SOP of Motility test by Hanging drop Method
Objective:
To lay down the procedure for the Motility test by Hanging drop Method
Scope:
This SOP is applicable for the Motility test by Hanging drop Method.
Responsibility:
Microbiologist or above of Microbiology Laboratory: Preparation of SOP of Motility test.
Head – Microbiology section / Nominee: Checking the SOP of the Motility test.
Accountability:
Head – Quality Control / Nominee: Compliance of SOP of Motility test.
Definition : The motility test is utilized to determine whether an organism possesses the ability to move or not. Generally, motile organisms are bacilli, although there are a few motile cocci as well. This test is also employed to assist in distinguishing between different genera and species.
Procedure:
Make suspension of the microbial colony.
Apply grease or Vaseline around the depression of the hanging drop slide.
Using the inoculating loop, aseptically transfer one drop of the culture to the center of a clean cover slip.
Invert the hanging-drop slide and center it well over the drop of culture. Press down on the edge of the coverslip so that the grease makes a firm seal.
Quickly and carefully turn the slide right up so the hanging drop is suspended in the depression.
Do not let the drop fall or touch the bottom of the depression.
To examine the hanging drop, first locate its edge in the center of the microscopic field, with the low power objective (10 x) and markedly reduce the light.
You will see the edge as a bright wavy line against a gray background.
Switch to the High-dry objective (45 x) and refocus on the edge of the drop.
The bacteria within the drop will then be in approximate focus and detected by their movement and by the contrast between them and the suspending media.
Differentiate true motility from Brownian movement carefully.
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