Hanging Drop Technique
BIO112
Sanjay Kumar
([email protected])
The aim
To perform motility test of a bacteria in the laboratory,
by hanging drop preparation,
to find out whether it is motile or non-motile
Purpose
Motility is the ability of an organism to move independently
Based on motility, bacteria can be divided into two groups:
o Motile Bacteria:
A bacteria, which has the intrinsic ability of movement in the surrounding
medium, in which it remains suspended, is a motile bacteria.
o Non-motile Bacteria:
A bacteria, which does not have the intrinsic ability of movement in the
surrounding medium, in which it remains suspended, is a non-motile bacteria
Non-motile bacteria may show apparent motility, resulting from their Brownian
movement caused by the bombardment of the water molecules in the surrounding
medium, on the bacteria cells
How differ from WET MOUNT
In wet mount, though the shape and size of bacteria can be observed, motility may
be hampered, as the suspension is pressed between the slide and the cover slip
That is why; hanging drop preparation or motility test is performed for clear
observation of the motility of bacteria, besides their shape and size
It is useful in the identification of bacteria
Principle
A small drop of bacterial suspension is hung from the center of a cover slip
into the cavity of a cavity slide. The hanging drop is observed under a
microscope. If the bacteria are motile, its cells can be seen to have erratic
movement in the surrounding medium
if it is non-motile, its cells remain static in the medium without any
movement or may show Brownian movement resulting from the
bombardment by the water molecules in the medium, on the bacterial cells
Methods
1. Take a cavity slide and cover slip, wipe it with tissue paper to make it clean and grease free
(A cavity slide is a glass slide with a small round depression at the center, into which a
small drop of bacteria suspension can hang)
2. Apply Vaseline a ring or along the four corners of the cover slip
(To reduce the rate of evaporation and exclude the effect of air currents, to provide a seal
between the slide and cover slip)
3. Place a drop of sample on the cover slip with the help of Pasteur pipette
4. Gently cover the water drop with the cavity slide where concave shape falls on to the water
sample and press lightly
5. Observe the edge of the drop under the microscope under increasing magnification
The reasons for focusing the edge of the drop are :
(a) Better contrast is obtained due to difference in the refractive index of the drop and the cover slip
(b) As the drop hangs, it thins towards the edge, for which the edge contains less number of
bacteria to be observed clearly for motility
(a) Usually aerobic bacteria come towards the edge to get more oxygen for respiration, for
which they can be observed on the edge
Observations :
General shape of living bacteria, arrangement of bacterial cells
Motility
Motile or non-motile
Life in a pond
Filamentous Algae
Spirogyra is a genus of filamentous charophyte green algae
named for the helical or spiral arrangement
Zygnema is a genus of freshwater
filamentous thalloid alga
Free living algaes
Chlamydomonas is a genus of green algae
unicellular flagellates
Algae
Netrium (algae)
Protists
Perenema
Blepherisma
A protist is any eukaryotic organism
Euglena is a genus of single cell flagellate eukaryotics
