FACULTY OF FOOD SCIENCE AND NUTRITION

NT20203 FOOD MICROBIOLOGY

TITLE:

PRACTICAL 3 - TECHNIQUES FOR THE OBSERVATION OF LIVE

ORGANISMS: HANGING DROP AND WET MOUNT

NAME: DESSY WEDANNIE BINTI WEI KUN

MATRIC NUMBER: BN20110128

LECTURER NAME: PROF. CHYE FOOK YEE & DR. KOH WEE YIN

PRACTICAL DATE: TUESDAY (10 MAY 2022)

DATE OF SUBMISSION: 17 MAY 2022

INTRODUCTION

Both living and non-living microorganisms can be recognized using microscopic preparations
(Erkmen, 2021). Direct examination of viable microorganism is very important in determining
and allows magnification of morphological appearance of cell such as size and shape, motility
and reaction toward various chemical agents and immunological antiserum. There are two
common techniques used in determining the viable microorganisms, which are hanging drop
and wet mount technique. These techniques are usually conducted without the addition of any
stains and both techniques are useful in indentifying whether or not the organism is motile.
Motility is the capability of an organism to move spontaneously.

MATERIALS AND METHODS

A. Wet mount technique

Materials

Clean slides, cover slips, nutrient broth culture (pond water sample), Pasteur pipette or dropper

Methods

1. 1 drop of culture is placed on the clean slides.

2. The cover slip is placed slowly on top of the slide with sample to avoid the presence of air
bubbles.

3. The prepared slide is observed under the microscope with low magnification power and then
with high magnification power.

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B. Hanging drop technique

Materials

Depression slides, cover slips, Vaseline, nutrient broth culture (pond water sample), Pasteur
pipette or dropper, toothpicks

Methods

1. Vaseline is applied to the surface edges of cover slip using a toothpick.

2. 1 drop of culture is placed in the center of the cover slip.

3. A depression slide is pressed slowly onto the cover slip, with the concave part over the drop.

4. The slide is then turned upside down with the cover slip on top of the slide.

5. The prepared slide is observed under the microscope with low magnification power and then
with high magnification power.

RESULTS

A. Wet mount technique

4x magnification 10x magnification

2
 40x magnification

Description: have motile organisms (rod-shaped bacteria), non motile organism (algae)

B. Hanging drop technique

4x magnification 10x magnification

3
 40x magnification

Descriptions: have motile organisms (rod-shaped bacteria), non motile organism (long-body
organism)

DISCUSSIONS

In this experiment, two methods have been applied to identify the type and motility
microorganisms in pond water sample which are wet mount and hanging drop method. Wet
mount method is very simple in which a drop of sample can be placed directly on the slide to be
observed under the microscope. Hanging drop technique is also often used for examining living
microorganisms. The procedure applies a depression slide with a circular concavity in the centre
and a drop of fluid containing the microorganisms is placed and hung from the cover slip
(Crawford, n.d.). Both methods does not need staining procedure because microbial cytoplasm
is usually transparent so they can be viewed with the light microscope (CliffNotes, 2022).
However, observation without staining can make it difficult to see the microorganisms so
reducing the light source intensity can help locating the microorganisms. Both techniques can
be applied to observe motility of the microorganisms. The differences between wet mount and
hanging drop technique is the rate of microorganisms movement are lower in wet mount

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because the slide tend to dry out quickly under the heat of the microscope light, so it is useful
for short-term observation only (Microbugz, n.d.). For hanging drop technique, the rate of
movement is constant as petroleum jelly (Vaseline) keeps the sample hydrated, so it can be
observed for longer period of time. In addition, the motility of microorganisms in wet mount are
limited as they are pressed in between the flat surface of the slide and cover slip meanwhile in
hanging drop method, the microorganisms can move freely because of the curvature on the
slide which provides more space for them to move (Amarillo et al., 2016). There are many
types of microorganisms found in aquatic ecosystems, including fresh water or pond. The main
aquatic groups that can be found include arthropods, bacteria, protozoa, hydras, fungi,
phytoplankton and algae.

CONCLUSIONS

In determining the microorganism, the two most common methods often used in the
microbiology laboratory are wet mount and hanging drop techniques. Hanging drop method is
considered better than wet mount in observing the motility of the microorganisms as the
sample are hanging on the cover slide and not pressed down like the wet mount method, so the
microorganisms can move freely for longer period of time.

REFERENCES

Amarillo, Bade R., Victoria A. & Celine. (2016, January 10). Activity 2 – Determination of
Bacterial Motility. https://www.slideshare.net/likewhatsoever/activity-2-determination-of-
bacterial-motility-56870098\

CliffsNotes. (2022). Staining Techniques. https://www.cliffsnotes.com/study-

guides/biology/microbiology/microscopy/staining-techniques

Crawford K. (n.d.). Microorganisms – a world in a hanging drop. Science & Plants for School.
https://www.saps.org.uk/secondary/teaching-resources/298-microorganisms-a-world-in-
a-hanging-drop

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Erkmen O. (2021). Wet-mount and hanging-drop techniques. Laboratory Practices in
Microbiology. https://www.researchgate.net/publication/349942220_Wet-
mount_and_hanging-drop_techniques

Microbugz. (n.d.). Wet Mount and Hanging Drop.

https://www.austincc.edu/microbugz/wet_mount_and_hanging_drop.php#:~:text=The
%20wet%20mount%20tend%20to,more%20reliable%20observation%20of%20motility.

Pond Informer. (2019, January 14). Types of Bacteria & Microorganisms In Pond Water (With
Pictures). https://pondinformer.com/types-of-pond-microorganisms/