SOP on Motility test by Hanging drop Method


1.0 Objective:

To lay down the procedure for Motility test by Hanging drop Method.

2.0 Scope:

This SOP is applicable for Motility test by Hanging drop Method in pharmaceutical formulation plant.

3.0 Responsibility:

Microbiologist or above of Microbiology Laboratory: Preparation of SOP.

Head – Microbiology section / Nominee: Checking of the SOP.

4.0 Accountability:

Head – Quality Control / Nominee: Compliance of SOP.

5.0 Abbreviations:

SOP    : Standard operating procedure

No.      : Number

QC      : Quality Control

QA       : Quality Assurance

SP       : Specific

6.0 Procedure:

6.1 Make suspension of the microbial colony.

6.2 Apply grease or Vaseline around the depression of the hanging-drop slide.

6.3 Using the inoculating loop, aseptically transfer one drop of the culture to the center of a clean cover slip.

6.4 Invert the hanging-drop slide and center its well over the drop of culture. Press down on the edge of the cover slip so that grease makes a firm seal.

6.5 Quickly and carefully turn the slide right up so the hanging drop is suspended in the depression.

6.6 Do not let the drop fall or touch the bottom of the depression.

6.7 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.

6.8 You will see the edge as a bright wavy line against a gray background.

6.9 Switch to the High – dry objective (45 x) and refocus on the edge of the drop.

6.10 The bacteria within the drop will then be in approximate focus and detect them by their movement and by the contrast between them and the suspending media.

6.11 Differentiate true motility from Brownian movement carefully.

7.0 List of Annexure / Formats:

Not applicable

8.0 References (if any):

Not applicable

9.0 Distribution

9.1   Master Copy                                     : Documentation Cell (Quality Assurance)

9.2   Controlled Copies                            : Quality Assurance, Microbiology-QC

10.0 History

     Date    Revision Number                       Reason for Revision
                       New SOP



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