gram stain Identification of microorganisms characteristics by gram staining method

Gram stain method for identification of gram positive gram negative bacteria.
PROCEDURES :
1 Pick up the microorganism to be identified and smear it on the clean slide with a
drop of sterile saline

2 Heat fix the smear on the slide by passing it through the flame 3 -4 times

3 Flood the slide with Grams crystal violet solution and keep it for i minutes

4 Wash with water and flood with Grams iodine and keep it for 1 minute

5 Wash with water and decolorise with Acetone alcohol solution till no further violet
colour discharges out

6 Wash with water and counter stain with 05 % saffaranin solution and keep it
for about 30 seconds

7 Wash the slide with water and air dry it

8 Observe the slide under oil immersion objective and observe for gram positive
and gram negative organisms Gram positive organisms will be violet in colour
and gram negative organism will be in pink colour

Standard operating procedure SOP for microbiology and aseptic techniques for manufacturing of sterile dosage forms injections

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Regulatory affairs in pharmaceutical industry

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