1.0 OBJECTIVE:
To lay down a standard procedure on motility test by hanging drop method.
2.0 SCOPE:
This Standard Operating Procedure is applicable for Microbiological Section of Quality Control department..
3.0 RESPONSIBILITY:
Microbiologist shall be responsible to follow the procedure mention in this SOP.
4.0 ACCOUNTABILITY:
Department Head & QA Head shall be accountable for effective implementation of this SOP.
5.0 ATTACHMENTS:
Nil
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 REFERENCES:
In-house
8.0 ABBREVIATIONS:
SOP: Standard operating procedure
°C : Degree Celsius
IPA : Isopropyl Alcohol
QA : Quality Assurance
QC : Quality Control
9.0 DISTRIBUTION LIST:
Quality Assurance
Quality Control
10.0 HISTORY OF REVISION:
| Version Number | Effective Date | Reason for Revision |
