SOP on Spore Staining

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1.0 Objective:

To lay down the procedure for spore staining.

2.0 Scope:

This SOP is applicable for spore staining in microbiology lab of pharmaceutical formulation plant.

3.0 Responsibility:

Microbiologist or above Microbiology laboratory: Preparation of the SOP.

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

4.0 Accountability:

Head – Quality Control/ Nominee: Compliance of SOPs.

5.0 Abbreviations:

SOP          : Standard operating procedure

No.            : Number

QC            : Quality Control

QA             : Quality Assurance

6.0 Procedure:

6.1 Prepare smear of the isolated pure culture.

6.2 Dry the smear in air and fix with heat.

6.3 Place the slide on a small beaker, containing boiling water, kept on the hot plate.

6.4 Cover the smear with a small piece of filter paper; keep the filter paper saturated with a 5% aqueous solution of malachite green for about 30 minutes.

6.5 Wash the slide gently with water.

6.6 Counter stain with safranin for 30 seconds.

6.7 Wash the slide with water and blot dry with whattman filter paper.

6.8 Observe the slide under microscope 100x using immersion oil.

6.9 The spores are stained green where as the vegetative cells are stained pink.

6.10 Observe the location of spore with in the cell. (E.g. Central spore, sub terminal spore, terminal spore with swollen sporangium.)

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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