Studying Microorganisms in the Living State

INTRODUCTION

Microorganisms include groups like bacteria, fungi, protozoa, some algae, and
parasites. These microorganisms manifest diverse characteristics. Bacteria, for example, are
differentiated by their shapes, staining properties, colony growths, and biochemical reactions
and motility, among others. Protozoans are classified according to whether they are motile or
not and based on the organelles that they use for locomotion. Bacteria move by means of
flagella whereas protozoans use either the flagella, cilia, or pseudopods for movement. True
motility should not be mistaken for Brownian movement which is the random movement
produced by currents in liquids causing organisms to vibrate and move.
Motile organisms found in pond water or hay infusions can be observed in their natural
living state using the wet mount method or the hanging drop preparation. The wet mount
method involves placing a drop of the specimen on a glass slide and covering it with a slip.
In the hanging drop preparation, a cover slip with a drop of the specimen is inverted over a
slide with a concavity at the center (hanging drop slide). Both preparations are then examined
under the microscope.

OBJECTIVES

At the end of the exercise, the students should be able to:

1. observe motile organisms from pond water or hay infusions;
2. identify some motile organisms from the samples; and
3. differentiate true motility from Brownian movement.

MATERIALS AND PREPARATIONS

Materials
Alcohol lamp Glass slides
Applicator sticks Compound microscopes Petroleum jelly
Cover slips Pond water/Hay infusion
Droppers Hanging drop slides Inoculating loop
Procedures

Wet mount preparation

1. On a clean slide, place a drop of pond water or hay infusion at the center.
2. Place a cover slip over the specimen and examine under the LPO.
3. Observe the microorganisms and take note of their movement.
Hanging drop method
1. Place a thin ring of petroleum jelly around the edge of the concavity of a hanging drop
slide.
2. Flame-sterilize an inoculating loop, then cool it for a few seconds. Use it to transfer two
loopfuls of pond water/hay infusion at the center of a cover slip.
3. Invert the hanging drop slide so that the petroleum-ringed concavity is directly over the
drop of water on the cover slip. Press lightly to seal the sample.
4. Re-invert quickly so that the water sample hangs suspended from the cover slip in the
concavity of the slide.
5. Do not allow the water sample to drop or touch the bottom of the concavity. If the water
drops, repeat the procedure.
6. Examine hanging drop slide under the LPO and observe the different motile organisms.
ILLUSTRATIONS

A. Draw the different organisms observed in the wet mount preparation.

 B. Draw the different organisms observed in the hanging drop method.

RESULTS AND DISCUSSION

 1. Based on your observations of microorganisms in their living state, which between the
wet mount method and the hanging drop method is better? Why?
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________
2. Aside from observing motility, what are the other uses of the wet mount and the hanging
drop preparations?
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________

EXPLORE AND RESEARCH

1. Prokaryotic Organisms like bacteria use the flagella for movement. What is the basic
structure of a flagellum?
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________

2. Motile bacteria have different numbers and arrangements of flagella. What are they? Cite
their difference.
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________

3. Give examples of motile bacteria and specify the number and arrangement of their
flagella.
 ________________________________________________________________________
________________________________________________________________________
______________________________________________________

4. How could you differentiate true motility from Brownian movement?

________________________________________________________________________
________________________________________________________________________
______________________________________________________

CONCLUSIONS

______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________.