by - OBJECTIVE:
The purpose of this document is to describe a procedure to be followed for Gram Staining of bacterial culture.
- SCOPE:
This SOP is applicable for the Gram Staining of bacterial culture at microbiology laboratory in conducted.
- RESPONSIBILITY:
Microbiologist
- ACCOUNTABILITY:
QC Head
- PROCEDURE:
- Out.
- He shall
- Flame the loop to red-hot.
- Take a drop of bacterial culture on the glass slide with the loop. Make a smear by spreading the drop evenly with the loop.
- Fix the smear by passing the slide over flame two to three times quickly.
- Place the glass slide with smear on stand and cover it completely with crystal violet solution for 1 minute.
- Wash with water and drain the excess water.
- Flood the smear with Gram’s iodine solution and keep for 1 minute.
- Wash with water and drain the excess water.
- Decolorize with ethyl alcohol (95%) for 30 seconds.
- Wash
with water and drain the excess water.
Cover the preparation with saffranine solution and keep for 30 seconds.
- Wash with water and drain the excess water.
- Dry the smear with tissue paper or allow it to air dry.
- Observe under microscope at 100X objective.
- TRAINING:
Trainer : Manager – Quality Control
Trainees : Staff of the microbiology departments
- DISTRIBUTION:
Master Copy : QA Department
Controlled Copy : Microbiology Department
Display Copy : Microbiology Department (If Required)
- ATTACHMENTS:
| Sr. No. | Title | Annexure No. |
| 01. | Gram Staining Record | F-SOP/MB/036/01-01 |
- REFERENCES:
In-House
- ABBREVIATIONS:
| Abbreviations | Extended Form |
| SOP | Standard Operating Procedure |
| NA | Not Applicable |
| MB | Microbiology |
| Sr. No. | Serial Number |
| QA | Quality Assurance |
| DEPT. | Department |
| IPA | Isopropyl Alcohol |
| % | Percentage |
