10x objective; then at 40x, and lastly in 100x.
3. Identify the visible parts. Draw and label accordingly.
4. Compare the structures present in each specimen.

RESULTS AND DISCUSSION:

Hanging Drop Slide Hanging Drop Slide

Total Magnification (TM): __________________ Total Magnification (TM): __________________

Observation: Observation:

Staphylococcus sp. Bacillus sp.

Total Magnification (TM): __________________ Total Magnification (TM): __________________

Observation: Observation:
Provide the function of the typical parts of a prokaryotic cell.

Cellular
Structure Function(s)
component
Double-layer of phospho-lipids
Cell membrane
With proteins & other molecules.

Cytoplasm Gel-like fluid inside the plasma membrane

Ribosomes Granules of nucleic acid and proteins

Genetic material (DNA),

Nucleoid
not surrounded by nuclear membrane

Plasmids Non-essential piece of DNA

Combination of proteins, carbohydrates,

Cell wall
lipids

Present in some bacteria outside the cell

Glycocalyx
wall, slime layers and capsules

Cell extensions Flagella, fimbriae, pili

Figure 1. Typical electron micrograph of a prokaryote

CONCLUSION

QUESTIONS:

1. List five major characteristics unique to prokaryotic cells

a. ________________________________________________________________________
b. ________________________________________________________________________
c. ________________________________________________________________________
d. ________________________________________________________________________
e. ________________________________________________________________________

2. In hanging method, you were able to see microorganisms moving. Differentiate the Brownian
movement and true motility exhibited by bacterial cells in the wet mount slide.

3. Complete the Venn diagram showcasing the differences and similarities between prokaryotic and
eukaryotic cell.

Prokaryotic cell Eukaryotic cell