SOP of Gram Staining
Objective:
To lay down the procedure for Gram Staining This SOP is applicable for Gram Staining in Microbiology Laboratory of (Pharmaceutical Company Name).
- Responsibility
- Microbiologist or above of Microbiology Laboratory: Preparation of SOP.
- Head – Microbiology section / Nominee: Checking of the SOP.
- Accountability
- Head – Quality Control / Nominee: Compliance of SOP.
- Abbreviations and Definitions
SOP : Standard operating procedure
No. : Number
QC : Quality Control
QA : Quality Assurance
- Procedure
- Requirements: Crystal violet (primary stain), Gram’s iodine (mordant), 95% alcohol (decolorizing agent), Safranin (counter stain)
- Prepare smears of organisms by spreading cell mass from a colony or by taking a loopful from the broth as a thin film on the slide.
- Air dry, and fix the smear by passing the slide rapidly through the Bunsen flame for three times.
- Stain with crystal violet for about 1 min.
- Rinse with water.
- Cover the film with gram’s iodine and allow it to act for about 1 min.
- Rinse with water.
- Decolorize with 95% alcohol. For a thin smear, 10-20 seconds is long enough; after the proper time interval, alcohol drippings from the slide are no longer colored.
- Rinse with water. Counter stain with Safranin for about 20-30 seconds.
- Rinse with water and blot dry.
- Examine under the oil-immersion objective.
- Result: Gram-positive cells stain purple while. Gram-negative cells stain pink colour.
- Forms and Records (Annexures)
- Not applicable
- Distribution
- Master copy – Quality Assurance
- Controlled copies – Quality Assurance, Production, Quality Control, Stores, Engineering and Human Resource
- History:
| Date | Revision Number | Reason for Revision |
| 00 | NEW SOP |
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