DHANAJI NANA MAHAVIDYALAYA FAIZPUR DEAPRTMENT OF BOTANY PRACTICAL

DHANAJI NANA MAHAVIDYALAYA, FAIZPUR DEAPRTMENT OF BOTANY PRACTICAL COURSE BOT. 113 (BASED ON BOT. 111 & BOT. 112) Dr. N. A. Chaudhari

Practical No. 6 Gram Staining in bacteria

REQUIREMENTS CHEMICAL & STAINS Crystal violet Iodine Alcohol Safrannin

q. Materials ØMicroscope ØNeedle ØBlotting paper ØSlides ØSpirit Lamp ØDropper ØCurd

PROCEDURE Preparation of bacterial film Fixing the bacterial film Staining the bacterial film

Preparation of bacterial film 1. Take a clean slide and gently warm it over the spirit lamp. Let the slide cool. 2. Take a drop of curd with the help of dropper.

1. Place it over the one corner of the slide & spread evenly with another slide

Fixing the bacterial film 1. Pass the film quickly 3 -4 times over the flame of spirit lamp. 2. Let the slide cool so that the film firmly sticks to the slide.

Staining bacterial film 1. Stain the slide with crystal violet for about 1 minute.

2. Wash the slide with water 2 -3 time.

3. Put the slide in iodine for about 1 minute

4. Rinse the slide with water 2 -3 time.

5. Put the slide in 95% Acetone for about 30 seconds.

6. Wash the slide with water 2 -3 times.

7. Put the slide in counter stain safranin for about 30 seconds.

8. Wash the slide with water & dry with blotting paper.

9. Observe slide under microscope.

Observation Sr. no. Name of bacteria Morphology 1 Cocci Spherical or rounded 2 Bacilli Rod shaped Result Gram +ve /-ve

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