Name: ________________________________________________ Date:___________

Year and Section: ______________________________________Score:___________

Examination of Living Microorganism (Wet Method)

Direct Wet Mount

Background Information:
In previous activity, the students have learned how to properly use a microscope
and care for a microscope. The microscope is the microbiologist’s basic tool. The
students were able to familiarize and understand each part of the compound microscope.
Students are able to calculate magnification in order to understand to what power they are
observing the specimen.

In order to accurately look at specimens under a microscope, students first need to

learn how to properly prepare a wet mount. A wet mount is made by placing a fluid
solution on a slide, suspending a specimen in a solution, and then covering the specimen
and the solution with a cover slide.

Why would use a wet mount?

• To increase the specimen translucency and to make it easier to stain.
• Using a wet mount slide has the tendency to flatten the specimen making
it easier to view.

Materials:
• dry, clean slide
• cover slips
• canal/dirty water
• pencil
• dropper
• microscope

How to make a direct wet mount:

1. Using a dropper, place a small drop of the canal water at the center of the slide.
2. Carefully place a clean cover glass over the drop.
3. Place your wet mount preparation on your microscope stage, and adjust the
amount of light.
4. Use the low-power objective to locate the microbe.
5. Switch to the high-power objective, focus with the fine adjustment knob, and
adjust the light.
Now using the skill of constructing a wet mount, follow the steps above to help you.

1.) Set up your microscope at your workstation.

2.) Prepare your first wet mount. When it is complete, look at it under the
microscope. Draw what you see.

LPO HPO

3.) What is the benefit of preparing a good wet mount in observing live
microorganisms?

4.) What will happen if you prepare a wet mount with not enough water? Will it
affect your sample? What if there’s too much water?
Hanging Drop Method

Background Information:
Looking at living microorganisms are not as easy as one would think. First of all,
living microorganisms have no color, and they are small: therefore, they are really
difficult to see, even with the oil immersion lens. Second, all microorganisms have some
vibrational movement, even nonmotile ones.
This Brownian movement is caused by water molecules bouncing around in the
solution, knocking up against each other and the microorganisms. Kinetic energy inherent
to all molecules causes this kind of movement. On the other hand, those microorganisms
with flagella will be very apparently moving about the field of vision, although perhaps
not all of the bacteria will be moving. Some cells will "run" straight across the field,
others will "tumble" across the field in a slower motion.

Materials:
• dry, clean concave slide
• cover slips
• canal/dirty water
• petroleum jelly
• dropper
• microscope

How to make a hanging drop wet mount:

1. Place a drop of the canal water in the middle of a cover slip.
2. Place a thin line of petroleum jelly around the edge of the cover slip, or at the four
corners of cover slip.
3. Turn the depression slide upside-down (depressed area facing down) and gently
touch the cover slide. The jelly holds the cover slip to the slide and also keeps the
suspension from drying out.
4. Now flip the entire microscope slide/cover slip combination over. It should look
like the diagram below.
Answer the following:

1.) What is the use of the petroleum jelly in the preparation of the hanging drop
method?

2.) What is the difference between true motility and Brownian movement?

3.) What is the cause of Brownian movement?

CONCLUSION: