MICROBIOLOGY LABORATORY

standards to be met in the laboratory:

enter with lab apron

wash hands with soap and water

put on gloves and a mask

disinfect work areas

PRACTICE MATERIALS:

buccal biofilm sample

microscope

gram kit

gloves

face mask

lighter

PRACTICE: SPECIAL STAINS IN MICROBIOLOGY

PROCESS

TECHNIQUES OF GRAM STAINING.

1. i will Prepare a thin spread of the study material and let it air dry.

2. Fix the material to the slide so that it is not carried away during the staining process by
passing the slide 3 or 4 times through the flame of a Bunsen burner.

3. It will be placed the preparation on a staining support and cover the surface with Crystal
Violet solution for 1 minute.

4. Passing the minute, will wash with distilled water.

5. It will cover the preparation with Lugol Gram for 1 minute. Wash again with distilled water.

6. Hold the slide between thumb and forefinger and bathe the surface with a few drops of the
decolorizer: alcohol acetone until no more violet dye is dragged out. This usually takes about
30 seconds or less.

7. Will wash with distilled water and place the slide back on the holder. Cover the surface with
Safranina (counter color), for one minute. Wash off with water.

8. It will be placed the preparation on a staining rack in a vertical position, allowing excess
water to drain and the smear to dry.

9. Will be examined the smear microscope with and immersion objective (100X). Gram positive
bacteria appear deep blue or violet in color. Gram negative bacteria will appear red or pink in
color.